Lignans are the main active components of Schisandrae Chinensis Fructus for liver disease treatment:a review

As a traditional Chinese herbal medicine,Schisandrae Chinensis Fructus(SC)has been used in medicine and food industry due to its health care and therapeutic effects.Over the past 20 years,the use of SC and its active ingredient lignans in the prevention and treatment of liver diseases has been increasing,and their hepatoprotective effects has increased the interest of the public and academia.Therefore,in the present work,we first determined the effectiveness of SC in the treatment of liver diseases such as metabolic associated fatty liver disease,alcoholic liver disease,cholestatic liver disease and acute liver injury.Subsequently,the pharmacological effects and molecular mechanisms of lignans,the active components of SC,for liver disease treatment were comprehensively summarized for the first time.The results showed that the lignans in SC could achieve hepatoprotective effects by regulating lipid metabolism,anti-fibrosis,anti-inflammation,anti-oxidation,anti-tumor and regulating bile acid metabolism.The mechanism mainly involved adenosine 5’-monophosphate-activated protein kinase,endoplasmic reticulum stress,sterol regulatory element binding protein 1c,autophagy,transforming growth factor-β,mitogen-activated protein kinase,microRNA,nuclear factor kappa-B,nuclear factor erythroid-2-related factor 2,heat shock proteins and pregnane X receptor signaling pathways.These results can lay a scientific foundation for the development of hepatoprotective drugs or functional foods from SC/lignans.

Effects of Different Decolorants on Retention Rate of Total Triterpenes in Fruit and Rattan Stems of Schisandra chinensis(Turcz.) Baill

This study was to investigate the decoloration effects of activated clay, activated carbon, diatomite, Ca group bentonite, kaolin, activated aluminum oxide, magnesium oxide and attapulgite. Ultraviolet-visible spectroscopywas adopted to de- tect the absorbanees of the extracts of Schisandra chinensis fruit and rattan stems, the contents of total triterpenoids in S. chinensis fruit and rattan stems were deter- mined, and the decoloration rate and the retention rate of total triterpenoids of S. chi- nensis fruit and rattan stems were calculated. The results indicated that, attapulgite exhibited the best decoloration effect on the extract of S. chinensis fruit, with a de- coloration rate up to 60.47%, activated carbon exhibited a better decoloration effect on the extract of S. chinensis rattan stems, with a decoloration rate up to 69.24%, and they had relatively higher retention rates of total triterpenoids. In the eight de- colorants, attapulgite showed the best decoloration effect on the extract of S. chi- nensis fruit, and activated carbon exhibited the best decoloration effect on the ex- tract of S. chinensisrattan stems.

胚乳和外源赤霉素对五味子[Schisandra chinensis (Turcz.) Baill]种子发芽的影响

以组织培养和土壤播种的未经层积处理的五味子种子为材料，检测胚乳和外源赤霉素（GA3）对发芽影响的结果表明，在不加生长调节物质的MS培养基上，去掉外种皮带全胚乳的种子发芽率只有2．7％，而带1／2和1／3胚乳的种子发芽率则分别为46％和70．6％；培养基中添加GA，，可提高去掉外种皮带全胚乳和带1／2胚乳种子的发芽率，而对带1／3胚乳的种子则无效果。土壤播种时，用10—1000mg·L—GA3对去掉外种皮种子进行1~10d前处理，其中100mg·L—GA3前处理10d的发芽率最高，达到38．7％。

Nortriterpenoids from the fruit stalk of Schisandra chinensis (Turcz.) Baill.

