Background/Aim: Thespesia populnea is a plant known for its polyphenol and flavonoid content, which plays a crucial role in wound healing activity. Traditionally, extracts of various parts of T. populnea plant have been used to treat various skin diseases including wound healing. Therefore, we made a topical gel containing alcoholic extracts of the leaves of T. populnea for antimicrobial and wound healing activities. Materials and Methods: Extracts of T. populnea were successfully obtained by Soxhlet extraction. Phytochemical screenings were performed to estimate the presence of different metabolites. Combinations of carbopol and propylene glycol were optimized through 3² factorial design for desirable gel characteristics. Results and Discussions: Formulated gel was pale brown in color having viscosity of 90,300 cps with acceptable spreadability and extrudability index. Antimicrobial studies showed inhibition activity. The in vivo wound healing studies demonstrated comparable healing properties with respect to marketed Soframycin gel. Furthermore, histopathological studies on Wistar rats also confirmed the same. Conclusion: It can be concluded that the formulated gel is beneficial in topical applications, which can be a beacon for new skin regeneration and wound healing therapies that focuses on herbal remediation.

Background and Objectives: Oxalate is a toxic metabolite, which is predominantly excreted through the kidneys. Hyperoxaluria is a clinical condition characterized by the occurrence of excessive amounts of oxalate in the urine. Hyperoxaluria affects most of the patients with kidney stone. In this study, we aimed to reveal the therapeutic actions of escin against the hyperoxaluria-induced nephropathy in rats through the suppression of inflammation and oxidative stress. Materials and Methods: Wistar rats were induced with hyperoxaluria via administration of 0.4% ethylene glycol and 1% ammonium chloride through drinking water and treated with 50 mg/kg of escin for 28 days. The bodyweight, water intake, and urinary output was monitored and tabulated. The renal markers such as urinary kidney injury molecule-1 (KIM-1), N-acetyl-β-D-glucosaminidase (NAG), and lactate dehydrogenase (LDH) were examined using assay kits. The pro-inflammatory cytokines such as IL-1 β, IL-6, and monocyte chemoattractant protein-1 (MCP-1) were quantified using kits. The MDA and antioxidant enzymes such as superoxide/dismutase (SOD), glutathione peroxidase (GPx), and glutathione reductase (GR) were investigated using kits. The mRNA expression of CCAAT/enhancer-binding protein homologous protein and GRP78 was studied by reverse transcription–polymerase chain reaction analysis. Results: Escin remarkably improved the bodyweight and decreased the renal weight, water uptake, and urinary output in Wistar rats. The status of urinary oxalate, LDH, NAG, and KIM-1 was appreciably suppressed by the escin treatment. Escin also reduced the levels of interleukin (IL)-1 β, IL-6, MCP-1, and lipid peroxides and increased the activity of antioxidant enzymes such as SOD, GPx, and GR. It downregulated the expression of CHOP and GRP78. Furthermore, histological examination of escin-treated hyperoxaluric animals revealed improved kidney structures. Conclusion: The results of this study revealed that escin shows potent therapeutic actions against hyperoxaluria-induced nephropathy in rats.

Background: Piper retrofractum (PR) is traditional medicine in the South-East country. Objectives: This study examined the PR effects on anticancer and anti-migratory activity in human breast cancer cells. Materials and Methods: Determination of cancer cell death by the sulforhodamine B, colony formation, flow cytometry methods. Mechanism-induced cells death was assessed by intracellular reactive oxygen species generation, caspase 3 activity, mitochondrial function, gene expression, and protein expression. Results: The ethanolic extract of PR had higher effects against MCF-7 cells than DW extract expressing the high levels of piperine. The extracts stimulated cell death in MCF-7 cells and induced apoptosis with increasing caspase 3 activity, ROS generation, and decreasing mitochondria function. Moreover, the extract also inhibited gene-related cell proliferation, cdk2, cdk4, and ckd6, and altered protein-related cell growth, cyclin D1 reduction and p21 induction. Finally, the extract caused inhibition of cells migration by reducing MMP 9 levels. The extracts of PR stimulated MCF-7 cells death in association with increasing apoptosis and inhibiting cancer cells migration. The induction of cancer cell death may be through modulating mitochondrial function. Conclusion: These extracts may be a novel strategy to improve the efficacy of chemotherapy to treat MCF-7.

Background: Alzheimer disease (AD) is a common form of dementia and is described by memory loss and behavioral disorder. The prevalence of AD is increasing rapidly each year worldwide. Objectives: In this study, we aimed to discover the therapeutic properties of esculetin against D-galactose (D-gal)-induced AD in an animal model. Materials and Methods: AD was initiated in rats by administering 150 mg/kg of D-gal via subcutaneous route for 6 weeks and supplemented with 10, 20, and 30 mg/kg of esculetin, respectively. Subsequently, memory and learning of the rats were investigated using the Morris water maze (MWM). The organ index of the liver, spleen, thymus, and kidneys was assessed. The enzyme activities of superoxide dismutase (SOD), catalase (CAT), GSH-Px, and heme oxygenase-1 (HO-1) and the levels of advanced glycation end products (AGEs), 8-iso-prostaglandin F (8-iso-PGF), and 8-hydroxy-2-deoxyguanosine (8-OHdG) were assessed using commercially available kits. The level of acetylcholine (Ach) and the activity of acetylcholinesterase (AChE) was also assessed using kits. The brain tissue samples were assessed microscopically. Results: According to the results, esculetin significantly improved the bodyweight and organ index in AD animals. It significantly modulated the spatial learning and memory and improved the activities of CAT, SOD, GSH-Px, and HO-1. It significantly reduced the contents of AGEs, 8-iso-PGF, and 8-OHdG and inflammatory markers. Furthermore, esculetin increased the level of ACh and the reduced activity of AChE. Histological analysis of the brain tissue revealed that esculetin attenuated the D-gal-induced histological changes in the brain of AD rats. Conclusion: The findings of this study reveal that esculetin can ameliorate inflammation and oxidative damage in D-gal-induced AD rats. It can be further explored as a therapeutic agent to treat AD.

Background: Tinospora cordifolia (Willd.) Miers (T. cordifolia) is a well-known Indian medicinal plant containing several nonpolar and polar constituents that play an important role to mitigate various ailments, such as diabetes, urinary disorders, and hepatoprotective. Due to the lack of evidence on phytopharmacological relevance to the unpredicted nonpolar matrix of T. cordifolia, the present study aimed to evaluate the metabolomic pattern of different fractions obtained from aqueous extract of T. cordifolia, which has been recommended in AYUSH for various ailments including kidney disorders. Materials and Methods: High-performance thin-layer chromatography and gas chromatography–mass spectrometry (GC–MS) analyses were performed on aqueous extracts and hexane, dichloromethane, and methanolic fraction of T. cordifolia aqueous extract to evaluate fingerprinting and metabolomic profile. Principal components and pharmacokinetic analysis were performed using XLSTAT and in-silico SwissADME tool to determine metabolite variability and pharmacokinetic relationship based on lipophilicity and drug-likeness. Further, network pharmacology analysis was performed to determine the exact biomolecular relationship of T. cordifolia in alleviating kidney disease. Results: The GC–MS metabolomics results showed several metabolites in different fractions with high variability of phytoconstituents in the methanolic fraction. In pharmacokinetics, each metabolite exhibited a direct correlation between drug lipophilicity and permeability. Network pharmacological suggested five fatty acids, which significantly interacted with the genes such as AGTR1, ATG, RELA, NOS3, NOS2, REN, INS, IL6, TNF, MAPK1, and CASP3, which could potentially regulate various pathophysiological conditions, such as hypertension, insulin resistance, oxidative and inflammatory stress, and electrolyte homeostasis, thereby strengthening the normal function of the kidney. Conclusion: The study showed that six metabolites of T. cordifolia play a multimechanistic role in alleviating kidney disease.

