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Jul-Sep 2019
Volume 15 | Issue 63
Page Nos. 363-521

Online since Thursday, May 16, 2019

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ORIGINAL ARTICLES  

The role of the herbal medicines, Rehmanniae radix, Citrus unshiu peel, and Poria cocos wolf, in high-fat diet-induced obesity Highly accessed article p. 363
Min Ji Kwon, Jung Nam Kim, Eun Yeong Lim, Yun Tai Kim, Hyungwoo Kim, Myeong Ho Jung, Byung Joo Kim
DOI:10.4103/pm.pm_376_18  
Background: Herbal medicine has been used for the treatment of obesity. Obesity is a very common global health problem, and it is known to be linked to metabolic diseases. Objective: To investigate on the anti-obesity effects of herbal medicines such as Rehmanniae radix (RR), Citrus unshiu peel (CUP), and Poria cocos wolf (PCW) in high-fat diet (HFD)-induced obese mice. Materials and Methods: C57BL/6 mice were fed a normal diet (ND) or an HFD for 6 weeks, and herbal medicines were administered orally three times a week for 8 weeks. Serum biochemical parameters were detected by measuring glucose, triglycerides, and total cholesterol. In addition, histological changes were observed in epididymal adipose tissues. Results: The RR, CUP, and PCW effectively lowered both the body weight and epididymal adipose tissue weight and reduced the adipocyte enlargement. In addition, we examined the serum biochemical profiles of HFD-induced obese mice to confirm the anti-obesity effects of RR, CUP, and PCW. Compared to the ND mice, the administration of RR, CUP, and PCW efficiently reduced the glucose, triglyceride, and total cholesterol serum levels in HFD-induced obese mice. Conclusion: These findings suggested that the herbal medicines, such as RR, CUP, and PCW, are potential novel agents for the prevention and treatment of metabolic diseases, especially obesity.
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Effects of the herbal medicines on voltage-dependent K+ 2 channels Highly accessed article p. 369
Jeong Nam Kim, Eun Yeong Lim, Yun Tai Kim, Hyungwoo Kim, Byung Joo Kim
DOI:10.4103/pm.pm_636_18  
Background: Identification of selective ion channel inhibitors is necessary for understanding the physiological role of these proteins. The voltage-dependent K+ (Kv) channels, Kv2.1 and Kv2.2, are expressed in pancreatic islets, and the development of selective Kv2.1 inhibitors that do not cross-inhibit Kv2.2 may be useful for the treatment of type 2 diabetes. Objective: The aim of this study was to evaluate whether herbal medicines, such as the fruit of Schisandra chinensis (SCF), Atractylodes macrocephala Koidzumi (AMK), Poria cocos Wolf (PCW), Citrus unshiu peel (CUP), Magnolia officinalis Bark (M. officinalis), Alisma canaliculatum (A. canaliculatum), Rehmanniae Radix (RR), and Corni fructus (C. fructus), modulate Kv2 channels and cause insulin secretion. Materials and Methods: We used the whole-cell patch-clamp technique to analyze the effect of these herbal medicines on Kv channels. In addition, human embryonic kidney 293 cells overexpressing Kv2.1 and Kv2.2 channels were used to confirm the role of Kv2 channels. Results: SCF, AMK, PCW, CUP, M. officinalis, A. canaliculatum, and RR inhibited Kv2.1 channel currents in a concentration-dependent manner (100–500 μg/mL). However, C. fructus had no effects on Kv2.1 channel currents. In addition, SCF, AMK, M. officinalis, and A. canaliculatum inhibited Kv2.2 channel currents in a concentration-dependent manner, but PCW, CUP, and RR had no effects on Kv2.2 channel currents. Furthermore, RR, CUP, and PCW increased insulin secretion. Conclusion: These findings suggested that the herbal medicines, RR, CUP, and PCW, are potential novel agents for the prevention and treatment of diabetes.
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Research on mechanism of traditional hot processing on velvet antler based on 1H nuclear magnetic resonance and multivariate statistical analysis p. 378
Jingfeng Li, Hui Sun, Xuefeng Bian, Nanxi Zhang, Yaping Wang, Ying Zong, Hui Zhang, Jiaming Sun
DOI:10.4103/pm.pm_594_18  
Background: The processing of Chinese materia medica is an important part for the preparation of traditional Chinese medicine (TCM) which has been believed that can change the chemical composition and physiological function. Velvet antler is a kind of precious TCM and widely used after hot processing as folk medicines in China. However, no strategy has been presented to reveal the processing principle of velvet antler. Objective: The aim of this study is to develop a method for exploring the mechanism of hot processing on velvet antler. Materials and Methods: In this study, the chemical compositions of fresh velvet antler (FVA) and hot processed velvet antler (HPVA) were compared based on the 1H nuclear magnetic resonance (1H NMR) and multivariate statistical analysis methods to explore the mechanism of hot processing on velvet antler. Results: It showed that hot processing increased phosphorylcholine, taurine, alanine, uridine, phenylalanine, uracil, tyrosine, accompanied by decreased bile acid, choline, lipid 2 and succinic acid in comparison to the FVA, which changed significantly in 52 common metabolites of FVA and HPVA. In addition, a clear separation between FVA and HPVA was obtained by using principal component analysis and 11 “chemical markers” were found by using orthogonal partial least squares discriminant analysis (OPLS-DA) approaches. Particular attention was given to the “chemical markers” with potentially beneficial bioactivities. Conclusion: The study demonstrated that 1H NMR-based chemometric approach could be a promising tool for investigation of the hot processing of velvet antler in a holistic way of TCM.