Objective:The fruit stalk of Schisandra chinensis(Turcz.)Baill.(S.chinensis)has been found to contain bioactive components similar to the fruit of S.chinensis.Here,we report a recent discovery about new nortriterpenoids with a novel skeleton and anti-gastric cancer activity,which were isolated from the fruit stalk of S.chinensis.Methods:The chemical components of ethyl acetate extract from 70%ethanol extract from S.chinensis fruit stalk were separated,purified,and identified by liquid chromatography methods(silica gel,ODS,HPLC)and extensive spectroscopic analyses(NMR,IR,UV,MS,CD).Results:Two new nortriterpenoids,schilancitrilactone M and 25-hydroxyl schindilactone D(1 and 2),along with ten known nortriterpenoids(3-12)were isolated from the fruit stalk of S.chinensis.The isolated compounds were tested for their cytotoxic activities against MGC-803 cells,and the results showed that compounds 6-8 possessed significant activities with IC50 of 9.01,11.77,and 2.74μmol/L,respectively.Conclusion:Twelve nortriterpenoids including two new compounds were isolated from the fruit stalk of S.chinensis for the first time.Among them,compounds 6-8 showed significant anti-gastric cancer activities.We postulated that the fruit stalk of S.chinensis could be used as an anti-gastric cancer drug.

Schisandra chinensis (Turcz.) Baill

Schisandra chinensis (Turcz.) Baill. is a deciduous woody vine, and belongs to the Schisandraceae family. This species is a horticultural plant with edible fruit. It is used medicinally and for making rope. It is distributed in Hebei, Heilongjiang, Jilin, Liaoning, Nei Mongol, Shanxi

Effect of Schisandra chinensis polysaccharide on intracerebral acetylcholinesterase and monoamine neurotransmitters in a D-galactose-induced aging brain mouse model

BACKGROUND： The most prominent characteristic of brain aging is decreased learning and memory ability. The functions of learning and memory are closely related to intracerebral acetylcholinesterase （ACHE） and monoamine neurotransmitter activity. Previous studies have shown that Schisandra chinensis polysaccharide has an anti-aging effect. OBJECTIVE： To explore the effects of Schisandra chinensis polysaccharide on AChE activity and monoamine neurotransmitter content, as well as learning and memory ability in a D-galactose-induced aging mouse brain model compared with the positive control drug Kangnaoling. DESIGN, TIME AND SETTING： Completely randomized, controlled experiment based on neurobiochemistry was performed at the Pharmacological Laboratory, Henan University of Traditional Chinese Medicine from September to December 2003. MATERIALS： Schisandra chinensis was purchased from Henan Provincial Medicinal Company. Schisandra chinensis polysaccharide was obtained by water extraction and alcohol precipitation. Kangnaoling pellets were provided by Liaoning Tianlong Pharmaceutical （batch No. 20030804; state drug permit No. H21023095）. A total of 50 six-week-old Kunming mice were randomly divided into five groups： blank control, model, Kangnaoling, high and low dosage Schisandra chinensis polysaccharide groups, with 10 mice per group. METHODS： Mice in the blank control group were subcutaneously injected with 0.5 mL/20 g normal saline into the nape of the neck each day, while the remaining mice were subcutaneously injected with 5% D-galactose saline solution （0.5 mL/20 g） in the nape for 40 days to induce a brain aging model. On day 11, mice in the high and low dosage Schisandra chinensis polysaccharide groups were intragastrically infused with 20 mg/mL and 10 mg/mL Schisandra chinensis polysaccharide solution （0.2 mL/10 g）, respectively. Mice from the Kangnaoling group were intragastrically infused with 35 mg/mL Kangnaoling suspension （0.2 mL/10 g）, and the mice in the model group were intragastrically infused with the same volume of normal saline （0.2 mL/10 g） once per day for 30 consecutive days. MAIN OUTCOME MEASURES： Two hours after the final administration, pathohistological changes in the cerebral cortex and hippocampus were observed using hematoxylin ＆ eosin staining. AChE activity was detected using chromatometry. Monoamine neurotransmitter content was measured using fluorimetry. Learning and memory was measured using the step down test and darkness avoidance test. RESULTS： Both Schisandra chinensis polysaccharide and Kangnaoling improved pathological injury to the cerebral cortex and hippocampus in a mouse model of brain aging. Compared with the blank control group, AChE activity and content of norepinephrine （NA）, dopamine （DA）, and 5-hydroxytryptamine （5-HT） were significantly decreased in the model group （P 〈 0.01 ）. In contrast, AChE activity and NA, DA, and 5-HT levels significantly increased in the Kangnaoling and high dosage Schisandra chinensis polysaccharide groups （P 〈 0.01）, while NA levels significantly increased in the low dosage Schisandra chinensis polysaccharide group （P 〈 0.01）. Drug treatment improved learning and memory abilities （P 〈 0.01 or P 〈 0.05）. CONCLUSION： Schisandra chinensis polysaccharide significantly increased levels of central neurotransmitters and improved learning and memory in a mouse model of brain aging. The effects of Schisandra chinensis polysaccharide were equal to that of Kangnaoling pellets.

