Pharmacognosy

The role of the herbal medicines, Rehmanniae radix, Citrus unshiu peel, and Poria cocos wolf, in high-fat diet-induced obesity Min Ji Kwon, Jung Nam Kim, Eun Yeong Lim, Yun Tai Kim, Hyungwoo Kim, Myeong Ho Jung, Byung Joo Kim Pharmacognosy Magazine 2019 15(63):363-368 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.

Effects of the herbal medicines on voltage-dependent K+ 2 channels Jeong Nam Kim, Eun Yeong Lim, Yun Tai Kim, Hyungwoo Kim, Byung Joo Kim Pharmacognosy Magazine 2019 15(63):369-377 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.

Research on mechanism of traditional hot processing on velvet antler based on 1H nuclear magnetic resonance and multivariate statistical analysis Jingfeng Li, Hui Sun, Xuefeng Bian, Nanxi Zhang, Yaping Wang, Ying Zong, Hui Zhang, Jiaming Sun Pharmacognosy Magazine 2019 15(63):378-384 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.

The effects of polysaccharides from Rehmannia glutinosa on Caenorhabditis elegans Yanyan Yuan, Nianxin Kang, Yinghui Lu, Xiangzhen Miao, Xiao Zhang, Yonggang Liu, Peng Tan Pharmacognosy Magazine 2019 15(63):385-391 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.

Aroma characteristic analysis of Amomi fructus from different habitats using machine olfactory and gas chromatography-mass spectrometry Huaying Zhou, Dehan Luo, Hamid Gholamhosseini, Zhong Li, Bin Han, Jiafeng He, Shumei Wang Pharmacognosy Magazine 2019 15(63):392-401 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.

Protective effect of curcumin decreases incidence of gastric cancer induced by Helicobacter pylori and N-methyl-N-nitrosourea in rats Duangporn Werawatganon, Kanjana Somanawat, Kawiya Sintara, Somying Tumwasorn, Naruemon Klaikeaw, Prasong Siriviriyakul Pharmacognosy Magazine 2019 15(63):402-409 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.

Rapid identification of medicinal leech by species-specific polymerase chain reaction technology Yang Zheng, Beibei Lu, Yaya Yang, Zhaoqun Jiao, Liqun Chen, Pingtian Yu, Yuping Shen, Huan Yang Pharmacognosy Magazine 2019 15(63):410-415 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.

Pentamethylquercetin inhibited the growth of hepatic ascitic tumor cell H22 by improving metabolic environment and aerobic glycolysis in monosodium glutamate-induced obese mice Wenqi Gao, Xiao Xiao Pharmacognosy Magazine 2019 15(63):416-422 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.

Angiopteris helferiana, a fern with great potential medicinal value: Antiadipogenic, anti-inflammatory, and anti-diabetic activity Ramakanta Lamichhane, Prakash Raj Pandeya, Kyung-Hee Lee, Se-Gun Kim, Dhan Raj Kandel, Hyun-Ju Jung Pharmacognosy Magazine 2019 15(63):423-432 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.

The role of total flavone of Camellia on cerebrovascular vasopasm after subarachnoid hemorrhage in rats Weizhuo Lu, Jiyue Wen Pharmacognosy Magazine 2019 15(63):433-437 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.