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The effects of polysaccharides from Rehmannia glutinosa on Caenorhabditis elegans p. 385
Yanyan Yuan, Nianxin Kang, Yinghui Lu, Xiangzhen Miao, Xiao Zhang, Yonggang Liu, Peng Tan
DOI:10.4103/pm.pm_618_18  
Background: Polysaccharides are isolated from Rehmannia glutinosa (RG), a well-known traditional Chinese medicine starting from ancient China. However, their effects on anti-aging activity have not yet been evaluated. Objective: To evaluate the stress resistances and anti-aging effects of polysaccharides from RG by Caenorhabditis elegans N2 wild type. Materials and Methods: Preparing the polysaccharides from Rehmanniae radix preparata (PRRP) and Rehmanniae radix (PRR), we determined the antioxidant activity in vitro, stress resistance, lifespan, fertility, physical growth, locomotion, and impact on the formation of reactive oxygen species (ROS) on C. elegans. Results: The results showed that both polysaccharides have little scavenging ability of free radicals in vitro. After PRRP and PRR treatment, the stress resistance and body bending frequencies of nematodes were significantly increased. PRRP was shown to extend the lifespan and promote physical growth of C. elegans. Both polysaccharides had little effect on fertility and locomotion ability of C. elegans but can reduce excessive intracellular ROS. Conclusion: For C. elegans organism model, both PRRP and PRR are heat resistant and antioxidant; PRRP not only can extend lifespan but can also promote growth and development. PRRP had an anti-aging effect on C. elegans without affecting their reproductive capacity. Based on the scavenging capacity of ROS, we hypothesized that the mechanism of PRRP prolonging lifespan may be related to increase resistance and remove excess free radicals in time.
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Aroma characteristic analysis of Amomi fructus from different habitats using machine olfactory and gas chromatography-mass spectrometry p. 392
Huaying Zhou, Dehan Luo, Hamid Gholamhosseini, Zhong Li, Bin Han, Jiafeng He, Shumei Wang
DOI:10.4103/pm.pm_665_18  
Background: Amomi fructus (AF Lour.) has been used to treat digestive diseases in the context of Traditional Chinese Medicine. Its aroma characteristics have been attracted attention and are considered to be effective markers for determining AF from different habitats. Materials and Methods: In this article, the odor characteristics of AF from three different habitats were investigated and analyzed using gas chromatography-mass spectrometry (GC-MS) and an electronic nose (E-nose). Results: It was found that the E-nose in conjunction with principal component analysis as an analytic tool, showed good performance and achieved a total variance of 93.90% with the first two principal components. A total of 65 aroma constituents among three groups of AF were separated, identified, and calculated using GC-MS. It was observed that the components and the contents were clearly different among the three groups. To confirm the interrelation between aroma constituents and sensors, the contents of 12 aroma ingredients and the response values of six sensors were selected to be trained and tested using the partial least squares. A satisfied quantitative prediction was presented that the contents of selected constituents were accurately predicted by corresponding E-nose sensors with the most determination coefficient of calibration and determination coefficient of prediction of >90%. Conclusion: It was revealed that the E-nose is capable of discriminating AF from different habitats, presenting an accurate, easy-operating, and nondestructive reference approach.
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Protective effect of curcumin decreases incidence of gastric cancer induced by Helicobacter pylori and N-methyl-N-nitrosourea in rats p. 402
Duangporn Werawatganon, Kanjana Somanawat, Kawiya Sintara, Somying Tumwasorn, Naruemon Klaikeaw, Prasong Siriviriyakul
DOI:10.4103/pm.pm_621_18  
Aim: To study the effects of curcumin on Helicobacter pylori and N-methyl-N-nitrosourea (MNU)-induced gastric cancer in rats. Materials and Methods: Male Wistar rats were divided into three groups: control (CO), H. pylori inoculation and 30 ppm MNU in drinking water for 20 weeks (Hp + MNU), and H. pylori and MNU supplemented with 60 mg/kg curcumin for 30 weeks (Hp + MNU + Cur). The stomach was removed to examine nuclear factor kappa B (NF-κB) p65, 8-hydroxy-2'-deoxyguanosine (8-OHdG), cyclin D1, and Ki-67 in gastric epithelial cells by immunohistochemistry. The expression of apoptotic cells was measured by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling reaction and gastric histopathology. Results: Two rats in Hp + MNU developed adenocarcinoma (ADC) of the glandular stomach (40% incidence, n = 5), while in Hp + MNU + Cur, no gastric ADC was found. Histopathology of gastric ADC showed the invasion of malignant cuboidal epithelial cells to submucosal layer. The percentages of NF-κB p65, 8-OHdG, cyclin D1, and Ki-67 immunoreactive cells in Hp + MNU compared with CO were 12.20% ± 1.10% versus 1.86% ± 1.49%, 13.21% ± 0.90% versus 2.84% ± 1.29%, 66.96% ± 5.91% versus 6.06% ± 6.48%, and 42.29% ± 0.08% versus 14.95% ± 0.12%, P < 0.05, respectively. The expression of apoptotic cells significantly increased in Hp + MNU compared with CO (8.41% ± 0.01% vs. 0.53% ± 0.02%, P < 0.05). Curcumin supplementation reduced the gastric cancer incidence compared with Hp + MNU. Percentages of NF-κB p65, 8-OHdG, cyclin D1, and Ki-67 immunoreactive cells in Hp + MNU + Cur compared with Hp + MNU were 4.76% ± 3.73% versus 12.20% ± 1.10%, 1.76% ± 0.94% versus 13.21% ± 0.90%, 24.71% ± 4.62% versus 66.96% ± 5.91%, and 24.99% ± 0.05% versus 42.29% ± 0.08, P < 0.05, respectively. The apoptosis expression was significantly improved (4.14% ± 0.16% vs. 8.41% ± 0.01%, P < 0.05). Conclusion: Curcumin can reduce gastric cancer incidence induced by H. pylori infection and MNU administration through the suppression of key proteins and apoptosis involved in carcinogenesis.