Analysis of Essential Oil from Stems of Schisandra Chinensis(Turc z.) Baill

The essential oil of Schisandra Chinensis(Turc z.) Baill was extracted and analyzed. The essential oil was obtained in a 1.8%(mL/100 g material) yield. Forty eight compounds were isolated and characterized from the oil. The results show that the content(20.42%) of β -pinene of naturally essential oil is the highest among the components of the oil,which has laid a foundation for the futher development and application of the stems of Schisandra Chinensis(Turc z.) Baill .

Study on HPLC method to determine contents of Schisandrin A and Schisandrin B in Schisandra chinensis extraction

The determination method of Schisandrin A and Schisandrin B in Schisandra chinensis was improved with the high performance liquid chromagraphy （HPLC）. The sample was extracted exceedingly in the critical limit of CO2. The retention time of Schisandrin A and Schisandrin B was reduced, with methano/water （75 ： 25） as mobile phase. The wavelength for detection was 254 nm. The R^2 of standard curve was 0.9998 and the relative standard deviation was 2.31% and 3.17% with the recovery of 96.45% and 97.37%, respectively. The result shows that the rate of veracity of this method is higher and it proves that the determination method of Sehisandrin A and Schisandrin B in Schisandra chinensis is a feasible method.

How Schisandrae Fructus Benefits the Body: Mechanisms in Traditional Chinese Medicine and Modern Medicine

Schisandrae chinensis Fructus (SF) is a commonly used herb in Traditional Chinese Medicine (TCM). According to TCM theory, SF can invigorate Qi in the liver and other visceral organs through the meridian system. Furthermore, the liver’s pivotal role in regulating the functions of various visceral organs helps explain how SF can promote holistic health benefits. The main active ingredient of SF, schisandrin B (Sch B), has been found to improve mitochondrial ATP production and enhance glutathione redox status in multiple organs. This could account for the overall protective effects of Sch B on organs. Due to its stronger impact on liver function, the positive influence of Sch B on different organs may be facilitated by signal molecules originating from the liver.

Pharmacological effects of Schisandra Chinensis Fructus polysaccharide on intestinal protection

Schisandra Chinensis Fructus(SCF)is the fruit of Schisandra chinensis(Turcz.)Baill.,a perennial vine.It was first recorded in Shen Nong′s herbal classic and has a long application history.Studies have shown that SCF has anti-inflammatory,protective liver,antioxidant,antibacterial and other pharmacological effects.Ancient prescriptions are commonly used in the treatment of chronic diarrhea and other intestinal diseases and diabetes.Modern clinical pharmacology features of SCF polysaccharide(SCFP)in diabetes,liver diseases,enteritis and other aspects have achieved excellent results.Gut is an important digestive organ of human body,but intestinal diseases are varied,including Crohn′s disease,ulcerative colitis,intestinal flora imbalance,etc..It is a chronic and non-specific inflammatory disease.The disease is persisted for a long time and the incidence rate is expected to rise.Most of the symptoms are recurrent diarrhea,bloody stool and abdominal pain.It is considered by the World Health Organization as a refractory disease.At present,there is little possibility of complete cure,which is closely related to complex environmental factors,eating habits and heredity.In recent years,clinical studies have found that SCFP has a variety of pharmacological effects on intestinal protection.①Reduce inflammatory factors:intestinal mucositis is a common adverse reaction in patients with chemotherapy.The development of mucositis is related to pro-inflammatory factors such as tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),IL^(-1)β,Interferon-γ(IFN-γ).SCFP can significantly reduce IL-6 TNF-α,IL^(-1)β,and IL-8,as well as the accumulation of T cells in the process of resisting apoptosis,reduce the inflammatory reaction and protect the damage to villi and crypts,improve the symptoms of small intestinal mucositis caused by weight loss and diarrhea.②Promote immunoglobulin A secretion:intestinal mucosal immunity is the first line of defense of the body′s immune system.Its main antibody is secretory immunoglobulin A,which can destroy and phagocytize microorganisms,bacteria and viruses.SCFP can improve intestinal immunity by increasing the number and activity of T lymphocytes,promoting the secretion of secretory immunoglobulin A,and affecting the activity of a variety of cytokines.③Regulation of intestinal flora:the flora in the intestine has the functions of auxiliary nutrient absorption,biological antagonism and immune regulation,and can form a natural barrier for the host's intestine.When the human intestinal flora is disordered,probiotics will be greatly reduced,harmful bacteria will proliferate and destroy the intestinal environment.Under these conditions,the intake of SCFP significantly increased the number of beneficial bacteria such as bifidobacteria and lactobacillus,and significantly decreased the number of conditional pathogens such as enterococcus and escherichia coli,indicating that SCFP can indeed regulate the intestinal disorder caused by lincomycin hydrochloride to a certain extent. This may be because beneficial bacteria in the intestine metabolize polysaccharides produce short chain fatty acids such as lactic acid and acetic acid, which reduces the pH value in the intestine and inhibits the growth of enterococcus and Escherichia coli. In conclusion, SCFP can treat and protect intestinal diseases to a certain extent, which provides a favorable basis for the treatment of intestinal diseases.

