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   2020| Jan-Mar  | Volume 16 | Issue 67  
    Online since February 11, 2020

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Evaluation of lipopolysaccharide-induced acute lung injury attenuation in mice by Glycyrrhiza glabra

Jan-Mar 2020, 16(67):92-98
Objective: The medicinal importance of Glycyrrhiza glabra is well reported in Asian folk core medicine. In the current study, we have investigated the treatment effects of ethanolic extract of G. glabra in acute lung injury (ALI) mice models. Materials and Methods: ALI murine models were established by intratracheal instillation of bacterial lipopolysaccharide (LPS). Bronchoalveolar lavage fluid (BALF) was prepared for measuring cell migration and cell count, protein content, and superoxidase dismutase (SOD) activity. Lung tissues were collected for evaluating the wet-to-dry weight ratio, messenger ribonucleic acid (mRNA) expression of pro-inflammatory cytokine, and histological change. Results: It was observed that the treatment of ALI mice significantly decreased the total cell count and exudation of protein into BALF. Further, SOD activity in BALF of LPS-induced ALI mice was greatly improved when treated with G. glabra extract at 200 or 400 mg/kg of body weight, and the SOD activity increased in a dose-response manner. Evaluation of lung wet-to-dry weight ratio, pro-inflammatory mRNA expression levels, and histologic examination of lung tissues indicates that plant extract has significantly attenuated the tissue injury. Conclusions: The results showed that G. glabra had a protective effect on LPS-induced ALI in mice which might be contributed due to diverse phytoconstituents of the plant extract. To fulfill the unmet needs of treating ALI, the active principles in G. glabra could be promising lead-alternative therapy to develop high-potent anti-inflammation agents for ALI.
  1,242 196 -
Effects of baicalin on pharmacokinetics of florfenicol and mRNA expression of CYP1A2, CYP2C11, CYP3A1, UGT1A1, MDR1, and ABCC2 in rats
Si-Cong Li, Xu-Ting Li, Bin Wang, Rui Yang, Min Zhang, Jin-Liang Li, Wei Huang, Liang Cao, Song-Yang Xiao
Jan-Mar 2020, 16(67):1-6
Background: The traditional Chinese medicine Scutellariae radix is often used in combination with antibiotics, such as florfenicol, in Chinese veterinary medicine. Baicalin (5, 6, 7-trihydroxyflavone-7-β-D-glucuronide) is the main active constituent of S. radix. The effects of baicalin on the pharmacokinetics of florfenicol are not known. Thus, we have studied the effects of baicalin on florfenicol pharmacokinetics and the mRNA expression of drug-metabolizing enzymes/efflux transporters in rats. Materials and Methods: Sprague-Dawley rats were given baicalin (50 mg/kg BW or 100 mg/kg BW) or 0.9% sodium chloride solution by intragastric administration for 7 days. They were then fed florfenicol by intragastric administration (25 mg/kg BW) on the 8th day. Blood samples were collected at various times, and the plasma concentrations of florfenicol were estimated with ultra-high performance liquid chromatography. The mRNA expression levels of cytochrome P450 (CYP) CYP1A2, CYP2C11, CYP3A1, UDP-glucuronosyltransferase 1A (UGT1A), multidrug resistance 1, and ATP-binding cassette C2 (ABCC2) in the liver and jejunum were analyzed with a real-time polymerase chain reaction. Results: The area under the concentration-time curve from zero to infinity, mean residence time from zero to infinity, elimination half-life, and peak concentration of florfenicol were significantly increased and the apparent volume of distribution fraction of the dose absorbed and plasma clearance fraction of the dose absorbed were significantly decreased by baicalin; the mRNA expression levels of CYP1A2, CYP2C11, CYP3A1, UGT1A1, and ABCC2 were down-regulated by baicalin. Conclusion: Baicalin affected the pharmacokinetics of florfenicol in rats, increased the plasma concentration and residence time of florfenicol, probably by decreasing the mRNA expression levels of CYP1A2, CYP2C11, CYP3A1, UGT1A1, and ABCC2 in the liver and jejunum.
  917 444 -
Antioxidant effect of Terminalia arjuna extract against acetaminophen-induced hepatotoxicity via the regulation of cytochrome P450 2E1, phosphatidylinositol-3-kinase/protein kinase B
Senthilganesh P Kannappan, Gunapriya Raghunath, Senthilkumar Sivanesan, Rajagopalan Vijayaraghavan, Madhankumar Swaminathan
Jan-Mar 2020, 16(67):13-20
Aim: The present study explored the therapeutic in detail antioxidants and effect of aqueous Terminalia arjuna (TA) bark extract against acetaminophen (APAP) induced hepatotoxicity through studies on serum marker enzymes, phosphatidylinositol3kinase/protein kinase B (PI3K/AKT) pathway, CYP2E1 evaluations. Biochemical, antioxidant, cytochrome P450 2E1 (CYP2E1) enzyme, and PI3K/AKT cell signal enzymes were observed with the appropriate methods of study. Materials and Methods: The animals were divided into five groups (each having six animals): control group, Acetaminophen (APAP) toxic group, N-acetylcysteine (NAC) group, TA 250 mg/kg group, and TA 500 mg/kg group. APAP toxic dose of 750 mg/kg body weight was administered along with 0.5% of hydroxypropyl cellulose (vehicle) 24 h before sacrificing the animal. Results: The biochemical, antioxidant, Histopathological, CYP2E1 enzyme, PI3K, AKT protein expression analysis were shown increased antioxidant level, increased PI3K/AKT level, decreased liver function marker level and decreased CYP2E1 level in TA500 mg group compared with APAP toxic group (P < 0.01). The findings suggest that TA (500mg/kg) drug reduced Acetaminophen toxicity via antioxidant and molecular mechanisms. Conclusion: The present study concluded that TA 500 mg/kg high dose is more effective to restore the liver tissue through APAPinduced hepatotoxicity in Wistar albino rats.