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Rapid identification of medicinal leech by species-specific polymerase chain reaction technology p. 410
Yang Zheng, Beibei Lu, Yaya Yang, Zhaoqun Jiao, Liqun Chen, Pingtian Yu, Yuping Shen, Huan Yang
DOI:10.4103/pm.pm_596_18  
Background: Leech is a well-known animal-derived Chinese medicine; intentionally or unintentionally, interfusing fake products have exposed patients to high risk. Objective: In this study, we would like to find a rapid and accurate technology to identify medicinal leech. Materials and Methods: Three sets of species-specific primer were designed, and polymerase chain reaction (PCR) conditions were optimized for the four species analyzed. Specificity and sensitivity test were carried out, and the reference sample mixtures were analyzed. Finally, the developed method was used to assess the authenticity of commercially available products. Results: Three sets of species-specific primer were confirmed to have high interspecies specificity and good intraspecies stability. The limit of detection was estimated to be 1 ng for all of four assayed species. Subsequent validation demonstrated that the raw material, processed, and even highly processed products of leech can be conveniently authenticated with good sensitivity and precision by this newly established PCR-based method. Conclusion: These designed novel primer sets have shown distinct performance on the identification of adulterate products. The new technology needs no procedure for sequencing and has been successfully employed for the reliable authentication of raw material of leech and even their processed and highly processed products.
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Pentamethylquercetin inhibited the growth of hepatic ascitic tumor cell H22 by improving metabolic environment and aerobic glycolysis in monosodium glutamate-induced obese mice p. 416
Wenqi Gao, Xiao Xiao
DOI:10.4103/pm.pm_605_18  
Aim: We investigated the effects of pentamethylquercetin (PMQ) on the tumor growth in monosodium glutamate (MSG)-induced obese mice. Materials and Methods: At the age of 5 weeks, control and MSG mice were, respectively, divided into five groups (n = 10): Vehicle group; PMQ 5, 10, 20 mg/kg; and metformin (MET) 300 mg/kg groups. All mice were administrated PMQ and MET by gastric gavage from 5- to 24-week age. 22-week-old mice were injection with H22 hepatic ascitic tumor cells. After 2 weeks, animals were anesthetized and blood, tumor, and liver tissues were harvested. Results: Compared with control mice, MSG mice showed obviously metabolic disorders and larger tumor weight and volume than those of control mice. PMQ and MET administration reduced body weight, improved glucose and lipid metabolism, and insulin resistance and inhibited tumor growth in MSG mice. However, PMQ and MET had a litter effect on the tumor growth and metabolic indexes in the control mice. Furthermore, there is significant positive correlation between improved insulin resistance and inhibited tumor growth by chronic PMQ and MET treatment. Further experiments showed PMQ and MET treatment upregulated mRNA expressions of sirtuin 6 (sirt6) both in tumor and liver tissues. Conclusion: Our results demonstrated PMQ decreased tumor growth in the MSG mice and the potential mechanisms might be attributed to upregulated mRNA expressions of sirt6.
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Angiopteris helferiana, a fern with great potential medicinal value: Antiadipogenic, anti-inflammatory, and anti-diabetic activity p. 423
Ramakanta Lamichhane, Prakash Raj Pandeya, Kyung-Hee Lee, Se-Gun Kim, Dhan Raj Kandel, Hyun-Ju Jung
DOI:10.4103/pm.pm_430_18  
Background: Angiopteris helferiana is used as a traditional medicine in some parts of Nepal and South-Asian countries. It is also used in traditional Chinese medicine system. Objective: In this study, we evaluated its anti-oxidant, anti-inflammatory, anti-obesity, and antidiabetic activity in cell and animal mode. Materials and Methods: The rhizome of A. helferiana was extracted with methanol and fractionated to give dichloromethane, ethyl acetate (EA), butanol (BuOH), and water fractions. The residue was again re-extracted with water to give water extract. The anti-oxidant property was measured using 1, 1-diphenyl-2-picryl-hydrazyl assay. The extract and fractions were evaluated for the α-glycosidase inhibition activity. The antiadipogenic and anti-inflammatory activity was studied in 3T3-L1–L1 and RAW 264.7 cells, respectively. The inhibition of expression of adipogenic markers peroxisome proliferator-activated receptor gamma (PPARγ), CCAAT enhancer binding protein alpha (C/EBPα) was studied in 3T3-L1 cells. The anti-obesity and antidiabetic study was done in high-fat diet mice model. Results: The EA, BuOH fraction, and water extract showed good anti-oxidant and α-glycosidase inhibition activity. EA fraction showed good anti-inflammatory activity. EA and BuOH fraction showed good antiadipogenic activity, reducing significantly, the lipid and adipogenic markers (PPARγ, C/EBPα) in 3T3-L1 cells. The BuOH fraction at 300 mg/kg/day dose showed good anti-obesity and antidiabetic activity in the in-vivo study after the evaluation of body weight, blood lipid, blood glucose, and lipid accumulation in adipose tissue and liver. Conclusion: Overall, A. helferiana is a good source of anti-oxidative, anti-inflammatory, anti-obesity, and antidiabetic component.