Background: The genus Gloriosa is commercially valued due to its colchicine metabolite, which is clinically used in gout and as an antimitotic agent. Objectives: The study was a comparative pharmacognostical evaluation and simultaneous high performance thin layer chromatography (HPTLC) quantification of bioactive alkaloids in G. superba, G. lutea and G. rothschildiana. In vitro anti-gout activity was also established. Materials and Methods: Pharmacognostical studies and metabolic variations were analyzed per the Ayurvedic Pharmacopoeia of India and validated HPTLC method. Inhibition of protein denaturation, hydroxyl radical scavenging and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay evaluated the anti-gout activity. Results: The morpho-anatomical studies suggest that the target species are similar with no characteristic difference, apart from the flower colour. The pharmacognostic standards were also established as per the Ayurvedic Pharmacopoeia of India to ensure the quality of raw material. Quantification of colchicine and gloriosine content through validated HPTLC method reveals significant variation, ranging from 0.046% to 0.860%, and from 0.040% to 0.198% on dry wt. basis. The maximum content of both the targeted metabolites was in G. superba, followed by G. lutea and G. rothschildiana. The highest in vitro anti-gout and radical scavenging activity among the three species was in G. superba. Conclusion: The pharmacognostic standards of three Gloriosa species were established, and this opens avenues for chemotaxonomic studies on G. lutea and G. rothschildiana from different phytogeographical zones of India for the identification of their elite germplasm. The study also led to the identification of alternate species of G. superba which can be explored commercially to meet the industrial demand for colchicine.

Background: Flavonoids constitute one of the best-characterized groups of plant secondary metabolites with enormous pharmaceutical potential. A flavone type of plant flavonoid, cirsilineol, has been reported to exhibit proapoptotic effects against malignant human cells. Objectives: The present study was designed to investigate the antiproliferative effects of cirsilineol against human gastric cancer cells. Materials and Methods: Cell viability was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and colony formation assays. Apoptosis was detected by acridine orange/ethidium bromide (AO/EB) and annexin V/propidium iodide (PI) assay. Protein expression was examined by western blotting analysis. Results: The results showed cirsilineol inhibits the proliferation of human gastric cancer cells. The IC₅₀ of cirsilineol against human gastric cancer cells (BGC-823, SGC-7901, and MGC-803) ranged from 8 to 10 μM. Nonetheless, cirsilineol exhibited comparatively lower antiproliferative effects against normal GES-1 cells. The IC₅₀ of cirsilineol against normal GES-1 cells was found to be 120 μM. Colony formation assay showed that cirsilineol suppressed the colony formation of BGC-823 and MGC-803 cells in a dose-dependent manner. Acridine orange and ethidium bromide (AO/EB) staining showed that cirsilineol induced apoptosis in BGC-823 and MGC-803 cells. The percentage of apoptosis increased from 7.4% in control to 40.5% in BGC-823 cells and from 6.56% in control to 33.53% in MGC-803 cells at 8 μM cirsilineol. Western blotting showed cirsilineol caused an increase in Bax and cleaved caspase-3 and a decrease in Bcl-2 expression in both BGC-823 and MGC-803 cells. Conclusion: Together, the results are indicative of the proapoptotic and antitumor potential of cirsilineol against gastric cancer cells, suggestive of its possible therapeutic significance in future.

Background: Rheumatoid arthritis (RA) is an autoimmune disease that seriously affects the patient's quality of life. Total glucosides of peony (TGP) have long been used to treat RA in China despite the need for long-term administration. The main active ingredients in TGP are paeoniflorin (PF) and albiflorin (AF). This study aimed to clarify the effective mechanism of TGP only after long-term administration, and the pharmacokinetics of PF and AF following single and chronic oral TGP administration in collagen-induced arthritic (CIA) and normal rats. Materials and Methods: Comparative pharmacokinetic studies of TGP were conducted in male Sprague Dawley rats. Plasma concentrations of PF and AF were determined with ultra-high performance liquid chromatography–tandem mass spectrometry (UPLC-MS/MS). DAS software was used to estimate pharmacokinetic parameters. Results: Bimodal phenomenon was observed. After a single TGP dose, the absorption of AF and PF were both lower in CIA rats compared to control rats. Compared with single-dose groups, the absorption of AF and PF in CIA rats increased after long-term administration of TGP. No significant differences were seen between the groups after chronic administration in low-dose groups in normal rats. Conclusion: We found that after long-term administration of TGP, the absorption of PF and AF are promoted in CIA rats. These results, combined with the existing literature, may help substantiate the TGP-related changes in gut microbiota.

Background: Keloid is a fibrotic disease characterized by hyperproliferative fibroblasts. Notoginsenoside R1 (NGR1) possesses inhibitory roles on cell proliferation. Thus, the research sought to assess the mechanism of action of NGR1 against keloid. Materials and Methods: Cell viability of normal and keloid fibroblasts pretreated with different NGR1 concentrations was determined by Cell Counting Kit-8 assay. Cell cycle, apoptosis rate, and tube length were detected using flow cytometry and tube formation assay. Vascular endothelial growth factor (VEGF) expression was measured by quantitative real-time polymerase chain reaction and western blot. To verify the reversal effect of VEGF on NGR1, KEL FIB cells were transfected with pcDNA3-VEGF plasmids following treatment with 40 μM NGR1; subsequently, the above indicators were determined again. Results: NGR1 decreased cell viability, and 20, 30, and 40 μM NGR1 concentrations were selected for the next investigation. After KEL FIB was treated with NGR1, the apoptosis rate was increased, cell cycle was arrested, and tube formation was suppressed in a dose-dependent manner. The expression of VEGF was also suppressed. In further experiments, cell cycle and tube formation were promoted and apoptosis rate was decreased in NGR1-treated cells when VEGF was overexpressed. Conclusion: NGR1 may play as an inhibitor of endogenous VEGF, and NGR1 exerts its inhibitory effects on keloid fibroblasts by downregulating VEGF.