  1,016 308 -
Phytochemical profile of the aerial parts of Rehmannia glutinosa liboschitz var. purpurea Makino
Huu Tung Nguyen, Thi Ngan Dan, Takuhiro Uto, Tomoe Ohta, Hitoshi Watanabe, Yukihiro Shoyama
Jan-Mar 2020, 16(67):128-131
Background: The roots of Rehmannia glutinosa (RG) or Rehmanniae Radix are a well-known medicinal material in the Oriental medicine, and its phytochemical profile has been extensively studied with more than 100 individual compounds from Rehmannia species. In contrast, bioactive components of the aerial part of the title plant are largely unknown as only several compounds reported up to date. Objective: The objective was to study on chemical constituents of the aerial parts of the title plant and evaluate the aerial parts as a supplementary source for Rehmanniae radix. Materials and Methods: Solvent extraction, partition, and column chromatography was used to separate individual compounds; spectroscopic data including nuclear magnetic resonance and mass spectrometry were analyzed to determine the chemical structure of the isolates. Results: Eight compounds including five ursane-type triterpenoids for the first time from RG (ursolic acid [1], pomolic acid [2], 2β-hydroxypomolic acid [3], asiatic acid [4] and 7β,24-dihydroxy ursolic acid [5]) and three main glycosides (ajugol [6], aucubin [8], and acteoside [7]) were characterized from the aerial parts of the title plant. Their structures were identified on the basis of spectroscopic data and comparison with those reported in the literature. Conclusion: The current study reveals various ursane triterpenes in the organic portion beside the main hydrophilic glycosides in the RG aerial parts. The occurrence of various ursane triterpenes contributed in part to phytochemical database and evidence of the biological activity associated with potential in use as a medicinal material of the RG leaves.
  1,057 172 -
Protective effect of ferulic acid on human umbilical vein endothelial cell model of cold stress

Jan-Mar 2020, 16(67):7-12
Context: Ferulic acid (FA) is an active principle derived from the traditional Chinese medicine Angelica sinensis, which has been used for the treatment of cardiovascular and cerebrovascular diseases in China for many years. However, a thorough understanding of effects on vascular function by FA has not been investigated. Aims: The aim of the present study was to investigate the potential mechanism of FA by suppressing Transient receptor potential cation channel subfamily M member 8 (TRPM8) channels and regulating endothelial nitric oxide (NO) pathway to ameliorate cold explore injury in human umbilical vascular endothelial cells (HUVECs). Subjects and Methods: The effects of cold exposure and FA on cell viability were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and lactate dehydrogenase (LDH) assay. Quantitative polymerase chain reaction and Western blot were utilized to detect TRPM8, hypoxia-inducible factor-alpha (HIF-1α), endothelin-1 (ET-1), inducible NO synthase (iNOS), endothelial NO synthase (eNOS) messenger RNA, and protein expression in HUVECs. Enzyme-linked immunosorbent assay method was used to detect the concentration of ET-1 in culture supernatants of HUVECs. Results: Cold exposure at 18°C had no significant effect on cell morphology but increased secretion of LDH and ET-1 and the expression of TRPM8, HIF-1α, iNOS, and ET-1. Treatment with FA decreased all of these changes. The levels of NO and eNOS decreased in cold stress model, while FA treatment attenuated the cold-induced decrease of NO and eNOS. Conclusion: Cold stress can cause an increase in vasoconstrictors such as TRPM8 and ET-1 and reducing cell viability, but FA can prevent cold stress-related cardiovascular disease by regulating the expression of these substances in cells.
  908 237 -
Constituents of flavonoids from Tridax procumbens L. and antioxidant activity

Jan-Mar 2020, 16(67):201-205
Background: To study the chemical constituents and antioxidant activities of flavonoids in Tridax procumbens L. Materials and Methods: Diaion HP-20, MCI-Gel CHP-20, Sephadex LH-20, and silica gel column chromatography were used to isolate and purify the chemical components of T. procumbens L., and the structures of the compounds were identified by physical and chemical properties and spectral techniques (1H-nuclear magnetic resonance [NMR] and 13C-NMR). The antioxidant activities of five flavonoids were evaluated by 2,2-diphenyl-1-picryl-hydrazyl method. Results: Six compounds were isolated and identified as luteolin-7-O-β-D-glucoside (1), 5,7,3'-trimethyl-4'-methoxyl-3-O-β-D-flavonoid glucoside (2), 8,3'-dihydroxyl-3,7,4'-trimethoxy-6-O-β-D-flavonoid glucoside (3), 4,2',4'-trihydroxy-6'-methoxyl-3'-isopentenyl chalcone (4), quercetin-7-O-β-D-glucopyranosyl-(2→1)-α-L-rhamnose (5), and nodakenin (6). Conclusions: All compounds except compound 3 were isolated from the plant for the first time. It was found that compounds 1–5 had significant antioxidant and free radical scavenging activities.
  915 204 -
Extraction, purification, content analysis and hypoglycemic effect of mulberry marc anthocyanin

Jan-Mar 2020, 16(67):68-75
Background: Mulberry marc (MM) which as a wasted residue contains many available resources and one of the valuable nutrients is anthocyanin. In this study, the extraction, purification, content analysis, and hypoglycemic effect of anthocyanin from MM were studied. Materials and Methods: The total anthocyanin and cyanidin-3-O-glucoside of the purified anthocyanin of mulberry marc (MMA) were determined using pH differential method and high-performance liquid chromatography. The hypoglycemic effects were studied by streptozotocin and high-fat diet-induced type 2 diabetic rats. The rats were given MMA for 4 weeks and at the end of experiment; the blood was collected for determination serum indexes; the tissue of liver, kidney, pancreas, and spleen was removed for weighting and observing pathological changes. Results: The content of total anthocyanin and cyanidin-3-O-glucoside of MMA was 39.72% and 18.63%, respectively. The fasting blood glucose of rats in the MMA treatment groups was decreased. The lipid metabolites (triglycerides and total cholesterol) and malondialdehyde were decreased and superoxide dismutase was increased of type 2 diabetes rats after MMA treated; the pathological changes were alleviated in various degrees. Conclusion: The content of anthocyanin in mulberry marc is higher than mulberry juice, and the MMA has a good hypoglycemic effect.