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The role of total flavone of Camellia on cerebrovascular vasopasm after subarachnoid hemorrhage in rats p. 433
Weizhuo Lu, Jiyue Wen
DOI:10.4103/pm.pm_593_18  
Background: This study was undertaken to explore the role of total flavones of Camellia (TFC) on the cerebrovascular dysfunction in rats after experimental subarachnoid hemorrhage (SAH). Materials and Methods: The contraction and dilation of cerebral basilar artery (BA), nitric oxide (NO) level, and nitric oxide synthase (NOS) activity in rat serum and the expression of endothelial NOS (eNOS) in cerebral vessels were measured. Furthermore, the effect of potassium channel blockers, endothelium removal, L-NG-nitroarginine methyl ester (L-NAME), or prostacyclin I2 (PG I2) production inhibitor on the response of isolated BA derived from normal rats was also evaluated to explore the underlying mechanism of vasodilation induced by total flavones. Results: The contraction of rat BA to U46619 markedly increased and the vasodilation to acetylcholine remarkably reduced after SAH. Interestingly, these vascular dysfunctions were profoundly ameliorated by pretreatment of TFC. Moreover, total flavones could induce a concentration-dependent relaxation in isolated BA from normal rats, which was obviously eliminated by co-application of potassium channel blockers, ChTx and Apamin, application of L-NAME, or endothelial removal. In addition, total flavones pretreatment obviously improved the expression of eNOS in BA, serum NO level, and NOS activity at 48 h after SAH. Conclusion: These findings revealed that TFC has protective effect on cerebrovascular dysfunction after SAH and demonstrated that the protection could be due to its upregulation of eNOS expression and activation of potassium channel.
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Cytotoxicity, protein kinase inhibitory activity, and docking studies of secondary metabolites isolated from Brownea grandiceps Jacq p. 438
Ehab M Mostafa, Arafa Musa, Mohamed A Abdelgawad, Ehab A Ragab
DOI:10.4103/pm.pm_35_19  
Background: Numerous kinases are excessively secreted in cancer. Consequently, inhibition of kinase enzymes has a basic role in the treatment of cancer through suppression or prevention of cancer cell multiplication. Due to its kinase inhibitory activity, flavonoids are expected to be of great importance in the discovery of new anticancer drugs. Based on the chemotaxonomic relationship among the plant genera, Brownea grandiceps is expected to be rich in flavonoids. Objective: The objective was to study the cytotoxicity, kinase inhibitory activity, and docking of the isolated metabolites. Materials and Methods: Ultraviolet, nuclear magnetic resonance, and mass spectroscopy were used for the identification and confirmation of the active metabolites. Ethyl acetate extract of B. grandiceps was chromatographed. Cytotoxicity (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide), protein kinase (PK) inhibitory assays, and molecular operating environment (using Molecular Operating Environment [MOE], 2016.08) were performed. Results: Catechin (1), quercitrin (2), trans-taxifolin-3-O-α-L-rhamnopyranoside (3), 13R-hydroxy-9Z, 11E-octadecadienoic acid (4), and 13R-hydroxy-9Z, 11E-, 15Z-octadecatrienoic acid (5) were isolated and identified. Quercitrin (2) exhibited the highest cytotoxicity against MCF-7 (inhibitory concentration for 50% [IC50]: 4.24 μM) and moderate cytotoxicity against HepG2 cell (IC50: 29.58 μM). Others showed moderate cytotoxicity; compounds 1–3 were tested against Aurora B, CDK4/cyclin D1, COT, IGF1-R, and FAK PKs, where quercitrin showed the highest inhibitory activity against Aurora B and CDK4/Cyclin D1 (IC50: 4.78 and 3.22 μM). Docking of quercitrin against CDK4/Cyclin D1 confirmed its cytotoxic profile. Conclusion: The metabolites were first isolated from B. grandiceps. The mechanism of action against kinases enzyme was established and confirmed by docking studies of quercitrin at CDK4/Cyclin D1 using MOE program.