Metabolism alternation of schisandra chinensis in carbon tetrachloride-intoxicated rats and patients with advanced hepatocellular carcinoma

OBJECTIVE To study the pharmacokinetics change of schisandra chinensis under the pathological condition of liver dysfunction for safe and rational use of herbal medicines.METHODS The metabolism of four effective lignans from schisandra chinensis(SC),schisandrin,schisantherin A,deoxyshisandrin and γ-schisandrin was studied using microsomes from patients with advanced hepato.cellular carcinoma.In situ intestinal and hepatic perfusions were conducted to clarify the contributions from impairments of gut and liver on the pharmacokinetics of the four schisandra lignans in CCl4-intoxi.cated rats.The metabolism in rat and human liver microsomes and transport in Caco-2 monolayer cell model were studied to reveal the key factors for the in vivo disposition of the four lignans.RESULTS When SC alcoholic extract was orally administrated to CCl4-intoxicated rat for a short term(4 d),the pharmacokinetics of four active SC lignans was significantly changed while its hepatoprotective effect was not obviously observed.The plasma concentrations of the four schisandra lignans were dramatical.ly elevated compared with the control.The Cmax,AUC and MRT were all increased or prolonged signif.icantly while parameter CLz/F was obviously reduced in rat pretreated with CCl4.In hepatic perfusion study and liver microsomes incubation,it was found that the hepatic metabolism of the four lignans was markedly decreased mainly due to the activity reduction of multiple CYP450 isoenzymes involved the metabolism,which,eventually,might lead to the alternation of their pharmacokinetic profiles in CCl4-intoxicated rats or patients with advanced hepatocellular carcinoma.CONCLUSION The pharmacoki.netic studies of SC components in pathological situation of liver dysfunction are expected to provide useful data for rational and safe application of SC preparations in clinic or further pharmacological and toxicological research.

北五味子(Schizandra Chinensis Baillon)的营养学研究

北五味子(Schizandra Chinensis Baillon)分布在东北东部山区,在林下林间大量生长,野生资源丰富。是我国东北地道的中草药,并为医药学家视为佳品。前人对五味子的药理学、成药学作了大量的研究工作,但关于它的营养学研究尚未见报导。作者利用先进的科学仪器定量测定了原汁的各种营养素的含量。结果表明五味子营养素齐全,维生素、微量元素、有机酸含量丰富,是制造天然保健饮料的优秀原料。

Transcriptome-wide analysis of PIP reductase gene family identified a phenylpropene synthase crucial for the biosynthesis of dibenzocyclooctadiene lignans in Schisandra chinensis