  911 145 -
Skin-protective and anti-inflammatory effects of Hibiscus syriacus L. (Mugunghwa): A comparative study of five parts of the plant
Jung-Eun Yang, Sul Woong Park, Hien T T. Ngo, Seul A Seo, Eun Byeol Go, Jeong-Seung Hwang, Eunson Hwang, Tae-Hoo Yi
Jan-Mar 2020, 16(67):183-191
Aim: The skin barrier is vulnerable to internal and external elements. However, certain plant extracts have promise for protecting the skin. Hibiscus syriacus L. is a Korean national plant (mugunghwa) that has been traditionally used to treat some skin diseases; however, its pharmacological activities are poorly understood. The present study has evaluated the skin-protective effect of mugunghwa flowers (MF), cortexes (MC), roots (MR), leaves (ML), and seeds (MS) in external factor-stimulated human epidermal keratinocytes (HaCaTs), normal human dermal fibroblasts (NHDFs), and a widely used murine macrophage cell line (RAW 264.7 cells). Materials and Methods and Results: The active components were identified as palmitic acid and linoleic acid by high-performance liquid chromatography analysis. The parts of mugunghwa had low 2,2-diphenyl-1-picrylhydrazyl scavenging activities but moderate 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt scavenging capacity. All parts' extracts at 100 μg/mL decreased nitric oxide production in inflammation-induced RAW 264.7 cells and mRNA expression of the cytokines interleukin-6/1β and the enzymes inducible nitric oxide synthase/cyclooxygenase-2. On HaCaTs, MF, MC, and MR (10–50 μg/mL) significantly increased hyaluronan production over that of controls. Moreover, MF activated NAD (P) H:quinone oxidoreductase 1 and heme oxygenase-1 and inhibited intracellular reactive oxygen species production. Treatment with MF, MC, MR, ML, and MS (10 μg/mL) reduced the scratching wound site in NHDFs to be considered by vascular endothelial growth factor signaling activation. Conclusion: Taken together, among all the five parts of the mugunghwa plant, MF effectively inhibited lipopolysaccharide, ultraviolet B irradiation, and scratching wound-induced inflammation in skin cells.
  842 193 -
Enhancing glucose uptake by Astraeus odoratus and Astraeus asiaticus extracts in L6 myotubes
Papawinee Phadannok, Alisa Naladta, Kusumarn Noipha, Natsajee Nualkaew
Jan-Mar 2020, 16(67):34-42
Background: Mushrooms, including Astraeus spp., are known for hypoglycemic properties. Astraeus odoratus is a delicious edible mushroom, while Astraeus asiaticus is less popular due to its unpleasant texture. Both mushrooms have not been reported for the glucose uptake activities yet. Objectives: The aim of this study is to describe the enhancing of glucose uptake and related mechanisms in vitro of the extracts from two Astraeus spp. Materials and Methods: The extracts of A. odoratus and A. asiaticus (AO and AA, respectively) were assayed for the stimulation of glucose uptake in L6 myotubes. The mechanism of actions was proved by using specific inhibitors and determined for the expression of glucose transporters type 1 and 4 (GLUT1 and GLUT4) by quantitative real-time polymerase chain reaction and Western blotting. Results: The extracts of both mushrooms enhanced glucose uptake in the muscle cells L6 myotubes at the level of the function of GLUT1 and GLUT4, which involved the partial stimulation of their intrinsic activities through p38 mitogen-activated protein kinase and increased GLUT1 and GLUT4 protein levels. AO increased both GLUT1 and GLUT4 protein, while AA increased mainly GLUT4 protein and stimulated GLUT4 translocation through phosphatidylinositol 3-kinase. Conclusion: The results supported the hypoglycemic activity of A. odoratus and A. asiaticus and suggested their potential use for hypoglycemic purposes.
  879 133 -
Anti-inflammatory and antinociceptive activities of Salvia keerlii
Roberto Serrano-Vega, Cuauhtemoc Pérez-González, Ángel Josabad Alonso-Castro, Juan RamRamón Zapata-Morales, Salud Pérez-Gutiérrez
Jan-Mar 2020, 16(67):27-33
Background: Inflammation is a response to an attack on the body and is treated with several drugs that cause severe side effects. An alternative treatment source is plants, such as Salvia keerlii (SAKE) Benth. Objective: The objective is to study the determination of the anti-inflammatory and antinociceptive properties and chemical composition of the chloroform extract of SAKE. Materials and Methods: The plant was defatted with hexane, and then, chloroform and methanol extracts were obtained by heating, and the solvent was evaporated. The chemical composition of the extract was analyzed using gas chromatography–mass spectrometer. Total phenolic was determined using Folin–Ciocalteu reagent, and total flavonoid was determined by AlCl3assay. The acute and chronic anti-inflammatory effects of SAKE were evaluated on ear edema induced with 12-O-tetradecanoylphorbol-13-acetate, and the antinociceptive activity was determined. The effects of SAKE on protein denaturation and membrane stabilization were determined. The levels of nitric oxide, tumor necrosis factor-alpha (TNF-α), interleukin (IL)-1β, IL-6, and IL-10 were measured in J774A.1 macrophages stimulated with lipopolysaccharide. Results: SAKE at 2 mg/ear inhibited ear edema by 84.96% in the acute assay, whereas 100 and 200 mg/kg (p. o.) SAKE decreased inflammation in the chronic assay. In macrophages, SAKE decreased the levels of TNF-α (1.7-fold), IL-1β (1.7-fold), and IL-6 (1.9-fold) but increased the levels of IL-10 (1.9-fold). SAKE inhibited protein denaturation (IC50 =8.68 μg/mL). The inhibition of writhing obtained with SAKE at doses of 10, 50, 100, and 200 mg/kg was 28.73%, 46.48%, 56.1%, and 63.05%, respectively. Conclusions: SAKE has in vitro and in vivo anti-inflammatory and antinociceptive effects.
  758 219 -
Red seaweed (Eucheuma cottonii) extract promotes human keratinocyte migration via p38 mitogen-activated protein kinase phosphorylation
In Wook Kim, So Yeon Kim, Marianti Manggau, Harfina Finanda Anwar, Nyoun Soo Kwon, Kwang Jin Baek, Hye-Young Yun, Dong-Seok Kim
Jan-Mar 2020, 16(67):192-196
Background: Eucheuma cottonii (EC) is a rapidly growing red seaweed in Southeast Asia. Objectives: This study was performed to investigate the effects of EC extract on human keratinocyte migration. Materials and Methods: Cell migration was assessed using the scratch wound assay. Western blot analysis was performed to investigate the EC extract-induced signaling pathways. Results: EC extract promoted HaCaT keratinocyte migration in a concentration-dependent manner. To investigate the mechanism of EC extract-induced migration, we examined the migration-related signaling pathways. Western blot analysis showed that the EC extract showed no changes in extracellular signal-activated kinase (ERK) but showed slight Akt activation. In contrast, the EC extract strongly phosphorylated p38 mitogen-activated protein kinase (MAPK) and the subsequent downstream molecule, cAMP response element-binding protein (CREB). To examine the involvement of p38 MAPK pathway in EC extract-induced migration, SB203580, a specific inhibitor of p38 MAPK, was used. The results showed that EC extract-induced migration was abrogated by SB203580 pretreatment. In addition, SB203580 also blocked p38 MAPK and CREB activation. Conclusion: Taken together, these data suggest that EC extract promotes migration in human keratinocytes via p38 MAPK and CREB phosphorylation.