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Antitumor and immunoregulation effects and mechanism of n-butanol fraction from Zanthoxylum avicennae in H22 mice p. 443
Hong-Xin Cui, Fang-Rong Cheng, Nan-Nan Zheng, Shan-Shan Jia, Ke Yuan
DOI:10.4103/pm.pm_251_18  
Background: The aim of this study was to study the antitumor and immunomodulatory effects and its mechanism of the n-butyl alcohol extract of the ethanol extracts in Zanthoxylum avicennae (Lam.) DC. for the mice with liver cancer H22. Materials and Methods: To inoculate H22 tumor plant to the mice and establish three different animal models of liver cancer solid tumor ascitic tumor and immunodeficiency in mice H22 and then divide the mice into six groups: blank group, model group, positive group (cyclophosphamide or astragalus polysaccharide), drug dosages group of high, medium, and lower (1.28 g/kg, 0.64 g/kg, 0.32 g/kg). After 2 weeks of intragastric administration, the content of aspartate transaminase (AST), alanine transaminase (ALT), tumor necrosis factor-α (TNF-α), and interleukin-2 (IL-2) in the blood serum of the mice with solid cancer and the level of superoxide dismutase (SOD), malondialdehyde (MDA), TNF-α, and IL-2 in the blood serum of the immunodeficiency mice were determined. At the same time, the indexes of liver, spleen, and thymus gland organs in solid tumor mice and immunodeficiency mice were determined, the life elongation rate of the mice with liver cancer H22 ascitic tumor was observed, and then the changing status of the tumor tissue by the method of HE dyeing was observed. Results: The contents of TNF-α and IL-2 in the serum of the high-dose group of the solid tumor model were obviously higher than the normal group, with very significant difference (P < 0.01) and very high tumor inhibition rate of 54.61%, and the contents of transaminase AST, ALT, and the tumor inflammatory factors such as TNF-α and IL-2 in the mice serum were obviously decreased, and the tumor weights were reduced greatly; the contents of MDA, TNF-α, and IL-2 in the serum of the mice of the immunodeficiency group were obviously decreased, with obvious increase of the SOD activity and the organ indexes of the liver, spleen, and thymus gland; the life extension rate of the mice in the high-dose group of the ascitic tumor model was obviously increased. Conclusion: The n-butyl alcohol extract of Z. avicennae improved the survival quality of the H22 tumor-bearing mice and enhanced their immune ability to exhibit very excellent antitumor activity through improving the inflammatory factors of the tumor-bearing mice. The mechanism may be that it had the function of antitumor just by improving the immune organs' quality of the tumor-bearing mice, and at the same time increased the secretion of the cell inflammatory factors such as TNF-α and IL-2 and thus strengthened the immunocompetence and improved the lipid peroxidation in the bodies of tumor-bearing mice.
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Determination of trace metal concentrations in different parts of the khat varieties (Catha edulis) using inductively coupled plasma-mass spectroscopy technique and their human exposure assessment p. 449
Mohammed Al Bratty, Waquar Ahsan, Hassan A Alhazmi, Ibraheem M Attafi, Ibrahim A Khardali, Siddig I Abdelwahab
DOI:10.4103/pm.pm_658_18  
Background: Khat (Catha edulis, family: Celastraceae) is a plant that is native to Africa and Arab peninsula and is used for their amphetamine-like properties. Although the use of Khat is banned in Saudi Arabia, people particularly in southern Jazan province manage to get it from Yemen, and the use is increasing steadily. Objective: Five most commonly used varieties of Khat namely Gaifi, Kofat, Jahasha, Faqarah Menjed, and Faqarah Aswad were selected for the study. Materials and Methods: Metal ion concentrations were determined using inductively coupled plasma-mass spectroscopy. Since Khat is available as one bundle consisting of three parts of the plant, metal ions in all three parts were determined separately for comparison purpose. The concentrations (mg/kg) of 20 metal ions were determined in Nwaif leaves (new and smaller in size), Gafra leaves (old and larger in size), and stem of the plant and compared with the Provisional Tolerable Weekly Intake (PTWI) and acceptable daily intake (ADI) of metal ions to study the health hazards posed by them. Results: The non-essential metal ion Strontium (Sr) was present in highest abundance in all the samples with a concentration range of 498.6 ± 18.9–3837 ± 52.1 mg/kg followed by Copper (215.4 ± 12.3–3054 ± 45.2 mg/kg), Zinc (23.17 ± 0.4–1490 ± 32.6 mg/kg), and Manganese (108 ± 5.8–1357 ± 18.6 mg/kg). Several toxic heavy metal ions including Arsenic, Lead, and Cadmium were also present in trace concentrations in many samples. Conclusion: Many metal ions were observed to be present in concentrations much higher than their PTWI and ADI which further allude to the extremely hazardous nature of Khat plants. Multicomponent variate analyses were also performed using chemometric methods to establish the possible correlation between samples.
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Chinese native medicinal plant Euphorbiaceae show antitumor and anti-oxidant features in lewis lung cancer-bearing mice p. 459
Xiao-Ming Zhao, Li-Yuan Qu, Zhonghai Yan, Yun Yang, Ying Ma, Zhao-Lun Dong, Zhen-Ming Li, Meng-Wen Li, Xin Wang, Fei Jiao
DOI:10.4103/pm.pm_540_18  
Background: As a promising means, natural products from Chinese herb provide valuable sources for cancer therapy. Euphorbiaceae has been used as a remedy for the treatment of many diseases, including cancer. However, studies about its effects on lung cancer are limited, and the mechanisms are not well established. Objective: The present study aims to investigate the anticancer effects of Euphorbiaceae extract (EE) in vitro and in vivo, as well as the potential mechanisms. Materials and Methods: In the present study, 3-(4,5-dimethythiazol-. 2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay was performed to investigate the effects of EE on Lewis lung adenocarcinoma cell (LLC) viability. Flow cytometric analysis was used to observe the changes of apoptosis and cell cycle of large cell carcinoma (LCC). The gene expressions were detected by real-time reverse transcription–polymerase chain reaction and Western blot. The tumorigenicity assay was performed to evaluate the inhibitory effects of EE in vivo. The activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) were also determined. Results: MTT assay demonstrated that EE significantly inhibited the growth of LLC in a dose-dependent manner in vitro. Flow cytometric analysis revealed that EE markedly induced apoptosis and G0/G1 arrest of the LCC cells. Furthermore, we found that EE led to the expression changes of apoptosis-related genes, with the decrease of Bcl-2 and the increase of Bax and caspase-9. The results from tumor-bearing animals further confirmed that the administration of EE significantly suppressed the tumor growth in vivo. Meanwhile, the activities of serum SOD, CAT, and GSH-Px increased significantly after the treatment of EE. Conclusion: These results suggested that the anticancer effects of EE may be involved in apoptosis induction, proliferation suppression, and oxidant scavenging.