Background: Oral cancer is one of the cancers that pose a great threat to life globally. Though a number of chemotherapeutic strategies have been followed to treat oral cancer, a number of adverse effects limit the usage of chemotherapeutic agents. Purpose: In the current study, we evaluated the anticancer potential of brucine. Materials and Methods: KB cell line was treated with various concentrations of brucine (10–100 μg/ml) and the cytotoxicity and cell viability were measured. Acridine orange/ethidium bromide (AO/EB) staining was performed to assess apoptosis in cells. To evaluate the mechanism of brucine-mediated apoptosis, gene expression studies were carried out (Bax and cMyc). Results: Brucine caused cytotoxicity and reduced cell viability as assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and trypan blue exclusion assay, respectively. The IC₅₀ value of brucine was calculated to be 30 μg/ml. Further experiments were performed according to the IC₅₀ value. Brucine-treated cells were rounded up and detached from the surface. AO/EB staining revealed apoptotic mode of cell death. Further, our experiments revealed elevated levels of reactive oxygen species (ROS) within the cells, which might have caused apoptosis. A dose-dependent rise in lipid peroxidation was evident from thiobarbituric acid reactive substance (TBARS) assay, which could be due to the oxidative stress caused by brucine. Nitric oxide levels were impaired by brucine, which correlated well with apoptosis. On the other hand, catalase activity was decreased by brucine in a dose-dependent manner, implying the reason for oxidative stress. Polymerase chain reaction (PCR) analysis revealed increased expression of Bax and impaired expression of the oncogene cMyc. Conclusion: Brucine triggers oxidative stress, which leads to lipid peroxidation and induces the expression of the proapoptotic gene Bax, thereby causing apoptosis. In parallel, brucine-mediated suppression of cMyc leads to impairment of metastasis.

Background: Nociception is an unpleasant experience that has a negative effect on both the physiological and psychological status of individuals. Factors including temperature, and physical damage originating from mechanical, chemical, or even biological stimuli are likely to induce pain. Currently, a limited choice of antinociceptive medications is available for the management of pain. Purpose: Pharmacological activities of various plant extracts and derivatives are being actively explored for discovering novel plant-based antinociceptive agents. In the current work, we explored Adhatoda vasica for its antinociceptive activities. Materials and Methods: In the current experiment, we used zebrafish (Danio rerio) as a model organism, and to induce pain, formalin was administered. We used extracts from A. vasica at concentrations of 5, 10, 15, and 20 mg/ml for determination of the antinociceptive effect. To evaluate the antinociceptive activity of the A. vasica extract, animal behavior, reactive oxygen species (ROS) quantification, nitric oxide (NO) estimation, and reduced glutathione (GSH) estimation were performed. For behavioral analysis, the animal movement was recorded as a movie and analyzed using Behavioral Observation of Research Interactive Software (BORIS). Results: The behavior of fish was recorded by BORIS. Fishes treated with formalin preferred to stay in darkness. In addition, they also maintained their position at the bottom of the water tank. Upon treatment with the plant extract, the fish moved to light and the middle layer of the tank. The plant extract ameliorated the pain in fish, as evidenced by the swimming pattern. Further, the plant extract reduced the ROS and NO. Conclusion: The A. vasica plant extract reduced inflammation, as evidenced by a reduction in the inflammatory markers. The antinociceptive activity was further indicated by restoration of the color change induced by formalin. In conclusion, the current study emphasizes the significance of A. vasica plant extract in relieving pain.

Background: Sinomenine (SIN) plays a role in regulating intestinal immune inflammation, but its effect on the intestinal immune response is unclear. Objective: To investigate the potential mechanism of SIN in protecting intestinal immunity. Materials and Methods: The mechanism by which SIN regulates intestinal immunity was detected in RAW264.7 cells using enzyme-linked immunosorbent assay, real-time quantitative polymerase chain reaction, Western blotting, and immunohistochemical staining. Results: Compared with the control group, the LPS group has higher cell viability and inflammatory cytokines (interleukine-1beta [IL-1β], tumor necrosis factor α[TNF-α], IL-6, IL-17A, and IL-23), chemokine, and metalloproteinase levels. SIN significantly suppressed these increases. By contrast, aromatic hydrocarbon receptor (AhR) and IL-10 levels were lower in the LPS group compared with the control group, and SIN treatment prevented increased these levels. Conclusion: SIN can activate the innate immune function of the intestinal tract by affecting the IL-23/IL-17 axis through the AhR.

Background: Dolichandrone serrulata flower has been known for its antioxidation and anti-inflammation. Its ability to induce cell death and reduce migration on cancer cell lines has been previously demonstrated. Aim: The present study aimed to investigate the molecular mechanisms involved in the antimigration effect of D. serrulata flower ethanolic extracts of HeLa cell line. Materials and Methods: HeLa cell line viability was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The antimigration of the extract was investigated using a wound healing assay. Its possible molecular mechanism for the migration of HeLa cell line was investigated using a protein array assay. Results: D. serrulata extract caused cell death with CC₅₀ of 232.10 ± 17.16 μg/mL. The result from the wound healing assay demonstrated that D. serrulata extracts at concentrations of 62.5, 125, and 250 μg/mL caused the reduction of the narrowing in the wound site, corresponding to the progress over time. To eradicate the possibility of the cytotoxic effect of the herb, D. serrulata at a concentration of 125 μg/mL was further observed for protein expression using a protein array assay. The result showed the reduction of galactin-3, vimentin, and fibroblast growth factor-2 expression compared with the control group. Conclusion: Antimigratory effect of D. serrulata flower ethanolic extracts might be attributable to the reduction of vimentin and fibroblast growth factor-2 gene expression, but not galactin-3-induced apoptosis. Further study is needed to confirm and investigate a more in-depth pathway on its antimigration effect in the cervical cancer cell line.

Background: Bovine viral diarrhea virus (BVDV) has a serious impact on the global livestock industry; however, there are no specific therapeutic drugs for BVDV, so the development of anti-BVDV drugs is a research priority. Objectives: To investigate whether baicalin and its ester derivatives are active against BVDV non-structural protein 5B (NS5B). Materials and Methods: We modified the sugar chain part of the structure of baicalin by esterifying the carboxyl group at the 6-position of 7-β-d-glucuronide and by introducing alkyl groups of different lengths. The binding and in vitro activity of the baicalin ester derivatives with BVDV NS5B polymerase was determined using molecular docking, molecular dynamics, Cell Counting Kit-8 (CCK-8) assays, and real-time RT-PCR. Results: The following six baicalin ester derivatives were obtained: baicalin methyl ester, baicalin ethyl ester, baicalin propyl ester, baicalin butyl ester, baicalin hexyl ester, and baicalin heptyl ester. Molecular docking, molecular dynamics, CCK-8 assays, and real-time RT-PCR showed that baicalin and its derivatives could bind to BVDV NS5B polymerase, with baicalin ethyl ester showing the best binding ability and antiviral activity. Conclusion: Baicalin and its ester derivatives exert an inhibitory effect on BVDV by targeting the BVDV NS5B polymerase, and this effect shows a structure–activity relationship. The results provide an important theoretical basis for the further development of anti-BVDV drugs.