Phenylpropenes,such as isoeugenol and eugenol,are produced as defend compounds,floral attractants,and flavor constituents by phenylpropene synthases belonging to the PIP reductase family.Moreover,isoeugenol is proposed to be involved in the biosynthesis of dibenzocyclooctadiene lignans,the main active compounds of Schisandra chinensis(Turcz.)Baill.fruits(SCF).S.chinensis,a woody vine plant,is widely used for its medicinal,horticultural,edible,and economic values.In this study,nine ScPIP genes were identified and characterized from the transcriptome datasets of SCF.The expression profiles revealed that ScPIP genes were differentially expressed during different developmental stages of SCF.Three ScPIPs were selected and cloned as candidate genes encoding phenylpropene synthases according to phylogenetic analysis.ScPIP1 was proved to function as isoeugenol synthase(IGS)and designated as ScIGS1 through in vivo functional characterization in Escherichia coli.Subcellular localization analysis demonstrated that ScIGS1 was localized in both the cytoplasm and nucleus.The three-dimensional(3D)model of ScIGS1 was obtained using homology modeling.Site-directed mutagenesis experiments revealed that the substitution of residues at positions 110 and 113 impacted the product specificity of ScIGS1 and the mutation of Lys157 to Ala abolishing catalytic function.Moreover,the kcat values of mutants were lower than that of ScIGS1 using a deep learning approach.In conclusion,this study provides a basis for further research on PIP reductases and the biosynthetic pathway of dibenzocyclooctadiene lignans.

基于生态位模型的南、北五味子潜在适生区预测及其影响因子研究

基于184条南五味子(Schisandra sphenanthera)地理分布记录、129条北五味子(Schisandra chinensis)地理分布记录以及23个环境因子数据,采用MaxEnt生态位模型预测南、北五味子潜在适宜分布区,评估了制约其地理分布的主要环境因子。结果表明,北五味子和南五味子MaxEnt模型训练集和验证集曲线下面积(AUC)均大于0.90,建立的模型预测结果准确。北五味子最佳生境主要分布在辽宁省和吉林省的长白山区域以及北京市;南五味子的最佳生境主要分布在秦巴山区内,以及湖南省武陵山脉,郴州市、株洲市和衡阳市,浙江省金华市等地区。限制北五味子生长的重要环境因子变量分别是最湿月份降水量、最热月份最高温度、土壤酸碱度、最暖季平均降水量、最冷季平均降水量和昼夜温差月均值;限制南五味子分布的环境因子变量分别是最冷月份最低温温度、年平均降水量、温度变化方差和降水量变化方差。南、北五味子的潜在适生区呈区域化分布。

五味子提取物对仔猪生长性能、肠道形态和免疫功能的影响

文章旨在探究五味子提取物对仔猪生长性能、肠道形态和免疫功能的影响。试验将400头28日龄健康、体况良好的三元杂交猪随机分为4组,每处理10个重复,每重复10头。试验日粮分别向基础日粮中添加0、200、400和800 mg/kg的五味子提取物,预饲期3 d,正试期28 d。结果表明:补充400和800 mg/kg五味子提取物可显著提高仔猪6.25%和4.61%的终末体重(P<0.05);显著提高仔猪15.67%和10.95%的平均日增重(P>0.05);显著降低仔猪10.41%和9.09%的料重比(P<0.05);此外,0~800 mg/kg的五味子提取物对仔猪采食量无显著影响(P>0.05)。800 mg/kg的五味子提取物可显著提高仔猪血清IgM含量(P<0.05);200~800 mg/kg的五味子提取物可显著提高仔猪血清IgG含量(P<0.05);400 mg/kg的五味子提取物可显著降低仔猪血清IL-1β含量(P<0.05);此外,五味子提取物对仔猪血清IgA和TNF-α含量无显著影响(P>0.05)。400和800 mg/kg五味子提取物组仔猪空肠绒毛高度显著提高(P<0.05);400 mg/kg五味子提取物组仔猪空肠隐窝深度及V/C值显著提高(P<0.05);400 mg/kg五味子提取物组仔猪回肠绒毛长度及V/C值显著提高(P<0.05);此外,0~800 mg/kg五味子提取物对仔猪十二指肠肠道形态、回肠隐窝深度均无显著影响(P>0.05)。在本试验条件下,五味子提取物最适添加量为400 mg/kg。

