Pharmacodynamic evaluation of Cordyceps sinensis (Berk.) Sacc. for ischemic stroke in rats and potential mechanism through network pharmacology and molecular docking

Objective:To explore the therapeutic potential of Cordyceps sinensis(Berk.)Sacc.(C.sinensis,Dong Chong Xia Cao)in an ischemic stroke(IS)model and predict its possible mechanism through network pharmacology.Methods:Thirty-three SpragueeDawley rats were randomly divided into the Sham,model,and C.sinensis groups.After 5 days of pre-treatment,the model group and the C.sinensis group were sub-jected to middle cerebral artery occlusion(MCAO)modeling.Effect of C.sinensis on MCAO rats was evaluated by comparing cerebral infarct size,neurological function,cerebral water content,pathological changes,and certain biochemical indicators.Intersection targets between C.sinensis and IS was screened using network pharmacology analysis.Relationship among core components,targets and pathways of C.sinensis in treating IS was constructed through network pharmacology analysis and further verified by molecular docking.Finally,the DAVID v8.8 database was used for performing GO analysis and KEGG pathway enrichment analysis by importing the intersection targets.Results:Compared with the model group,C.sinensis significantly reduced the volume of cerebral infarction(P=0.026),the cerebral water content(P¼.0013),the mNSS score(P<0.001),and the levels of IL-17(P=0.031),TNF-α(P=0.016),MDA(P=0.014),and glutamate(P=0.014)in serum,while upregulating the level of SOD in serum and improving the pathological morphology in MCAO rat ischemic brains.The results of network pharmacology analysis showed that core targets(such as CASP3,PTGS2,and PPARG)and the main enrichment pathways(IL-17,AGE-RAGE,and TNF signaling pathways)were regulated by 30 chemical components of C.sinensis,which effectively treated IS in MCAO rats.Conclusion:The results of this study showed that C.sinensis effectively interfered with MCAO rats,and the mechanism may be related to the regulation of blood lipids and to anti-apoptosis and anti-inflammatory effects.

Mechanism of Cordyceps sinensis in atherosclerosis treatment based on network pharmacology and molecular docking analysis

Background:The present study intented to delve into the molecular mechanism of Cordyceps sinensis(C.sinensis)in treating atherosclerosis by combining network pharmacology and molecular docking analysis.Methods:We searched the databases including Traditional Chinese Medicines Systems Pharmacology Database and Analysis Platform,PubChem,and PharmMapper to screen out the active chemical ingredients of C.sinensis and the corresponding targets.The String database was used for the matching normalization of results,and the software Cytoscape 3.7.2 was adopted to establish the C.sinensis-active components-targets of action-disease network.The databases of Online Mendelian Inheritance in Man database,GeneCards,Therapeutic Target Database,and DisGNET were searched to yield the major targets of atherosclerosis(AS),which were matched with the active component targets of C.sinensis to identify the potential therapeutic targets.The String database was utilized to set up the protein-protein interaction network,and Cytoscape software was applied for topological analysis,which was followed by the Gene Ontology analysis and Kyoto Encyclopedia of Genes and Genomes signaling pathway analysis based on the DAVID database.Finally,the core components of C.sinensis and the targets of action were confirmed via molecular docking on AutoDock Vina and PyMOL.Results:In total,7 bioactive ingredients of C.sinensis were identified from Traditional Chinese Medicines Systems Pharmacology Database and Analysis Platform database and 319 predicted targets were obtained,231 of which were associated with AS.The core targets involved in AS treatment with C.sinensis included MAPK1,SRC,PIK3R1,AKT1,and HSP90AA1.The enrichment analysis unveiled the primary pathways involved in these processes,such as pathways in cancer and lipid and atherosclerosis.Moreover,through molecular docking,it was found that the active ingredients of C.sinensis presented with strong binding capacities with their corresponding targets,and the strongest binding capacity was observed between peroxyergosterol and SRC.Conclusions:The present study revealed at the molecular level that C.sinensis played its role in AS treatment through multiple drug active components,targets of action and pathways,which would point out the direction and provide theoretic basis for future research.