Background: Oroxylum indicum (L.) Kurz is an edible plant that is widely consumed in several Asian countries. In our previous study, it was demonstrated that Oroxylum indicum (L.) Kurz seed (OIS) extract attenuated oxidative stress and proinflammatory cytokines in activated microglia. However, the molecular mechanism remains unknown. Objectives: To confirm the antiinflammatory and antioxidative effects of OIS extract on and molecular mechanisms of OIS in lipopolysaccharide (LPS)-induced microglial activation. Materials and Methods: Nitrite and Chloro methyl derivative of 2', 7'-dichlorodihydrofluoresceindiacetate (CM-H₂DCFDA) assays were performed to examine the effect of OIS extract on LPS-induced nitric oxide (NO) and reactive oxygen species (ROS) production, respectively. Immunofluorescence and ELISA were performed to detect the levels of nuclear factor-κB (NF-κB) in the nucleus. Western blot analysis was performed to detect the phosphorylated (p-) form of Akt and extracellular-signal-regulated kinase 1/2 (ERK1/2). Results: OIS extract exhibited a significant and dose-dependent suppressive effect on LPS-induced NO and ROS production. Mechanistically, OIS extract attenuated the p-Akt and p-ERK1/2 signaling pathways, thereby inhibiting the LPS-induced NF-κB nuclear translocation. Conclusion: OIS extract exerts anti-oxidative effects by suppressing ROS production, and anti-inflammatory effects by inhibiting NO in activated BV2 microglial cells. Mechanistically, OIS extract could inhibit the Akt/ERK1/2-mediated NF-κB pathway. OIS extract may act as a promising functional food for the alleviation of neuroinflammation in neurodegenerative diseases.

Background: Luteolin-7-O-glucoside (Lut-7-G) is an effective compound found in plants, such as Patrinia and honeysuckle. It has anti-inflammatory as well as antioxidant properties; however, its anti-inflammatory effect on ulcerative colitis (UC) is hardly understood. Objectives: We evaluated the effects of Lut-7-G in dextran sodium sulfate (DSS)-induced UC in mice models, and then explore the underlying mechanism by studying the JAK1/STAT6/SOCS1 signaling pathway. Materials and Methods: Induction of acute colitis in mice was achieved by feeding 2.5% DSS for 7 days. Bodyweight loss, colon length, disease activity index (DAI) score, and spleen index were determined and hematoxylin-eosin and Periodic Acid-Schiff staining were performed to study the pathological changes in mouse colons. The inflammatory factor levels were determined by ELISA, JAK1, STAT6, and SOCS1 expression in colon tissues by RT-qPCR, and signaling pathway proteins by Western blotting. Results: It was found that treatment with Lut-7-G reduced the effects of colon shortening and weight loss, DAI score, spleen index, as well as colon inflammation. In addition, it significantly decreased DSS-induced overexpression of IL-6, IL-1β, IL-18, as well as TNF-α, and considerably reduced mRNA expression of JAK1 and STAT6 but upregulated the SOCS1 expression. Furthermore, Lut-7-G treatment dose-dependently decreased JAK1 and STAT6 protein expression, and only DSS + Lut-7-G (100 mg/kg) could downregulate p-JAK1 and p-STAT6 expression and upregulate SOCS1 protein expression. Moreover, Lut-7-G (100 mg/kg) was as effective as mesalazine. Conclusion: Lut-7-G may regulate the secretion of inflammatory factors and inhibit inflammatory responses through the JAK1/STAT6/SOCS1 pathway, as a determinant in the treatment of UC.

Background: This investigation was planned to evaluate the anti-cancer effects of royal jelly (RJ) obtained from Apis mellifera compared with doxorubicin as an anthracycline with potent anti-cancer activity and its cellular mechanisms against the human hepatoma cell line HepG2. Materials and Methods: The cytotoxic effects of various concentrations of RJ on the HepG2 cell viability by the 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide assay were studied. For the primary and late apoptosis in HepG2 cells exposed with RJ, we used the Annexin-V (AV) assay using cytometry analysis using the commercial kit as explained by the manufacturer's guidelines. Real-time polymerase chain reaction assessed the gene expression of miRNA-34a (miR-34a), caspase-3, Bcl-2, and Bax. We also used the western blot to evaluate the protein expression levels of poly (ADP-ribose) polymerases (PARP), Caspase-3, Caspase-9, Bcl-2, and Bax. Results: The IC₅₀ value of RJ was found 1.13 mg/mL for HepG2 cells. RJ revealed no cytotoxicity on normal THLE-3 cells with IC₅₀ >2 mg/ml. RJ at the concentrations of ½ IC₅₀ significantly increased (p < 0.05) apoptotic and necrotic cells from 0.96% to 28.3% and 9.3%, respectively. RJ at the concentration of IC₅₀ significantly increased (p < 0.05) apoptotic and necrotic cells from 0.96% to 39.2% and 14.12%, respectively. The expression of miR-34a, Caspase-3, and the Bax gene was considerably (p < 0.001) up-regulated as they are dose-dependent, whereas the expression level Bcl-2 was considerably (p < 0.05) declined in the HepG2 cells exposed with RJ. Treatment of HepG2 cells treated with RJ triggered a significant inhibition of Bcl-2 protein, whereas a significant rise in PARP, Caspase-3, Caspase-9, and Bax expression was observed. Conclusion: Our results showed the promising anti-cancer effects of RJ against HepG2 cells, whereas the induction of apoptosis by various pathways is considered the main mechanism underlying the cytotoxic effect of RJ against HepG2 cells. The present study's findings propose that RJ can be a candidate agent for treating human HCC.

Background: Hepatolithiasis (HL) is one of the common diseases in hepatobiliary surgery, and bile duct fibrosis is the key step in the pathogenesis of HL. Salvia miltiorrhiza (S. miltiorrhiza) has an inhibitory effect on bile duct fibrosis, but its molecular mechanism remains unclear. Objectives: Our research aimed to clarify the molecular mechanism of S. miltiorrhiza on HL. Materials and Methods: The rats were fed with salvianolic acid B and salvianolic acid B-free lithogenic diet (LD) for 4 weeks to observe the degree of bile duct fibrosis. The expression levels of TNF, IL1B, IL6, and MRP2 in HIBEC stimulated by taurodeoxycholic acid (TDCA) were measured by RT-qPCR and ELISA. E-cadherin and Vimentin expression in HIBEC was detected using immunofluorescence and Western blot. Cell proliferation was detected by CCK8. Results: The fibrosis of the bile duct was the key step of HL. The NF-κB pathway was activated and MRP2 was expressed low in intrahepatic biliary epithelial cells surrounding bile duct stones. Through experiments, salvianolic acid B (Sal B) delays HL via the NF-κB/MRP2 axis. Conclusion: In this research, we confirmed that salvianolic acid B inhibited the HL via the NF-κB/MRP2 axis.