  837 135 -
The effect of Phyllanthus debilis methanolic extract on DNA methylation of TAC1 gene in colorectal cancer cell line

Jan-Mar 2020, 16(67):57-60
Aim/Background: In colorectal cancer, TAC1 was shown to be highly methylated in early carcinogenesis. Using HT29 cell line as a model of colorectal cancer, we postulated that Phyllanthus debilis methanolic extract could regulate DNA methylation in TAC1 gene promoter region and thus could alter the progression of colorectal cancer. Methodology: Cell culture study was done using HT29 cells, which were treated with 0.117 mg/ml P. debilis methanolic extract or 0.5 μM 5-Aza-2-Deoxycytidine (5-Aza) for 72 h. Cells were harvested at 72 h and were extracted for DNA. The DNA was bisulfite modified before been PCR and later was pyrosequenced. Results: The treatment with P. debilis significantly decreased the DNA methylation of TAC1 gene at site 1 (93.1% ± 2.4% vs. 100% ± 0.1%, P < 0.05), site 2 (86.5% ± 1.3% vs. 91.7% ± 0.2%, P < 0.05), and site 3 (96.7% ± 0.9% vs. 100% ± 0%, P < 0.05); but, no significant changes of DNA methylation were seen at site 4 (96.5% ± 1.9% vs. 93.6% ±0.6%, P > 0.05). The average of all Cytosine nucleotide followed by Guanine nucleotide (CpG) sites methylation was reduced, but not statistically significant when compared to the untreated cells (mean methylation 93.2% ± 2.4% vs. 96.3% ± 2.2%, P > 0.05). For cells treated with 5-Aza, DNA methylation was significantly decreased only at site 2 (88.6% ±0.8% vs. 91.7% ±0.2%, P < 0.05), but there was no significant methylation changes at site 1 (94.8% ± 2.4% vs. 100.0% ±0.06%, P > 0.05), site 3 (98.9% ± 1.1% vs. 100% ± 0%, P > 0.05), site 4 (90.6% ±1.6% vs. 93.6% ±0.6%, P > 0.05), and the average of all CpG sites (mean methylation 93.2% ± 2.3% vs. 96.3% ± 2.2%, P > 0.05). Conclusion: Treatment of methanolic extract of P. debilis reduces the methylation of promoter region of TAC1 gene, with better effect than low dose 5-Aza at 72 h of treatment. The anticancer effect of P. debilis may be partly been regulated through DNA methylation.
  847 117 -
Cytotoxicity, phagocytic activity, and leishmanicidal potential of extract standardized in geranylgeraniol obtained from the fruit of Pterodon emarginatus vogel
Suzana Ferreira Alves, Clayson Moura Gomes, Matheus Gabriel de Oliveira, Wanessa Machado de Andrade, Larissa Cleres Moreira, Leonardo Luiz Borges, Cláudio Carlos da Silva, Graciella Ottoni de Souza, Vinicius Barreto da Silva, Marize Campos Valadares, Maria Teresa Freitas Bara
Jan-Mar 2020, 16(67):140-147
Aim: The propouse of this study was to available the cytotoxicity, phagocytic activity, and leishmanicidal potential of extract standardized from the fruits of Pterodon emarginatus Vogel. Background: P. emarginatus is a Brazilian medicinal plant with several constituents shown to have anti-inflammatory, antinociceptive, antimicrobial, and antiproliferative activities. Materials and Methods: The standardized extract in geranylgeraniol obtained from the fruit of P. emarginatus was investigated for clastogenic and cytotoxic effects, as well as phagocytic activity against Leishmania guyanensis. Results: A 24-h exposure of Allium cepa roots to standardized geranylgeraniol extract (9.77 mg/mL) promoted clastogenic effects, with a mitotic index (MI) of 10.33 ± 0.61. Extending the exposure time to 48 or 72 h significantly reduced the MI (P < 0.05). The IC50for geranylgeraniol in 3T3 and RAW 264.7 cells was found to be 15.5 mg/mL and 12.2 mg/mL, respectively. The standardized extract in geranylgeraniol exhibited high phagocytic and microbicidal activities in Leishmania, promoting a >60% increase in macrophage infection, with greater internalization and destruction of the parasite. In silico study results suggest that geranylgeraniol acts in Leishmania by interfering with the biosynthesis of sterols through steric hindrance of the active site of the enzyme CYP51. Conclusion: The standardized extract in geranylgeraniol may be useful as an antiproliferative agent, macrophage immunomodulator, and as a cytotoxic treatment against infectious agents.
  751 137 -
Role of Saccharum Granorum as a “Principal Drug” in a traditional Chinese medicine formula against chronic atrophic gastritis rats
Guohong Wang, XueMei Qin, Yuetao Liu
Jan-Mar 2020, 16(67):43-50
Background: Saccharum Granorum (YiTang) is a nutrient-rich food that has been used as traditional Chinese medicine (TCM). However, the mechanism responsible for its beneficial efficacy is poorly understood, especially for its roles involved in the TCM formulas. Materials and Methods: In the present work, a classical TCM formulas, Huangqi Jianzhong Tang (HQJZ), was selected to interpret the efficacy and “principal drug” role of YiTang against chronic atrophic gastritis (CAG) rats using a serum metabonomic approach. Results: Twelve candidate metabolites were identified to characterize the difference between CAG and normal rats, which were involved into four disturbed metabolic pathways, including glucose metabolism, amino acid metabolism, fatty acid metabolism, and choline metabolism. The further partial least square regression analysis revealed that five metabolites, including choline, acetate, alanine, a-glucose, and β-glucose, were considered as potential biomarkers related to CAG. YiTang could exert the synergistic effect with HQJZ without YiTang, where the whole formula obtained the best beneficial treatment against metabolic disturbance induced by CAG. The metabolic improvement of YiTang and HQJZ might exert the effects on the dysfunction of energy metabolism, gastric emptying, and the changes of gut microbiome, which were the important pathomechanisms induced by CAG. Conclusion: These findings suggested that YiTang played an indispensable role in HQJZ formula against CAG, which may provide an approach to understand the fundamental idea behind formula construction.