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Effect of frost on flavonol glycosides accumulation and antioxidant activities of mulberry (Morus alba L.) leaves p. 466
Yu Xiaofeng, Zhao Shuang, Zhao Li, Wang Dan, Fan Xiaoman, Ouyang Zhen
DOI:10.4103/pm.pm_493_18  
Background: Both Pharmacopoeia of the People's Republic of China and the ancient Chinese herbal formulas recorded that mulberry leaves collected after frost had good quality. However, the reason has not yet been fully elucidated. Objective: We investigated the effect of frost on the accumulation of flavonoids and antioxidant activities of mulberry leaves. Materials and Methods: Liquid chromatography-mass spectrometry and high-performance liquid chromatography were used to analyze chemical components and determine the content of five flavonol glycosides from mulberry leaves collected before and after frost, respectively. Antioxidant activities of the same mulberry leaves were evaluated by total antioxidant capacity (TAC), Fe2+ equivalent (FeE), reducing power (RP), 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging assay, and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid (ABTS) free radical scavenging assay. Results: Ten compounds were identified as flavonol glycosides exception of chlorogenic acid. Quantitative analysis showed that content of isoquercitrin, astragalin, and kaempferol-3-O-(6''-acetyl)-β-D-glucopyranoside reached a maximum of 3.05 mg/g, 0.70 mg/g, and 0.69 mg/g after Frost's Descent, respectively. Moreover, the lowest value of flavonol glycosides appeared in August. The antioxidant activities were also found to have the same tendency. The maximum value of TAC, FeE, RP, DPPH and ABTS were 64.3 rutin equivalent (RE) mg/g, 46.2 RE mg/g, 31.3 RE mg/g, 22.5 RE mg/g and 26.7 RE mg/g, respectively, in November. They were 1.4 times, 1.4 times, 1.6 times, 1.6 times, and 1.9 times of the minimum values, respectively, in August. There was a significantly and positively correlation between antioxidant activities and content of flavonol glycosides (P < 0.01). Conclusion: Frost is beneficial to the accumulation of flavonol glycosides and the improvement of antioxidant activities of mulberry leaves.
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Headspace–solid-phase microextraction gas chromatography method to quantify Thymus vulgaris essential oil in polymeric nanoparticles p. 473
Lorena Lugo-Estrada, Sergio Arturo Galindo-Rodríguez, Luis Alejandro Pérez-López, Noemí Waksman de Torres, Rocío Álvarez-Román
DOI:10.4103/pm.pm_277_18  
Background: Thymus vulgaris essential oil (Tv-EO) is known to have antibacterial, antifungal, and antioxidant activities. Encapsulation of Tv-EO in polymeric nanoparticles (NPs) can prevent volatilization of its components and can provide protection against external agents. Under these circumstances, it is crucial to assure the presence and quantity of the Tv-EO components (γ-terpinene, thymol, and carvacrol) in the NPs. Objective: To determine the chemical composition and physicochemical characterization of Tv-EO as well as develop and validate a HSPM-gas chromatography (GC) method for the analysis of Tv-EO components encapsulated in NPs. Materials and Methods: Tv-EO was characterized by physicochemical analysis for relative density, refractive index, and optical rotation and analyzed by GC flame ionization detector and GC-mass spectrometry. The headspace–solid-phase microextraction-gas chromatography (HS-SPME-CG) validation was assessed, Tv-EO-NPs were prepared by nanoprecipitation, and its properties were determined by photon correlation spectroscopy. Results: Tv-EO was characterized by physicochemical analysis for relative density (0.934 g/cm3), refraction index (1.559), and optical rotation (−0.084°). Seventeen components were identified in Tv-EO; among these, the sesquiterpenes, thymol (34.28%), o-cymene (31.78%) and γ-terpinene (13.22%). The method was validated for linearity (R2 ≥ 0.99), precision (intraday 7.02, 10.33, and 8.60 and inter-day 10.60, 10.60, and 10.99), accuracy (99.35, 109.4, and 98.84%) and robustness for γ-terpinene, thymol and carvacrol, respectively. The limit of detection and limit of quantification were calculated as 0.69, 0.40, and 0.39 μg/mL and 2.11, 1.22, and 1.20 μg/mL for γ-terpinene, thymol, and carvacrol, respectively. An encapsulation percentage of 47.51% of total essential oil was obtained. Conclusion: The experimental data show that HS-SPME reduces interference of the NP-matrix and concentrates the Tv-EO components. HS-SPME-CG can be considered as a good alternative to the already existing methods for analysis of essential oil encapsulated in NPs.