Background: Numerous reports on the side effects of anti-cancer drugs have encouraged scientists to search for alternative anti-cancer agents with higher efficacy and fewer side effects. The current investigation was intended to study the anti-tumor efficacy of Terfezia claveryi Chatin methanolic extract (TCME) on mice with Ehrlich solid tumors (EST). Materials and Methods: EST mice received TCME at doses of 50 and 100 mg/kg orally once a day for two weeks. To study the anti-tumor effects, the rate of tumor growth, weight of the body, the serum level of some tumor markers, enzymes related to liver and kidney function, oxidant and antioxidant enzymes, tumor necrosis factor alpha (TNF-α) level, and some genes related to apoptosis were investigated. Results: The results revealed that the growth rate of the tumor, tumor markers, oxidative markers, enzymes related to liver and kidney function, level of TNF-α, and Bcl-2 gene expression were considerably declined in the EST mice receiving the TCME, whereas the enzyme levels related to antioxidant activity as well as Bax and caspase-3 gene expression were considerably elevated (P < 0.001). Conclusion: We found that T. claveryi methanolic extract has relevant anti-tumor efficacy on mice with EST and might be considered as a substitute anti-cancer compound; however, more studies especially in vulnerable humans are mandatory to approve these outcomes.

Background/Context: The fibrous roots of Rhizoma Anemarrhena (FRRA) are produced in the primary processing of Rhizoma Anemarrhena (RA). The lifespan-prolonging effect and the bioactive compounds of FRRA were rarely reported. Objectives: To study the lifespan-prolonging effects and reveal the potential bioactive components of FRRA. Materials and Methods: Caenorhabditis elegans were incubated with FRRA extracts (0.25, 0.5, 1.0 mg/mL). The number of live worms was recorded till all worms were dead. The components in FRRA or in vivo in Caenorhabditis elegans were detected and identified via UHPLC-Q-TOF-MS and Unify Software. The lifespan-prolonging effect of the components detected in vivo in Caenorhabditis elegans was verified. The contents of the components detected in vivo in Caenorhabditis elegans in FRRA and RA were determined and compared using UHPLC-MS. Results: FRRA extracts (0.5 mg/mL) significantly (**P < 0.01) extended the lifespan of the Caenorhabditis elegans (mean value from 15.07 to 19.10 days). A total of 28 components were detected and identified in FRRA, and 4 components were first screened in FRRA. A total of 5 components including neomangiferin, mangiferin, isomangiferin, timosaponin B-II and timosaponin A-III were detected in vivo in Caenorhabditis elegans and the effective concentration for the 5 components were 100, 100, 100, 120 and 120 μM, respectively. The average contents of neomangiferin, mangiferin, isomangiferin, timosaponin B-II and timosaponin A-III in FRRA were 0.80, 1.44, 0.50, 3.62 and 0.72%, respectively. Conclusion: FRRA is rich in lifespan-prolonging compounds and could be used as a potential drug resource.

Background: Despite tremendous efforts that have been made, cancer is still the leading cause of death all over the world. Chemotherapy, considered a routine method, always faces severe side effects and drug resistance. Cordyceps militaris (C. militaris) is a kind of folk tonic food and traditional Chinese medicine and was reported to have anticancer capacity. Since it is difficult to cure cancer via chemotherapy, preventing or inhibiting malignant cells by diet behavior seems useful and attractive. Objectives: In this study, we aim to assess the anticancer capacity of C. militaris polysaccharide (CMPs) and explore its anticancer mechanism. Materials and Methods: Polysaccharide was extracted by water from C. militaris. Its total sugar, protein percentage, and monosaccharide composition were measured via the phenol-sulfuric acid method, Bradford kit, and GC assay, respectively. The anti-proliferation effect of the CMPs was screened against several cancer cell lines by CCK8. Its anticancer effect was further studied by cell morphology, live/dead cell staining, and cell apoptosis study. Cellular reactive oxygen species (ROS) evaluation and western blots assay were conducted to explore its anticancer mechanism. Results: According to our data, the CMPs could effectively inhibit the proliferation of cancer cells, with IC₅₀ values ranging from 437.8 μg/mL to 545.1 μg/mL. Administration of CMPs could cause morphological change among cells and induce cell apoptosis. The study mechanism revealed that the CMPs exerted an anticancer effect via increasing the cellular ROS level and activating the endogenous apoptosis pathway. Conclusion: The CMPs can effectively inhibit cancer cells via arousing cellular ROS and activating the endogenous apoptosis pathway.

Background: Benzyl isothiocyanate (BITC) is a natural compound found in numerous cruciferous vegetables, and research has indicated that it has diverse biological activities. Isothiocyanate and its derivatives are the major anticancer natural compounds in cruciferous vegetables; these compounds help inhibit tumor cell proliferation through various mechanisms such as promoting tumor cell apoptosis, prompting cycle arrest, and increasing the generation of reactive oxygen species (ROS). Objectives: In human pancreatic cancer, gemcitabine is the first-line treatment; however, pancreatic cancer cells readily develop resistance to gemcitabine. Studies have demonstrated that natural products can promote the effect of gemcitabine and enhance the apoptosis process; however, the relevant mechanism and potential of BITC in human pancreatic cancer cells with gemcitabine resistance, namely, MIA PaCa-2/GemR cells (MIA RG100), are unclear. Materials and Methods: To elucidate the extent to which BITC induces apoptosis, we investigated the time and dose-dependent cell viability of PaCa-2/GemR cells under treatment with BITC. Results: Following BITC treatment, the PaCa-2/GemR cells exhibited DNA condensation, as indicated by transferase-mediated d-UTP nick end labeling (TUNEL) stain, with a corresponding increase in ROS production in mitochondria. Moreover, colorimetric assay analyses revealed that BITC increased caspase-9 and caspase-3 activities in PaCa-2/GemR cells. Our results indicate that BITC induces apoptotic cell death in PaCa-2/GemR cells through a mitochondrial-dependent signaling pathway.

Background: Curcumin possesses multifunctional pharmacological properties, including antioxidant, anti-inflammatory, and antidiabetic properties. We investigated whether curcumin can improve pathological changes associated with Alzheimer's disease (AD), including amyloid β (Aβ), protein kinase B (PKB, also termed Akt), and glycogen synthase kinase 3β (GSK3β) levels and expression. Materials and Methods: Alzheimer's transgenic APP_SWE/PS1Δ_E9 mice and wild-type mice were treated with curcumin by intragastric administration for 2 weeks at 2 and 5 months of age, respectively. Aβ plaques and contents in the brain and retina were measured by immunohistochemistry and enzyme-linked immune sorbent assay, respectively, while the expression of Akt and GSK3β was tested by RNA isolation and quantitative real-time polymerase chain reaction. Blood of patients with AD and age-matched healthy controls was used to determine the contents of Aβ, Akt, and GSK3β. Results: Curcumin treatment decreased Aβ accumulation in the early stages of AD at 5 months (P < 0.001). It also improved AD-associated pathological changes, including upregulation of Akt (P < 0.01) and downregulation of GSK3β (P < 0.01). In addition to AD-associated changes, the proinflammatory cytokine interleukin (IL)-1β was significantly decreased with curcumin treatment (P < 0.05). Conclusion: In the early stage of AD, curcumin can suppress Aβ accumulation, upregulate the expression of Akt, downregulate the expression of GSK3β, and inhibit the proinflammatory cytokine IL-1β. But in the late stage, curcumin has an insignificant inhibitory effect on GSK3β. In patients with AD, a low expression of Akt and a high expression of GSK3β were observed. Curcumin may have a similar effect on patients with AD by regulating these protein expressions and can be used to improve the pathological features of AD in the early stages of the disease.