  738 128 -
Fucoxanthin attenuates behavior deficits and neuroinflammatory response in 1-methyl-4-phenyl-1,2,3,6 -tetrahydropyridine-induced Parkinson's disease in mice
Guoqing Sun, Tao Xin, Rui Zhang, Chengxiao Liu, Qi Pang
Jan-Mar 2020, 16(67):51-56
Background: Parkinson's disease (PD) is one of the foremost neurological disorders which is differentiated next to the progressive dopamine (DA) loss, especially in the area of substantia nigra pars compacta (SNpc). Aberrant neuroinflammation, as well as excessive reactive oxygen species (ROS) generation, have exposed to stimulate neuronal defeat in the gradual developing PD. The current study investigated whether fucoxanthin could attenuate the pathophysiology seen in an 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-mediated PD model. Materials and Methods: C57BL/6 mice received 30 mg/kg of MPTP (i.p.) every day for 5 consecutive days to establish PD, subsequently everyday treatment with fucoxanthin intended for 7 days. Intraperitoneal injection of MPTP, resulted in impaired motor functions, dopaminergic neuronal loss, decreased DA, Tyrosine hydroxylase (TH) levels, and microglial activation-mediated neuroinflammation. Results: Fucoxanthin administration ameliorated α-synuclein abnormal accumulation and motor impairment in MPTP-induced chronic PD mouse model. Our observed findings revealed that fucoxanthin administration reverses the MPTP-mediated decline of DA neuron, TH along with microglial activation predominantly in the SNpc region. The Western blot analysis exposed that fucoxanthin suppressed the expression patterns of proinflammatory cytokines following MPTP administration. Conclusion: In conclusion, Fucoxanthin exerts neuroprotective potential in opposition to MPTP-mediated PD mice through repressing α-synuclein expression, oxidative stress and gliosis, recommended that fucoxanthin might perform as a beneficial remedy toward PD amelioration.
  691 147 -
A rapid and efficient approach based on ultra-high liquid chromatography coupled with mass spectrometry for identification in vitro and in vivo constituents from shizao decoction
Yu-Mei Wang, Qi Liu, Wen-Hao Fu, Ai-Hua Zhang
Jan-Mar 2020, 16(67):148-155
Introduction: Shizao decoction (SZD) is a classic Chinese prescription which used in multiple kinds of ascites and edema caused by liver cirrhosis, chronic nephritis, etc. However, considering edema and new application of cancer ascites SZD is worth further investigating. Materials and Methods: In this paper, a powerful ultra-high liquid chromatography coupled with electrospray ionization quadrupole time-of-flight mass spectrometry based on the PeakView tool combined with multivariate data processing approach was employed to clarify the chemical constituents and the ingredients absorbed into blood after oral administration of SZD. Results: As a result, a total of 91 compounds (35 ions in positive mode and 56 ions in negative mode) of SZD in vitro were successfully identified. Besides, 25 constituents absorbed into blood were tentatively characterized. Among the characterized ingredients, flavonoids, prenol lipids, coumarins, and derivatives and benzopyrans were also detected, respectively. Of note, some constituents absorbed into blood played a critical role in cancer therapy. Conclusion: Established method was suitable for the rapid analysis and characterization of SZD constituents, and it provided meaningful chemical information for further pharmacology study of SZD.
  688 129 -
Influence of spectral light composition on flavones formation in callus culture of Scutellaria baicalensis georgi
Anna Yurievna Stepanova, Alexandra Ivanovna Solov'yova, Svetlana Andreevna Salamaikina
Jan-Mar 2020, 16(67):156-160
Background: Scutellaria baicalensis is one of the most popular medicinal plants, which roots extracts are widespread in medicine and cosmetology. The area of growth of the S. baicalensis is rapidly declining; therefore, the involvement of biotechnological approaches to obtain its biomass is relevant. Since the content of flavones in cultured in vitro cells is usually much lower than in intact plants, there is a need to strengthen the synthesis of target substances. Objectives: The objective of this study is to investigate the effect of blue, white, and red light for the growth and content of the four main flavones (baicalin, wogonoside, baicalein, and wogonin) in the callus tissue of the S. baicalensis. Materials and Methods: Calluses in the experiments were continuously illuminated for a month with blue (420–480 nm), red (600–650 nm), warm white (400–800 nm) using LEDs with illumination of 1 μmol/m2/s. High-performance liquid chromatography was used to determine the content of the main flavones. Results: The presence of the light of all studied parts of the spectrum contributed to the elongation of the stationary phase against the background of callus growth suppression, in addition, the content of flavones increased, mainly due to baicalin. The maximum number of flavones was formed in blue light (5%). Conclusion: The blue light is an important factor to the accumulation of the main flavones in the calluses of S. baicalensis. The results obtained can be used not only in fundamental research but also in practice.
  681 135 -
Vanillin an active constituent from Vanilla bean induces apoptosis and inhibits proliferation in human colorectal adenocarcinoma cell line
Yandong Xie, Jingyu Zhang, Cheng Wang, Qinglin Fan, Yiling Zhang
Jan-Mar 2020, 16(67):197-200
Background: Colorectal cancers global incidence is increasing due to the rapid acquisition of Western lifestyle habits such as high caloric nutrition and sedentary status. Natural bioactive products play critical roles in anticancer drug development. Aim: The aim of the study was to investigate the antiproliferative and proapoptotic effects of vanillin (VL) an active constituent from Vanilla bean in colon cancer (human colorectal adenocarcinoma cell line [HT-29]) cells. Methodology: We demonstrated the cell growth and apoptosis in HT-29 cells. The apoptotic measures were analyzed by analyzing mitochondrial membrane potential (Δψm) level; apoptotic bodies by acridine orange/ethidium bromide and Hoechst staining. Results: Our results indicated that VL induces apoptosis as evidenced by cell viability loss, resulting in the altered Δψm in HT-29 cells. Further, the prooxidant role of VL inhibit B-cell lymphoma 2 (Bcl2) expression with the simultaneous upregulation of BCL-2 associated X, Cytochrome c, Capspase-9 and 3 protein expressions in HT-29 cells. Conclusion: Thus, in this regard, we have concluded that VL induces apoptosis in HT-29 cells through inducing oxidative damage and modulates apoptotic marker expressions. VL may therefore be used as a therapeutic agent for colon cancer treatment.