Cultivated Cordyceps: A Tale of Two Treasured Mushrooms

Ophiocordyceps sinensis and Cordyceps militaris both contain many bioactive compounds that confer potential therapeutic benefits. This review discusses the possible use of cultivated C. militaris as an effective substitute for native O. sinensis in the face of ever-increasing prices of O. sinensis because of its short supply. On the one hand, cultivated C. militaris contains higher levels of cordycepin when compared with that of wild-type O. sinensis and cultivation of C. militaris has been shown to be capable of reducing the risk of heavy metal contamination. On the other hand, there is a paucity of robust in vivo studies and randomized controlled tests comparing the pharmacology and use of C. militaris and O. sinensis. For extraction of cordycepin as western-style tablets, the use of cultivated C. militaris rather than O. sinensis represents the most appropriate future approach. For many other purposes, comparative pharmacology and clinical trials are in urgent needs.

Determination of Cordycepin in Cordyceps kyushuensis by Capillary Electrophoresis and its Antitumour Activity

A simple, rapid and low-cost method of determination for cordycepin in Cordyceps kyushuensis by capillary zone electrophoresis (CZE) was developed. Based on the finding that there is a high concentration of cordycepin in both natural and cultured Cordyceps kyushuensis, the in vitro antitumor activity of cordycepin and the water extracts of Cordyceps kyushuensis has been investigated. This is the first report about the antitumor effect of Cordyceps kyushuensis.

<i>Cordyceps sinensis</i>Acts As an Adenosine A₃Receptor Agonist on Mouse Melanoma and Lung Carcinoma Cells, and Human Fibrosarcoma and Colon Carcinoma Cells

Cordyceps sinensis, a parasitic fungus on the larva of Lapidoptera, has been used as a traditional Chinese medicine. We previously reported that the growth of B16-BL6 mouse melanoma (B16-BL6) cells and mouse Lewis lung carcinoma (LLC) cells was inhibited by cordycepin (3’-deoxyadenosine), an ingredient of Cordyceps sinensis, and its effect was antagonized by MRS1191, a selective adenosine A3 receptor (A3-R) antagonist although adenosine (up to 100 μM) had no effect on the growth of B16-BL6 and LLC cells. In this study, we investigated whether water extracts of Cordyceps sinensis (WECS) inhibit the growth of B16-BL6 cells, LLC cells, HT1080 human fibrosarcoma (HT1080) cells and CW-2 human colon carcinoma (CW-2) cells via their A3-R. As a result, the growth of all cell lines were potently inhibited by WECS (10 μg/mL) and the inhibitory effect of WECS was significantly antagonized by MRS1191 (1 μM). Furthermore, WECS included 2.34% w/w cordycepin and 0.12% w/w adenosine as components according to the HPLC- ECD system. In conclusion, WECS inhibited the proliferation of four cancer cell lines by stimulation of A3-R and the main component in WECS with anticancer action might be cordycepin instead of adenosine.

The Effect of the Hot Water Extracts of the <i>Paecilomyces hepiali </i>and <i>Cordyceps militaris</i>Mycelia on the Growth of Gastrointestinal Bacteria

The gastrointestinal health is very important aspect concerning human health. It refers to nutrient and vitamin absorption, food digestion and various infectious diseases. The Paecilomyces hepiali and Cordyceps militaris are highly enriched with cordycepin and ergosterol which are considered as anti bacterial substances. Present study finds out comparative effect of hot water extract of particular fungal material on growth of six species of gastrointestinal bacteria that belong to both aerobic and anaerobic and, consist with harmful and commensal categories. The appropriate concentration level of hot water extract of both strains was identified. The individual specific bacterial growing media were prepared and calculated;amounts of bacteria cultures were inoculated by using micro pipettes. The optical density and number of bacterial colonies were measured after 24 hours. The pure mycelial extract of P. hepiali with 2 × 10-3 g/ml of concentration has significant effect on depleting the growth of E. coli, E. faecalis, S. aureus, L. gasseri and B. ovatus bacteria. B. longum has no significant effect by particular extract. Same type of extract of C. militaris has significantly reduced the growth of every bacteria used in this study. Hot water extract of C. militaris cultivated on soy bean has significant growth retardation toward E. coli, E. faecalis, S. aureus and L. gasseri. It has stimulated the growth of B. ovatus and B. longum which are considered as beneficial bacteria for human gut. This study shows that extracts of both mycelia include antimicrobial substances like cordycepin and ergosterol which can be used as food supplements to enhance human gut health.