Background: 20(R)-ginsenoside Rg3 (R-Rg3) is a kind of ginseng glycol tetracyclic triterpenoid saponin, which is a recognized traditional Chinese medicine monomer and has the action of inhibiting the proliferation of tumor cells. Objectives: The goal of our experiment was to investigate the protective function of R-Rg3 on chemotherapy-induced myelosuppression in mice. Materials and Methods: Cyclophosphamide (CTX) was injected into the intraperitoneal (i.p.) to establish the mice myelosuppression model. We measured the number of peripheral blood cells (PBCs), and the number of bone marrow nucleated cells (BMNCs) was counted. Hematopoietic progenitor cells (HPCs) were cultured in vitro, and the amount of cell colonies was recorded at different times. Then ELISA was used to detect the levels of hematopoietic-related cytokines and flow cytometry was employed to test cell cycle. The network pharmacology was used to predict the main pathways of action. Furthermore, the expression of p-JAK2 and p-STAT5 was detected via western blotting. Results: The experimental outcomes showed that R-Rg3 could improve the amount of PBCs in mice with myelosuppression, increase the quantity of karyota cells in bone marrow, and enhance the proliferation of HPCs and adjust the content of hematopoietic-related cytokines. Moreover, the JAK–STAT signaling pathway may be the key to the role of R-Rg3. Conclusion: All of these results implied that R-Rg3 might be a therapeutic agent for myelosuppression after chemotherapy.

Objectives: To investigate the effects of gavage and transdermal administration on the anti-tumor activity of QJ623 (extracts of Paris polyphylla rhizome and Tinosporae Radix) on a murine H22 solid tumor transplantation model. Materials and Methods: H22 cell suspensions were diluted to a density of 4 × 10⁵ cells/mL with sterile saline and then injected into 6-week-old mice subcutaneously (0.2 mL) into the right anterior axilla to create a murine model of liver cancer. Tumor tissues were collected after drug administration. The tumor growth-suppression rate was calculated, and the tumor tissues were stained with hematoxylin and eosin to detect necrosis. Terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling assay was performed to observe the apoptosis. The expression of p-ACK1, p-AKT, Bax, Bcl-2, and Caspase3 in the tumor tissues was detected by western blotting. Bio-chemical kits were used to detect the serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), creatinine (Cre), blood urea nitrogen (BUN), and uric acid (UA) for liver and kidney function analyses. Serum levels of IL-6, TNF-α, IFN-γ, and IL-10 were determined using enzyme-linked immuno-sorbent assay. Results: Gavage and transdermal administration of QJ623 at different concentrations reduced the tumor mass and volume to different degrees; promoted tumor cell apoptosis; decreased the expression levels of the p-AKT and Bcl-2 proteins; increased the expression of the apoptotic proteins Bax and Caspase3; increased the number of Th1 (IFN-γ) cells; decreased the number of Th2 (IL-4) and Treg cells; reduced the serum AST, ALT, BUN, UA, IL-6, TNF-α, and IFN-γ levels; and increased the IL-10 levels. Conclusion: Both gavage and transdermal administration of QJ623 showed anti-tumor effects by promoting tumor cell apoptosis, decreasing the level of inflammatory factors, increasing the number of Th1 (IFN-γ) immune cells, and decreasing the numbers of Th2 (IL-4) and Treg immune cells.

Background: Orostachys japonicus has traditionally been used for antifever, anti-inflammation, and anticancer. O. japonicus has not yet been studied for cosmetic effects. Objectives: In this study, O. japonicus extracts were investigated to verify the possibility of them as anti-aging cosmetic agents. Materials and Methods: O. japonicus were used to prepare experimental extracts using 90% methanol (MeOH) or ethanol (EtOH) as solvent. Safety and efficacy tests applied in this work are as follows: Folin-Denis' method for total phenolic contents, liquid chromatography for component analysis, 3-[4,5-dimethyl thiazol-2-yl]-5-(3-carboxymethoxyphenyl) -2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay for cytotoxicity, 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), and ferric reducing antioxidant power (FRAP) assay for antioxidant activity, biochemical inhibition assays of elastase activity, collagenase activity, matrix metalloproteinase-1 (MMP-1) and MMP-3 mRNA expression), and MMP-1 protein production for anti-wrinkle activity, and tyrosinase activity inhibition assay for whitening activity. Results: This research revealed that O. japonicus extracts contained various flavonoids, showed no adverse cytotoxicity up to 300 or 500 μg/mL in HS68 and B16F10, exhibited significant antioxidant activity, exerted remarkable anti-wrinkle activity, and represented useful whitening activity. Conclusion: This study suggested that O. japonicus extracts could be utilized to develop anti-aging cosmetic agents exerting useful whitening activity as well as excellent antioxidant and anti-wrinkle activities.

Objectives: The present study evaluated Ruta graveolens Linn. extract (RGLE) for its possible role in the treatment of vascular dementia (VD) in the rat model and inhibition of autophagy in hippocampus tissues. Materials and Methods: The spatial working memory of the rats was assessed using the established Tmaze tests. A video camera was used for recording and water maze software (HVS Image 2020; HVS Image Software Ltd.) was used for analysis of the digital images to measure escape latency and swimming distances for each rat. Results: RGLE treatment of the VD rats significantly (p < 0.05) alleviated the impairment in spontaneously altered behaviors and significantly (p < 0.05) reduced escape latency. The VD-mediated decrease in distance travelled, swim time, and count of platform crossings was significantly (p < 0.02) alleviated by RGLE treatment of the rats. In RGLE-treated rats, the VD-mediated increase in Beclin-1 and Microtubule-associated protein light chain 3II (LC3II) expression in the hippocampus tissues was significantly (p < 0.05) alleviated. RGLE treatment prevented the suppression of the mammalian target of rapamycin (mTOR) phosphorylation in VD rat hippocampus tissues. The rapamycin-mediated suppression of p-mTOR and elevation of Beclin 1 and LC3II expression in the rat hippocampus could not be alleviated by RGLE treatment. Conclusion: In summary, RGLE effectively prevents VD-mediated cognitive impairment and neuronal damage in the rats. Moreover, autophagy was inhibited and mTOR pathway was activated in rats with VD by RGLE treatment. Therefore, RGLE may be studied further as a therapeutic agent for treatment of dementia.