  684 129 -
In vivo antimalarial potential of tinospora crispa miers in mice and identification of the bioactive compound

Jan-Mar 2020, 16(67):76-82
Background: Malaria is a serious disease that is causing huge toll on human health. The parasites causing this disease have developed resistance toward the current mainstream drugs. Tinospora crispa Miers is a well-known ethnopharmacological plant of Malaysia and has been traditionally used to treat malaria, fever, and other parasite-related illnesses. Objective: The present study investigated the potential of the crude methanol extract and antiparasitic fractions of T. crispa stem to exert antimalarial activity against Plasmodium berghei. Materials and Methods: The potent antiparasitic fractions of T. crispa, F4, and F5 were isolated in the previous study against Toxoplasma gondii. The same antiparasitic fractions along with crude methanol extract of different doses (10, 50, 100 mg/kg b. w.) were employed in the present study to determine the antimalarial activity against P. berghei ANKA strain. The survival curves of the treated mice were plotted by employing the log-rank (Mantel–Cox) test. The chemical composition of the most potent fraction F5 was determined spectrometrically using electrospray ionization-mass spectrometry (MS). Results: In a murine P. berghei model, fraction F5 displayed the highest parasitemia suppression effects compared to the crude methanol extract and other fraction. Subsequent chemical analysis by MS on fraction F5 has led to the tentative identification of 13-hydroperoxyoctadeca-9, 11-dienoic acid (13[S]-HPODE) compound. Conclusion: The crude methanol extract of T. crispa and its fraction F5 possess potent antimalarial activities, and the tentative discovery of the 13(S)-HPODE bioactive compound may serve as a precursor for developing a semisynthetic antiparasitic drug with enhanced efficacy and low toxicity.
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Physiological mechanism and developmental events in differentiating floral buds of Sophora tonkinensis gagnep

Jan-Mar 2020, 16(67):83-91
Background: Sophora tonkinensis Gagnep is an important medicinal plant in China. Previous research has focused on the rapid propagation and quality analysis of in vitro tissue culture plantlets. Few studies have focused on floral bud differentiation. Physiological and molecular mechanisms regulating floral bud differentiation have not been elucidated. Objective: We used paraffin sections and RNA-seq and then analyzed changes in physiological, biochemical, and endogenous hormones to examine floral bud differentiation. Materials and Methods: Buds were harvested in mid-March or early April. After sorting and cleaning, flower buds physiological and biochemical properties were measured and hormones were determined. Morphological observation of S. tonkinensis was carried out using paraffin sections. Results: The transition from the meristematic to the reproductive phase included changes in the shoot apical meristem morphogenesis and cell differentiation. The process includes floral buds initial differentiation and flower primordium differentiation stages; we defined eight stages in total. There was no significant change in soluble content, but soluble protein content gradually decreased. Activities of antioxidant enzymes changed significantly. Total chlorophyll (Chl a + b) content increased significantly. Endogenous hormone levels changed differently during floral bud differentiation. We identified 104,519 expressed genes and 24 were involved in flowering. MADS-box and AP family genes are involved in flower formation and 40 differentially expressed genes associated with floral bud differentiation were identified. Conclusion: The higher soluble sugar, protein, and chlorophyll content and the higher peroxidase activity were beneficial to floral bud differentiation of S. tonkinensis. The dynamic changes in hormone content contribute to differentiating floral buds.
  653 119 -
Propionic acid abrogates the deleterious effects of cerebral ischemic reperfusion injury through nuclear factor-κb signaling in mice
Tiansong Yang, Yulin Wang, Chaoran Li, Yan Yang, Delong Wang, Jing Guo, Tianyu Shi, Yue Wang, Yuanyuan Qu, Qingshuang Wei, Chuwen Feng, Zhongren Sun
Jan-Mar 2020, 16(67):177-182
Background: Cerebral ischemia is caused due to insufficient blood flow to brain cells. This study evaluates the therapeutic effects of propionic acid (PA) on the abnormalities induced during the cerebral ischemic-reperfusion (CIR) injury in mice. Materials and Methods: CIR was induced by complete occlusion of the middle artery of the cerebrum for 15 min and reperfusion for 24 h in the experimental C57BL/6 mice. The analysis of mRNA levels of neuronal nitric oxide synthase (nNOS) was performed by reverse transcriptase polymerase chain reaction (RT-PCR). Inflammatory cytokines such as interleukin-6 (IL-6), IL-1 β, and tumor necrosis factor-alpha were quantitated by enzyme-linked immuno sorbent assay. Apoptosis-related proteins and nuclear factor-κB (NF-κB) were analyzed using Western blot technique. Results: PA demonstrated strong therapeutic effects on controlling the expression of nNOS that was analyzed using RT-PCR. Elevated levels of inflammatory cytokines, caspase-3, caspase-9, and phosphorylated NF-κB during cerebral ischemia were significantly controlled during PA treatment. Conclusion: Our findings demonstrated that PA through oral gavage exhibited healthier effects on the damage caused due to CIR injury.
  662 105 -
Bioactivity-guided fractionation of Cymbopogon caesius (Ness) stapf. revealed isoorientin as an antihypertensive agent

Jan-Mar 2020, 16(67):61-67
Background: Cymbopogon caesius (Ness) Stapf. is a traditional Uyghur medicinal material that is used for the treatment of pain, rheumatism, cold, indigestion, and injury. Objective: The aim of this study was to isolate the antihypertensive activity compounds from C. caesius (Ness) Stapf. Materials and Methods: Cymbopogon caesius (Ness) Stapf. was reflux extracted by EtOH, and the obtained extracts were screened for antihypertensive compounds via the hypertension model of rabbits and rats. The hypertensive extracts were separated via column chromatography and recrystallization and screened for antihypertensive compounds by the hypertension model of rats induced by L-NG-nitroarginine (L-NNA). Their structures were elucidated by nuclear magnetic resonance (NMR) spectroscopy and melt point. Results: Three compounds, namely, isoorientin (Compound I), β-sitosterol (Compound II), and chlorogenic acid (Compound III), were isolated from the dried aerial part of C. caesius (Ness) Stapf. via bioactivity-guided fractionation. Their structures were elucidated by NMR spectroscopy and melt point. Isoorientin may reduce the rats' blood pressure, which was elucidated through the diastolic blood pressure and systolic blood pressure of L-NNA-induced hypertensive rat model experiment. Conclusion: Isoorientin is one of the novel compounds of C. caesius (Ness) Stapf. for the treatment of hypertension diseases.