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Rapid and simultaneous analysis of five free anthraquinone contents in rhubarb during the stir-frying with rice wine process by near infrared reflectance spectroscopy p. 479
Xingyi Chen, Jintao Xue, Weiying Wang, Bihua Ma, Guo Huang, Yu Chen, Liming Ye
DOI:10.4103/pm.pm_562_18  
Background: The active ingredients of Traditional Chinese Medicines (TCMs) vary greatly with the degree of stir-frying; so, rapid analysis of the active content is very important for the processing of TCMs. Objective: In this study, near infrared reflectance (NIR) spectroscopy was used to develop a new method for the rapid online analysis of five free anthraquinones (aloe-emodin, rhein, emodin, chrysophanol, and physcion) during the stir-frying process for rhubarb. Materials and Methods: With partial least-squares (PLSs) and artificial neural networks (ANN) regression, calibration models were generated based on five free anthraquinone contents, as measured by high-performance liquid chromatography. Results: The results indicated that the 2 types of models were robust, accurate, and repeatable for five free anthraquinones. Moreover, PLS as the linear model was more suitable for developing the NIR models of the five free anthraquinones than ANN. The performance of the optimal models was achieved as follows: the coefficient of determination for prediction (R2pre) for aloe-emodin, rhein, emodin, chrysophanol, and physcion was 0.9161, 0.9699, 0.9655, 0.9611, and 0.9724, respectively; the root mean square error of prediction was 0.0251, 0.0445, 0.3333, 0.0862, and 0.0211, respectively. Conclusion: The established NIR models could apply to determine the content of five free anthraquinones in rhubarb. This work demonstrated that NIR may be an effective online analysis method to reflect the quality of TCM industrial manufacturing processes.
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(-)-Epigallocatechin-3-gallate protects against uric acid-induced endothelial dysfunction in human umbilical vein endothelial cells p. 487
Hua Xie, Jianqin Sun, Yanqiu Chen, Min Zong, Danfeng Xu, Yan Wang
DOI:10.4103/pm.pm_659_18  
Background: Hyperuricemia is recently reported to be associated with hypertension, metabolic syndrome, and vascular damage. (-)-Epigallocatechin gallate (EGCG) is a major polyphenol component of green tea involving in potent anti-inflammatory and antioxidant effects. The study assessed the effect of EGCG on uric acid (UA)-induced vascular damage. Materials and Methods: The cell viability and angiogenic formation of human umbilical vein endothelial cells (HUVECs) treated with UA and EGCG were determined by methylthiazol tetrazolium and tube formation assays, respectively. The expression level of inflammatory cytokine and vascular factor, including nuclear factor-kappa B (NF-κB), monocyte chemotactic protein-1 (MCP-1), intercellular adhesion molecule 1 (ICAM-1), cycloxygenase-2, tumor necrosis factor (TNF-α), induced nitric oxide synthetase (iNOS), endothelin-1 (ET-1), and von Willebrand factor, were examined by real-time quantitative reverse transcription polymerase chain reaction. The expression of P65 in HUVECs treated with UA in different time points or different concentrations was detected by Western blotting. Results: EGCG suppressed UA-inducing HUVEC cells death. UA treatment of HUVEC significantly increases the expression of P65. EGCG significantly inhibited the UA-induced mRNA expression of NF-κB, MCP-1, ICAM-1, TNF-α, iNOS, and ET-1 in HUVECs. Functional analysis of angiogenic inhibition showed that pretreatment with EGCG improved the vascular tube formation. Conclusion: Our results suggested that UA-induced inflammatory cytokine production and impaired endothelial cell function in HUVECs were attenuated by EGCG. These data indicated that EGCG has a therapeutic potential for UA-mediated endothelial damage.
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Anticancer potential of seed extract and pure compound from Phoenix dactylifera on human cancer cell lines p. 494
Ebtesam S Al-Sheddi
DOI:10.4103/pm.pm_623_18  
Background: Phoenix dactylifera (Palmacea), known as date palm is a widespread economical plant in the Middle Eastern. The dietary fiber in P. dactylifera seeds has important therapeutic use in medical condition such as diabetes, obesity, hypertension, colorectal, and prostate cancers. Objectives: The objective is to isolate, characterize the major bioactive components and evaluate the cytotoxic activity of extract and isolated pure compound of P. dactylifera. Materials and Methods: P. dactylifera extract (DE) was obtained by maceration. The pure compound, identified as oleic acid (OA) was isolated by column chromatography. Cytotoxicity assessment was done by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide assay and morphological alterations in HepG2, A-549, and MCF-7 cells and bioactive compounds were evaluated by gas chromatography/mass spectrometry. Results: The DE showed a dose-dependent cytotoxicity in all the testes cell lines. The cell viability at doses of 250, 500, and 1000 μg/ml of DE was found as 87%, 75%, and 48% in HepG2; 95%, 85%, and 78% in A-549; and 77%, 51%, and 35% in MCF-7 cells, respectively. The GCMS analysis indicated the presence of 37 compounds. The fatty acids and esters, fatty alcohols, and steroid ester were predominant in the DE. The IC50 value of isolated pure compound (OA) was determined at 735.2 μg/ml in HepG2, 909.1 μg/ml in A549, and 675.6 μg/ml in MCF-7 cells. Conclusion: These results suggest that DE has promising anticancer potential and OA could be the compound contributing to cytotoxicity.