Background: Agarwood is a resinous heartwood produced in certain Aquilaria species that is often used as a spice and Chinese medicine materials. Objectives: This study aimed to comprehensively evaluate the agarwood quality of different species under the same inducer as well as different inducers of the same species. Materials and Methods: The GC-MS data of agarwood were retrieved by AMDIS to obtain 47 secondary metabolites. Combined with multivariate statistical analysis, 12 secondary metabolites were identified as potentially representing the differences between A. malaccensis and A. sinensis. The gray correlation degree and TOPSIS method were used to comprehensively grade 18 characters, including the AEC, the content of agarotetrol, the apparent abundance of GC-MS fingerprint, and the 15 secondary metabolites representing the 58 batches of samples. The OD values representing the overall desirability of r_i (gray correlation degree) and c_i (TOPSIS) were ranked, with a higher ranking reflecting, better agarwood quality. Results: The ranking results demonstrated that the agarwood samples of the top 33 in OD value were all induced by FAM71, whereas the agarwood samples of the top 23 were all from A. malaccensis. The agarwood of A. sinensis induced by FA had the lowest OD value. Conclusion: The study demonstrated that the quality of agarwood from A. malaccensis was better than that of A. sinensis using the same inducer of FAM71. In the same species of A. sinensis, the quality of agarwood produced by FAM71 was better than that induced by formic acid alone or NA8 alone. This study provided a theoretical basis for the selection of high-quality agarwood inducer and tree species, as well as a reference basis for the efficient production of agarwood in the actual production process.

Background: Cholangiocarcinoma (CCA), epithelial bile duct cancer, is deadly cancer with a very poor prognosis and standard anticancer drugs strategy remains poor and unfavorable outcomes. Objectives: In this study, we examined how Oroxylum indicum leaf extract in combination with gemcitabine (anticancer drug) affects the viability, apoptosis, and migration of CCA cells. Materials and Methods: Two CCA cells, KKU-100 and KKU-M452, were incubated with O. indicum or gemcitabine or in combination for 24-72 hr and these effects were explored by using the sulforhodamine B, colony formation, cell cycle arrest, ROS formation, mitochondrial function, migration, and Western blot analysis. Results: O. indicum expressively suppressed the growth of both CCA cells at 72 hr with IC₅₀ values were 9.88 ± 1.36 and 8.56 ± 1.02 μg/mL for KKU-100 and KKU-M452 cells, with the number of cells were decreased by dose-dependent manner. Further, the extract caused the reduction of colony formation and then suppressed the cells at G0/G1 phase for KKU-100 and S to G2/M phase for KKU-M452 cells. At 250 μg/mL, two CCA cells generated ROS formation along with decreasing mitochondrial function and then led to induce cancer cells apoptosis. Furthermore, O. indicum extract inhibited CCA cell migration via decreasing MMP-9 expression levels. The mechanism of its action was significant suppressed EGFR and caspase 3 levels. Combination group of O. indicum plus gemcitabine found that O. indicum potentiated gemcitabine to inhibit cell viability, induce apoptosis, increase ROS formation, decrease mitochondrial function, and suppress EGFR expression. Conclusion: O. indicum leaf extracts with potentiation of gemcitabine can reduce CCA cell growth, induce apoptosis, and cell migration through decreasing EGFR and caspase 3 expression. O. indicum could be useful for enhancing the activity of anticancer drugs displayed to prevent or treat CCA.

Aim/Background: We investigated the composition and structure of intracellular polysaccharides in cucumber seeds (CSs) and their effect on Sprague-Dawley (SD) rats and potential activity for fracture healing. Materials and Methods: Polysaccharides are its main water-soluble active ingredient. The SD rat tibia fracture model was established by surgical method, and the healing effect of cucumber seed polysaccharides (CSPs) on SD rat tibia fracture was preliminarily explored. Through histopathological observation, determination of rat serum indicators alkaline phosphatase (AKP), acid phosphatase (ACP), BGP, X-ray imaging observation, and micro-Computed Tomography (CT) imaging observation at different periods. Results and Conclusion: The results showed that CSP enhanced the activity of osteoblasts, increased the metabolic rate of osteoclasts, increased the secretion of AKP, BGP, and ACP, shortened the fracture healing time of SD rats and effectively promoted the healing of tibial fractures in SD rats.

Background: Thrombosis is a major complication and is responsible for cardiovascular diseases (CVDs). The majority of CVDs like stroke, coronary syndrome, and vascular ailments are tightly connected to venous/arterial blood clots. The present research work was planned to address the curative benefits of pinocembrin against thrombosis in the experimental animal model. Materials and methods: The male Wistar rats were employed in this research work and the thrombosis was provoked to the rats via electrical shock by standard method. Animals were treated with 25 and 50 mg/kg of pinocembrin orally. The 20 mg/kg of aspirin was used as a positive control. The level of thrombin-provoked platelet aggregation was performed using the standard method. The plasma coagulation parameters were examined using an automated blood coagulation analyzer. The status of tissue factor pathway inhibitor (TFPI), thromboxane-B2 (TX-B2), 6-keto-PGF1α, and TXB2/6-keto-PGF1α (T/K) was investigated using assay kits. The levels of urokinase-type plasminogen activator (u-PA), tissue-type plasminogen activator (t-PA), PAI-1, and t-PA/PAI-1 were quantified using assay kits. Results: The pinocembrin treatment appreciably prolonged the coagulation parameter time periods like activated partial thromboplastin time, thrombin time, and TP. Pinocembrin also elevated the TFPI, 6-Keto-PGF1α, u-PA, and t-PA status and reduced the platelet aggregation, TX-B2, T/K, PAI-1, and t-PA/PAI-1 levels in the experimental rats. Conclusion: The findings of this work suggested that the pinocembrin exhibited potent antithrombotic activity and it could be a potential candidate to treat the thrombosis in the future.

Objective: The aim of this study was to demonstrate the role of the total flavones of Abelmoschus manihot (TFA) in the reactive proliferation of astrocytes after cerebral ischemia in mice. Materials and Methods: The expression of glial fibrillary acidic protein (GFAP) and secretion of chondroitin sulfate proteoglycans (CSPGs) from astrocytes in brain tissues were used to evaluate the effect of TFA on the reactive proliferation of astrocytes after cerebral ischemia. Besides, we detected the activities of angiotensin-converting enzyme (ACE) and ACE2 and production of angiotensin (Ang)-II and Ang-(1–7) in the brain tissues. Furthermore, the role of Ang-(1–7) and TFA in GFAP expression and proliferation of primary cultured astrocytes under hypoxia induced by cobalt chloride (CoCl₂) was tested. Results: Cerebral ischemia induced a significant increase of GFAP expression and CSPGs secretion in mice brain tissues, which was inhibited by TFA treatment. In addition, TFA treatment inhibited the increment of ACE activity and Ang II production induced by ischemia in the brain tissues; likewise, TFA induced a significant upregulation of ACE2 activity and Ang-(1–7) production. Furthermore, TFA or Ang-(1–7) treatment markedly reduced reactive proliferation of astrocytes under hypoxia. Conclusion: TFA inhibits reactive proliferation of astrocytes in the brain tissues following cerebral ischemia by upregulating ACE2/Ang-(1–7).