  617 140 -
Dietary supplementation of Hemidesmus indicus and swimming exercise attenuates oxidative stress in the rat brain

Jan-Mar 2020, 16(67):21-26
Background: In the current investigation, we explored the impact of swimming training and dietary fortification of Hemidesmus indicus (HI) extract on markers of oxidative stress (OS) and antioxidant capacity in the middle-aged Wistar rats. Materials and Methods: Male rats received oral supplementation of HI at a dosage of 50 and 100 mg/kg B.W and swim trained for 30 min/day with 3% intensity, 6 days/week for a duration of 84 days. Results: Training and dietary intake of 100 mg/Kg B.W HI extract enhanced antioxidant enzyme activities in the discrete regions of the brain. The OS indicators were attenuated in response to single or combined interventions. The levels of thiols were significantly upregulated in SW-T (+HI2) group. Conclusion: Our findings suggest that both interventions improve antioxidant status, by attenuating the OS markers, which can be used as a therapeutic strategy in preventing age-associated neurological disorders.
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Cordyceps militaris fraction inhibits angiogenesis of hepatocellular carcinoma in vitro and in vivo
Zhiwei Li, Zhongyi Guo, Jianhua Zhu, Sixue Bi, Yuanyuan Luo, Rongmin Yu, Weijuan Huang, Liyan Song
Jan-Mar 2020, 16(67):169-176
Background: Cordyceps militaris fraction (CMF) was found to inhibit the proliferation of chronic myeloid leukemia K562 cells, oral squamous carcinoma KB cells, and the metastasis of lung cancer cells. This study focuses on the activity of CMF against angiogenesis of hepatocellular carcinoma (HCC). Objectives: The objective of the study is to research the antiangiogenic activity of CMF in HCC cells and the underlying mechanism in vitro and in vivo. Materials and Methods: Transwell migration and invasion assays were used to measure the effects of CMF on migration and invasion of SMMC-7721 cells and human umbilical vein endothelial cells (HUVECs). Tube formation and rat aortic ring assays were used to assess the antiangiogenic potential of CMF. The antiangiogenic mechanism was detected by immunofluorescence and western blot analysis. The nude mice xenografted with SMMC-7721 cells were used to study the antiangiogenic activity of CMF and its underlying mechanism in vivo. Immunohistochemistry analysis was used to evaluate the effect of CMF on the expression of CD31, and western blot analysis was also performed to detect the expression of vascular endothelial growth factor (VEGF) and other related proteins in tumor tissues. Results: CMF inhibited the migration and invasion of SMMC-7721 cells and HUVECs and suppressed VEGF-induced tube formation of HUVECs and the formation of aortic ring capillaries of rats in a concentration-dependent manner. CMF attenuated the phosphorylation of VEGF receptor 2 (VEGFR2), Akt, and ERK in SMMC-7721 cells and HUVECs. CMF significantly inhibited tumor growth in nude mice xenografted with SMMC-7721 cells. CMF reduced the expression level of CD31, western blot analysis indicated that CMF downregulated the expression of VEGFR2 and attenuated the phosphorylation of Akt and ERK in the tumor tissues, which was consistent with the results obtained from in vitro study. Conclusion: CMF can inhibit the angiogenesis of HCC and the mechanism is associated with suppression of the VEGF/VEGFR2 signaling pathway.
  620 115 -
Pharmacokinetic comparisons of six major bioactive components in rats after oral administration of crude and saltwater processed Phellodendri amurensis cortex by ultra-performance liquid chromatography–Mass spectrometry/mass spectrometry
Fan Zhang, Li Meng, Peng-Peng Liu, Guo-Shun Shan, Tian-Zhu Jia
Jan-Mar 2020, 16(67):111-118
Background: The Phellodendri amurensis cortex is a traditional Chinese medicine with multiple pharmacodymic uses (antibacterial, anti-inflammation, antitumor, etc). It is often processed by saltwater to strengthen its effects in terms of nourishing yin to reduce pathogenic fire and reducing asthenic fever. To clarify the principle of saltwater processing, an experiment of pharmacokinetic comparison after oral administration from crude P. amurensis cortex and its saltwater processed product was carried out. Materials and Methods: A validated and sensitive ultra-performance liquid chromatography-mass spectrometry/mass spectrometry (UPLC-MS/MS) method was established for simultaneous quantification of five alkaloids and one triterpene in rat. An UPLC C18column was used for chromatograph separation by an elution program with the mobile phase consisting of 0.1% formic acid and acetonitrile. Each analytes and internal standard, nimodipine, was detected using positive ion scan mode via multiple reaction monitoring mode. All of the validation parameters investigated involving selectivity, precision, accuracy, extraction recovery, matrix effects, and stability shown this approach was suitable to the pharmacokinetic study. Results: Pharmacokinetic profiles showed these parameters of maximum of drug concentration and AUC0-tof alkaloids analytes elevated remarkably after oral administration of P. amurensis cortex processed with the saltwater. Conclusion: The results suggested that the absorption effect from the saltwater processed product was better than those from crude product, which could explain that saltwater processing may enhance the activity of clearing heat and removing toxicity from P. amurensis cortex.
  591 101 -
Comparative study of ultra-high-performance supercritical fluid chromatography and ultra-high-performance liquid chromatography to simultaneous determination of ten components in Radix hedysari
Bo Wang, Xiao-Hua Liu, Zhi-Yuan Xue, Xiu-Yan Yang, Yao-Yao Fang, Shi-Lan Feng
Jan-Mar 2020, 16(67):99-110
Background: In China, dried root of Hedysarum polybotrys Hand-Mazz, known as Hong Qi (Radix hedysari [RH]), is one of the important traditional Chinese medicines (TCMs). The long analysis time was a major limitation when high-throughput techniques are required for research or quality control purposes. Materials and Methods: It is necessary to develop a high?throughput method for the determination of multiple components of TCMs by investigating the separation characteristics of the target with ultra-high-performance liquid chromatography (UHPLC) and ultra-high-performance supercritical fluid chromatography (UHPSFC). Objective: This study intends to establish a high-throughput method for simultaneous determination of ten compounds in RH by UHPSFC and to develop a standardized method to comprehensively evaluate the quality of RH samples. In addition, this study was based on UHPLC and UHPSFC systems to understand and compare the effects of different separation parameters on ten target compounds in RH. A high-throughput and reliable solid-phase extraction method for simultaneous detection of ten target compounds in RH was established and validated. Results: The optimal analysis method was achieved using HSS C18SB column, gradient elution, flow rate of 1.5 mL/min, diode array detection at 270 nm, and back pressure at 12.41 Mpa. The separation method was validated sufficiently by examining the precision, recoveries, linearity, range, limit of detection, and limit of quantification and was successfully applied to quantify the 25 RH samples. In addition, the comparative study also demonstrated the complementary nature of the two separation modes. Conclusion: This is the first study that ten compounds in RH were separated and analyzed simultaneously with UHPSFC-diode array detector. In addition, under the separation conditions of UHPSFC and UHPLC, the comparison of chromatographic parameters shows that a combination of these two separation techniques efficiently separates the ten target compounds in RH. The results obtained in this study are useful for future TCM quality control and high-throughput analysis.