基于化学计量学和指纹图谱的辽宁道地药材北五味子质量评价研究

目的 采用指纹图谱与化学计量学相结合的方法,评价辽宁岫岩产北五味子的质量。方法 采用HPLC法,柱温30℃,流速1 mL/min,流动相采用水-乙腈梯度洗脱,检测波长220 nm,对10批辽宁岫岩五味子基地生产的北五味子建立指纹图谱,运用聚类分析(hierarchical cluster analysis, HCA)、主成分分析(principal component analysis, PCA)及正交偏最小二乘法-判别分析(orthogonal partial least squares-discriminant analysis, OPLS-DA)进行化学模式识别分析。结果 建立了岫岩产北五味子的指纹图谱,相似度为0.970^0.999,共标定了29个共有峰,指认了14个成分;HCA分析10批北五味子可分为2类;PCA共得到6个主要成分,其累计方差贡献率为95.5%;OPLS-DA表明五味子甲素、戈米辛G、五味子丙素、五味子醇乙等11个成分可能是影响北五味子质量的差异性标志物。结论 研究所建立的指纹图谱结合化学模式识别分析,方法准确、稳定、可靠,可用于北五味子药材的质量控制研究。

基于UPLC-Q-TRAP-MS分析养心安神药对酸枣仁-五味子配伍煎煮前后10个指标成分含量变化

目的分析养心安神药对酸枣仁-五味子配伍煎煮前后主要指标成分差异变化,并建立含量测定方法。方法采用Welch Ultimate XB-C_(18)色谱柱(2.1 mm×50 mm,5μm),以0.1%甲酸水溶液(A)-乙腈(B)为流动相,流速0.4 mL/min柱温40℃,梯度洗脱;质谱采用电喷雾离子源(ESI),多反应监测离子扫描模式(MRM)。分别对该药对单煎液、单煎合并液与合煎液中主要指标成分酸枣仁皂苷A、斯皮诺素、五味子醇甲、五味子酯甲、五味子甲素、五味子乙素、芦丁、槲皮素、山柰酚、山柰酚-3-O-云香糖苷进行含量测定,并分析药对配伍前后主要指标成分的含量变化差异。结果以上10种成分在一定浓度范围内均呈现良好的线性关系;精密度、重复性和稳定性RSD均良好;平均加样回收率在95.0%~102.1%,RSD为0.03%~3.7%。本次实验结果与前期试验认为酸枣仁与五味子在合煎后,主要指标成分降低保持一致性。结论多指标成分同时测定为该药对配伍质量全面控制提供新的方法并为其药效物质基础的深入研究提供依据。

基于网络药理学探究北五味子治疗缺血性脑卒中机制

目的:探究北五味子治疗缺血性脑卒中的有效活性物质和具体机制。方法:通过中药系统药理学技术平台筛选出北五味子的有效活性成分,应用Swiss target prediction数据库获得有效活性物质的作用靶点;通过人类基因数据库和在线人类孟德尔遗传数据库获得缺血性脑卒中的作用靶点;通过韦恩图获得共同靶点,借助String数据库进行蛋白质互作网络的建立;通过DAVID数据库进行基因本体(GO)分析获得生物学过程条目、分子功能条目和细胞组成条目;通过基因组百科全书(KEGG)通路富集分析,获得北五味子发挥作用的潜在作用机制;通过Cytoscape 3.8.2软件建立北五味子有效成分-靶点-通路网络。结果:北五味子具有7种治疗缺血性脑卒中的有效成分,18个核心靶点;通过GO分析发现北五味子治疗缺血性脑卒中涉及220个生物学过程条目,27个细胞组成条件,34个分子功能条目;通过KEGG通路富集分析发现北五味子发挥治疗缺血性脑卒中涉及到VEGF信号通路、PI3K-Akt信号通路、脂质与动脉粥样硬化通路和TNF信号通路等;北五味子治疗缺血性脑卒中网络涉及到35个节点、184条边,平均相邻节点数目为10.514,网络中心度为0.309。结论:北五味子有效成分、多靶点、多通路起到防治缺血性脑卒中的作用。本次研究预测的北五味子防治缺血性脑卒中潜在机制可为开展实验研究及临床应用提供依据。