Background: Psoralea Fructus (PF) is one of the most frequently applied tonic Chinese herbs with various bioactivities, while recently, the idiosyncratic drug-induced liver injury (IDILI) nature of PF was addressed, and liver injury cases were reported in the clinic. Our previous research indicated that many constituents exerted liver injury property, whereas the pharmacokinetic profile of these compounds between normal and immune stress states is still unclear. Objectives: This study aims to construct a validated method to simultaneously determine eight analytes (including psoralen, isopsoralen, psoralidin, corylin, psoralenoside, isopsoralenoside, bavachin, and neobavaisoflavone) in PF via UPLC-TQ/MS, and to compare their pharmacokinetic properties in normal and LPS-stimulated rats. Materials and Methods: The rats were randomly divided into four groups. The blood samples of different groups were harvested at different time points; the eight analytes were analyzed under UPLC-TQ/MS, and the constructed method was confirmed in terms of specificity, linearity, lower limit of quantitation (LLOQ), precision, accuracy, stability, recovery, and matrix effect. Results: The developed UPLC-TQ/MS method showed good linearity (r = 0.9972–0.9994) and specificity; the recovery and matrix effect were acceptable (ranging from 64.75 ± 1.59% to 104.31 ± 3.38%, and from 87.11 ± 1.91% to 115.45 ± 1.63%, respectively), the intra- and inter-day precision of eight analytes were under 13.42%, the intra- and inter-day accuracy was eligible (92.68%–108.85%), and the analytes were stable under storage conditions. All eight analytes in PFE were rapidly absorbed into the circulation, while the relevant pharmacokinetic parameters (including AUC, MRT, VRT, λ, V, t_1/2, C_max, T_max, and CL) of these analytes were quite different between the normal and model rats. Conclusion: A sensitive, accurate, and rapid method was successfully established and validated to determine the plasma characteristics of analytes in normal and LPS-primed rats. The pharmacokinetic profile indicated the body states might appreciably impact the pharmacokinetic profile of the bioactive constituents of PF, further inducing liver injury in specific patients.

Introduction: Erectile dysfunction (ED) is a common complication of diabetes mellitus (DM) that severely affects the patient's quality of life. Schisandrin A, maybe a novel therapeutic option for patients with diabetes mellitus-induced erectile dysfunction (DIED). Materials and Methods: After induction by streptozotocin administration, rats were divided into four groups: Normal control (NC), DIED, low dosage (5 mg/kg/d), middle dosage (10 mg/kg/d), and high dosage (20 mg/kg/d) of schisandrin A treated DIED group. All groups were treated with normal saline or the relevant drug for 8 weeks. Body weight, erectile rate, intracavernosal pressure (ICP), mean arterial pressure (MAP), and serum glucose concentration were measured. The nitrate reductase method and nitric oxide synthase activity assay detected nitric oxide (NO) and endothelial nitric oxide synthase (eNOS) levels. Pulldown assay detected GTP-RhoA activity. Western blotting detected alpha-smooth muscle actin (α-SMA), transforming growth factor-β1 (TGF-β1), Collagen III, Collagen IV, RhoA, ROCK1, ROCK2, phospho-myosin phosphatase target subunit 1 (p-MYPT-1), MYPT-1, phospho-eNOS (p-eNOS), and eNOS expressions. Results: Compared with the NC group, schisandrin A alleviated the ED rate of DIED rats, and increased ICP and ICP/MAP in DIED + SA group. Schisandrin A increased NO level and activated p-eNOS in penile tissues of DIED group. The expression of α-SMA increased, whereas, TGF-β1, Collagen III, Collagen IV decreased in DIED + SA group compared to DIED group. Schisandrin A inhibited the levels of GTP-RhoA, RhoA, ROCK1, ROCK2, and p-MYPT1 in penile tissues from rats of DIED. Schisandrin A in the high dose group (20 mg/kg/d) had a better effective effect on DIED. Conclusion: Schisandrin A ameliorated erectile dysfunction in rats with DIED by promoting eNOS production, inhibiting fibrosis, and inhibiting the RhoA/ROCK1 pathway.

Background: Tabebuia impetiginosa is an important medicinal plant rich in lapachol, α-lapachone, and β-lapachone known to possess several biological activities. Objective: In this study, we investigated the drug potential of lapachol, α-lapachone, and β-lapachone using molecular docking, molecular dynamic (MD), and drug-likeness properties. Materials and Methods: The computational study was performed using SwissADME software for the determination of the pharmacokinetic properties of the tested compounds. AutoDock Vina and Genetic Optimization for Ligand Docking (GOLD) were used for the docking analysis, and MD simulations were run using Schrodinger's Desmond Simulation. Results: The three compounds lapachol, α-lapachone, and β-lapachone binds to cysteine (Cys)-histidine (His) catalytic dyad (Cys145 and His41) along with the other residues with, respectively, the following docking score 48.69, 47.06, and 47.79. Against viral entry receptor, human angiotensin-converting enzyme 2 (hACE-2), α-lapachone exhibited the highest GOLD Fitness score complex (54.82) followed by lapachol (42.53) and β-lapachone and hACE-2 (38.74) generating several active sites in the target proteins. A 100 ns MDs simulation study revealed the stable conformation of bioactive compounds within the cavity of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) of hACE-2 protein and main protease (Mpro). From the dynamic study, it was observed that lapachol was tightly bound with catalytic dyad residue Cys145 of Mpro with more than 40% time of simulation, also post-simulation MM-GBSA binding free energy (ΔG Bind) revealed the highest energy score (−51.18 ± 5.14 kcal/mol) among the evaluated complex. Moreover, the absorption, distribution, metabolism, and excretion (ADME) properties demonstrated that the investigated compounds passed the pharmacokinetic and drug-likeness criteria without undesirable effects. Conclusion: The computational study highlighted that these compounds could be highly recommended and developed as part of an effective drug against the SARS-CoV-2 virus.

Background: Angelica gigas Nakai (AGN) is well-known for anti-inflammatory and anti-oxidative effects; however, the mechanism underlying the protective effects of AGN on monosodium iodoacetate (MIA)-induced osteoarthritis (OA) remains unknown. Objectives: Our objectives of this study were to evaluate the anti-inflammatory effect of AGN on MIA-induced OA and investigate the potential mechanism which underlies such effect. Materials and Methods: The dried herb AGN was extracted with distilled water. After the experiment was conducted for 4 weeks using 7-week-old male Sprague–Dawley rat. AGN (200 mg/kg body weight/day) was orally administered for 14 days from day 7 after intra-articular injection of MIA (50 μL with 80 mg/mL), and the effects were compared with those of MIA-treated control group. We examined the weight-bearing ability of hind paws, biomarkers related to oxidative stress in cartilaginous tissue. In addition, various factors associated with inflammatory and cartilage degeneration in cartilaginous tissue detected by the Western blot analysis. Results: Our results from an in vivo model of OA indicate that AGN inhibits inflammatory factors such as inducible nitric oxide synthase, cyclooxygenase-2, tumor necrosis factor-α, interleukin-6 (IL-6), and IL-1β via nuclear factor kappa B signaling pathway. AGN suppresses the expression of matrix metalloproteinase, proteolytic enzyme, and also elevates tissue inhibitor of metalloproteinase which inhibited degradation of the extracellular matrix. Moreover, antioxidant defense was enhanced through the nuclear factor erythroid 2-related factor 2/heme oxygenase-1 pathway. Conclusion: Taken together, these results demonstrate the effectiveness of AGN at the inflammation associated with joint disorders. Our results also suggest that AGN could serve as a potential candidate for alternative therapeutic treatment of OA.