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Polysaccharides from radix astragali exert immunostimulatory effects to attenuate the dampness stagnancy due to spleen deficiency syndrome p. 500
Wenxiao Zhao, Lijun Chen, Ning Cui, Xuming Ji, Haiqiang Jiang, Tiantian Deng, Shijun Wang
DOI:10.4103/pm.pm_503_18  
Background: Radix Astragali is an herb with tonifying qi or adaptogenic effects and commonly used for spleen-qi deficiency in traditional Chinese medicine. Astragalus polysaccharides (APS) is one of the major bioactive ingredients, and little is known on APS in attenuating the dampness stagnancy due to spleen deficiency (DSSD). Objective: The objective of the study is to investigate the potential mechanism of action of APS underlying attenuating water and fluid retention of rats with the DSSD syndrome in order to provide novel insights into the application of APS on modulating the DSSD syndrome. Materials and Methods: Four-week-old Wistar rats were fed with one high-fat and low-protein diet and subjected to exhaustive swimming to induce the DSSD model for 8 weeks. APS (300, 600, and 1200 mg/kg) and Shen Ling Bai Zhu San (SLBZS) (2.5 g/kg) were administrated starting at 9 weeks for 2 weeks. Results: APS increased the body weight, decreased the water load index, and attenuated symptoms of the DSSD syndrome. APS also increased spleen and thymus indices, the percentage of CD3+, the percentage of CD3+ CD4+, and CD3+ CD4+ to CD3+ CD8+ ratio and decreased levels of serum interleukin (IL)-6, IL-10, and tumor necrosis factor-α in DSSD rats. Significant effects were seen for toll-like receptor 4 (TLR4) at both protein and gene expression levels with amelioration of expression levels in APS groups. Conclusion: These findings suggest that APS may be used to attenuate the fluid retention of the DSSD syndrome through the immunoregulatory effect, which is driven in part by the modulation of TLR4.
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Inhibition of oxidative stress and autophagy by arbutin in lipopolysaccharide-induced myocardial injury p. 507
Beibei Zhang, Mengnan Zeng, Benke Li, Yangyang Wang, Yuxuan Kan, Shengchao Wang, Yijia Meng, Jiji Gao, Weisheng Feng, Xiaoke Zheng
DOI:10.4103/pm.pm_552_18  
Background: Sepsis is a syndrome characterized by a systemic inflammatory response. Arbutin (Ar) is an active natural product known for its bactericidal and anti-inflammatory effects. Many studies have reported the diverse pharmacological actions of Ar, but there is no relevant research on the effect of Ar on lipopolysaccharide (LPS)-induced myocardial injury. Objective: The purpose of this study was to investigate the effect of Ar on LPS-induced myocardial injury and its underlying mechanisms. Materials and Methods: The levels of tumor necrosis factor alpha, interleukin 6, cardiac troponin-I, and procalcitonin were detected by ELISA. The levels of phosphorylated c-Jun N-terminal kinase (p-JNK), phosphorylated extracellular regulated protein kinase (p-ERK), and phosphorylated p38 (p-p38) proteins were detected by flow cytometry using Cytometric Bead Array. Western blot was used to detect the expression of autophagy-related and estrogen receptor (ER)-associated proteins. Levels of the oxidative stress-related markers were detected by the cuvette assay. Results: The levels of the inflammatory factors, LC3B, malondialdehyde, p-JNK, and p-p38 were increased in LPS-treated rats, while the ERK, total superoxide dismutase, glutathione peroxidase, p62, and ER-associated proteins were decreased. These effects could be effectively reversed by Ar, which could be blocked by ER antagonist ICI182780. Our previous study found Ar to possess an estrogen-like activity. Conclusion: Ar inhibits the oxidative stress and autophagy and offers protection from the LPS-induced myocardial injury via the ER pathway.
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Heat stress resistance effect of flavonoids from Penthorum chinense Pursh on Caenorhabditis elegans p. 514
Shiling Feng, Liang Fu, Yujie Wang, Handong Wang, Ming Yuan, Yan Huang, Hongyu Yang, Chunbang Ding
DOI:10.4103/pm.pm_500_18  
Background: Excessive reactive oxygen species (ROS) may overwhelm antioxidative defense, which is implicated in the onset and progression of various diseases. Flavonoids extracted from Penthorum chinense Pursh have a remarkable spectrum of bioactivities, while these pharmacological activities of flavonoids are at least partially due to their scavenging ROS and antioxidant ability. Therefore, we used the multicellular Caenorhabditis elegans as a model animal to investigate their antioxidant property and the possible molecular mechanisms in this study. Materials and Methods: The chemical compositions in P. chinense flavonoids were identified by ultra-performance liquid chromatography/time-of-flight mass spectrometry. Their anticancer and antioxidant ability in vitro were evaluated by inhibitory rate of HeLa cells, free radical scavenging, and iron-chelating ability. In vivo, synchronized L4 larvae C. elegans were treated by P. chinense flavonoids for 48 h. The survival rates of C. elegans were measured under heat stress condition. The antioxidant effect of P. chinense flavonoids on C. elegans was assessed by superoxide dismutase (SOD) and catalase (CAT) activities and malondialdehyde content. Results: P. chinense flavonoids was composed of six quercetin derivatives, two kaempferol derivatives, six pinocembrin derivatives, thonningianins A, and vanillic acid glucoside. P. chinense flavonoids not only showed a significantly inhibitory rate of HeLa cells but also exerted iron chelation and free radical scavenging ability. Furthermore, P. chinense flavonoids could extend the mean lifespan of C. elegans by approximately 17% under heat stress, which might be due to the increase of SOD and CAT activities. Conclusion: The study demonstrated that P. chinense flavonoids might be developed as a promising natural agent against environmental stress.
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