  530 133 -
Study on the inhibition of PLD on IAV-induced pulmonary macrophage based on autophagy and apoptosis
Ling Li, Yi Ning, Ke Wei, Jue Hu, Tao Wu, Cheng Zhao, Fang-Guo Lu, Qing-Hu He
Jan-Mar 2020, 16(67):132-139
Background: Platycodin D (PLD) comes from the main triterpenoid saponins in the dry roots of Platycodon grandiflorum, which has been used in traditional Chinese medicine in the treatment of respiratory disease. This study aimed to evaluate the therapeutic effects of PLD on mouse alveolar macrophages induced by influenza A virus (IAV) in vitro and to investigate the PLD of its action. Materials and Methods: Raw264.7 mouse alveolar macrophages were grown in culture and the Cell Counting Kit-8 assay determined cell viability. Cells were treated for 24 h with increasing doses of PLD (0, 5, 10, 20, 40, 60, 80, 100 μM). The apoptosis was detected by Annexin V-FITC/PI and Tunel assay. Levels of interleukin (IL)-1 β, IL-6 and tumor necrosis factor (TNF)-α of supernatants were assessed using enzyme-linked immunosorbent assay. The numbers of phagophores, autophagosomes, and autolysosomes and the ultrastructure of cells were assessed using transmission electron microscope assay. The relative protein expression of microtubule-associated protein light chain (LC) 3I, LC3II, Beclin-1, and B cell lymphoma (Bcl2) was evaluated by Western blotting. The localizations of Beclin-1 and Bcl2 were viewed with a fluorescence microscope. Results: PLD treatment resulted in significant dose-dependent inhibition of the growth of Raw264.7 cells with the half-maximal inhibitory concentration of 115.74 μM at 24 h. After Raw264.7 cells infected by influenza virus, PLD (2.5, 5, and 10 μM) can improve the activity of cells, reducing apoptosis of cells, decreasing the concentrations of IL-1 β, IL-6, and TNF-α in the supernatant of cells, reducing autophagic injury of cells and regulating the protein expression of Beclin-1 and Bcl2. Conclusion: PLD could inhibit excessive apoptosis and autophagy of macrophage cells induced by IAV in a dose-dependent manner and then inhibits the secretion of inflammatory factors caused by influenza virus infection caused by apoptosis and autophagy and then ultimately regulate the pathogenesis of influenza virus.
  536 121 -
Astragaloside accelerates fracture healing via modulating miR-122/p53 and miR-221/RUNX2 signaling pathways
Shaohui Yang, Lingyan Rong, Hongxia Xu, Huihui Liu, Zhenhai Dong
Jan-Mar 2020, 16(67):119-127
Background: Astragaloside (AS) has been clinically used in the management of fracture, but the underlying mechanism of AS involved in fracture healing is still unknown, so the objective of our study was to explore the above potential mechanism of AS. Materials and Methods: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay, flow cytometry analysis, real-time polymerase chain reaction, Western blot analysis, computational analysis, luciferase assay, and immunohistochemistry (IHC) were utilized to detect the underlying mechanism of AS involved in fracture healing. Results: Cell growth rate, collagen type I, and cell content of osteoblast and MG-63 cell increased while cell apoptosis decreased in a dose-dependent manner compared with controls. MiR-122 and RUNX2 levels showed a stepwise increase while miR-221 and P53 levels showed a stepwise decline as AS concentration increased in osteoblast and MG-63 cells compared with controls. P53 was a virtual target gene of miR-122. Meanwhile, miR-221 directly regulated RUNX2. AS+ group displayed higher messenger RNA (mRNA) levels of miR-122 and RUNX2 than AS− group, while miR-221 and P53 mRNA levels in AS+ group were much lower than AS− group. Results of IHC showed that P53 protein was lowly expressed, while RUNX2 protein was highly expressed in AS+ group compared with AS− group. Conclusion: We identified the effects of the regulatory relationship between AS, miR-122, miR-221, P53, and RUNX2 and their effects on the apoptosis of osteoblasts.
  552 98 -
Ethanol extract of Magnolia sieboldii buds ameliorated esophageal tissue injury induced by gastric acid reflux in rats via regulating the nuclear factor-κB signaling pathway
Li Nan, Hyeon-Hwa Nam, Jin-Cheon Park, Byung-Kil Choo
Jan-Mar 2020, 16(67):161-168
Background: Reflux esophagitis (RE) is one of the gastroesophageal reflux diseases that seriously affect the daily life of patients. Magnolia sieboldii K. Koch has been used as a traditional Chinese herbal medicine for stomach pain relief and deworming and has various physiological activities. Objective: The objective is to investigate the anti-inflammatory and protective effects of the ethanol extract of M. sieboldii buds (MsE) on RE. Materials and Methods: The different concentrations of MsE were used to determine the anti-inflammatory effects on lipopolysaccharide (LPS)-induced cells. In addition, the treatment of MsE on experimental RE rats were used for determining the protective effects of MsE on RE. Results: M. sieboldii treatment effectively inhibited LPS-induced nitric oxide (NO) production, protein expression of inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2), and tumor necrosis factor-alpha (TNF-α) by regulating the activation of nuclear factor-κB (NF-κB) and C-Jun N-terminal kinase/mitogen-activated protein kinase in cells. Furthermore, M. sieboldii treatment significantly ameliorated esophageal tissue damage caused by gastric acid reflux, as well as inhibited the expression of COX-2, TNF-α, and interleukin-1 beta in esophageal tissues through inhibition the activation of NF-κB. Conclusion: We suggested that the ethanol extract of M. sieboldii could consider being an alternative medicine material for treating RE.
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