指纹图谱技术结合化学计量学评价五味子醇提物质量探索研究

目的:优化五味子乙醇提取工艺,建立其醇提物HPLC指纹图谱并结合化学计量学综合分析其质量。方法:基于单因素试验,采用L9(34)正交试验设计,系统研究了乙醇浓度、提取时间、提取次数及料液比等因素对五味子中五味子醇甲和五味子乙素的提取率产生的影响。按最优提取工艺制备9批五味子乙醇提取物,建立其高效液相色谱(HPLC)指纹图谱。以共有峰峰面积为变量,结合化学计量学方法包括系统聚类分析(HCA)、主成分分析(PCA)法对9批五味子醇提物质量进行分析。结果:确定最佳提取工艺为:采用80%乙醇作为溶剂,料液比为1:10,进行2次回流提取,每次持续1 h。在9批五味子提取物指纹图谱中共有32个共有峰,并成功鉴定出X18(五味子醇甲)、X29(五味子甲素)和X32(五味子乙素),相似度高于0.94。HCA和PCA结果均显示9批样品有聚为四类的趋势,共有峰X1、X2、X3、X13、X17、X19、X21和X30是引起样品间差异的主要化学成分。结论:所优选的提取工艺稳定可行,不同批次五味子提取物的质量一致性较高,但不同样品间仍存在一定差异。

基于网络药理学探析五味子防治稽留流产的机制及验证

目的:基于网络药理学方法分析五味子防治稽留流产的潜在作用机制,并通过实验进行验证。方法:运用系统药理学方法查找并从中药系统药理学数据库和分析平台(Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform,TCMSP,https://old.tcmsp-e.com/tcmsp.php)筛选五味子的活性成分及靶点,查找稽留流产相关基因,确定五味子防治稽留流产的靶点。利用Cytoscape构建“药物成分-靶点”网络,筛选关键化合物。利用String建立蛋白质-蛋白质相互作用(protein-protein interaction,PPI)网络,并通过Cytoscape-CytoNCA拓扑分析筛选核心靶点。通过对靶基因进行基因本体(gene ontology,GO)和京都基因与基因组百科全书(kyoto encyclopedia of genes and genomes,KEGG)分析,以预测其可能的信号通路及机制。最后借助人绒毛膜外滋养层细胞系(HTR-8/SVneo)利用实时荧光定量聚合酶链反应(real-time fluorescence quantitative polymerase chain reaction,qRT-PCR)检测和蛋白质印迹(Western blotting)检测进行机制验证。结果:从TCMSP数据库中鉴定了7种五味子活性成分。PPI网络表明雄激素受体(androgen receptor,AR)、雌激素受体(estrogen receptor,ER)、环氧合酶-2(cyclooxygenase-2,COX-2)、核受体辅激活因子2(nuclear receptor coactivator 2,NCOA2)和乙酰胆碱酯酶(acetylcholine esterase,ACHE)可能是五味子防治稽留流产的核心作用靶点。GO富集分析获得44个细胞生物学过程,KEGG途径富集分析获得2个相关信号通路,主要包括甲状腺激素信号通路和雌激素信号通路;在利用细胞模型验证机制过程中发现,二羟环氧苯并芘[benzo(a)pyren-7,8-dihydrodiol-9,10-epoxide,BPDE]染毒后的细胞中AR、ER和COX-2的蛋白质和(或)mRNA水平高于正常细胞,而先使用五味子乙素(Schisandrin B,Sch B)预处理后再染毒BPDE的细胞中AR、ER和COX-2的蛋白质和(或)mRNA水平均较未用Sch B预处理的细胞降低,而NCOA2和ACHE的蛋白质和(或)mRNA水平变化不明显。结论:五味子可通过抗炎和调节AR、ER发挥防治稽留流产的作用,这一保护作用可能是通过雌激素信号通路来实现的。