Effects of heat on the biological activity of wild Cordyceps sinensis

Background:Current methods of extending the storage time of wild Cordyceps sinensis adversely affect the nutritive and medicinal value of the product.Thus,this study was designed to investigate the effects of heat treatment,a relatively safe storage extension method,on the biological activity of wild C.sinensis.Methods:Samples were heated to 60,80,or 100℃ for 15,30,or 60 minutes.SOD activity in wild C.sinensis before and after heating was assayed using a standard colorimetric assay.Deoxyribonuclease (DNase) activity was measured using the plasmid-nicking assay.Cordycepin content was analyzed using HPLC.Polysaccharide content was measured using the phenol sulfuric method.The Student's t-test was used for comparison.Results:After heating at 60,80,100℃ for 15,30,60 minutes,respectively,no significant reduction in DNase activity or polysaccharide dissolution was noted (P >.05).Interestingly,heating at 80℃ for 30 minutes led to a significant increase in the SOD activity of C.sinensis (P <.05).In addition,heating at 60℃ for 60 minutes or at 80℃ for 15 minutes significantly increased cordycepin dissolution (P <.05).Other heat treatments had no significant effects on SOD activity or cordycepin dissolution (P >.05).Conclusions:These results suggested that heat treatment does not adversely affect SOD or DNase activity,polysaccharide content,or cordycepin dissolution.Thus,heat treatment might be a safe processing method to extend the storage time of wild C.sinensis without compromising biological activity.

冬虫夏草(Cordyceps sinensis)的随机扩增多态DNA及其遗传分化

本文对来自青藏高原3个区域5个具有代表性地方的13个冬虫夏草(Cordyceps sinensis[Berk.] Sacc.)样本进行随机扩增多态DNA(RAPD)分析。19个随机引物获得的RAPD谱带清晰并呈现多态,单个引物获得的RAPD片段数在3～10个之间。该19个引物在每个样本中扩增的RAPD片段总数平均约为65个。基于遗传距离分析,受试的13个冬虫夏草样本中,来自同一地方的样本间遗传差异甚微,同一区域不同地方的样本间遗传差异较大,不同区域的样本间遗传差异最大。这说明冬虫夏草地理群体间存在着遗传分化。应用UPGMA和NJ方法构建的分子系统树显示,来自5个地方的冬虫夏草实际上可以归并为显著不同的3个组,对应于样本来源的3个区域,提示RAPD标记在冬虫夏草群体中有显著的地区特异性。我们的结果还表明,冬虫夏草地理群体间的遗传差异度与地理距离呈正相关。因此,RAPD作为有效的遗传标记,可用于研究冬虫夏草的遗传多样性、起源以及系统演化等。

冬虫夏草(Cordyceps sinensis)菌丝体固体发酵粉化学成分的分析

本研究对一种冬虫夏草(Cordycepssinensis)菌丝体固体发酵粉的化学成份进行了分析,包括粗成份,生物活性成份,脂肪酸,氨基酸,金属元素等。冬虫夏草菌丝体固体发酵粉中3′-脱氧腺苷含量40.81μg/g、腺苷含量745.4μg/g;甘露醇的含量为165.8mg/g,多糖含量为157.3mg/g;谷氨酸,天门冬氨酸,苯丙氨酸是三大类主要氨基酸;不饱和脂肪酸是优势脂肪酸,以亚油酸,油酸为主。和文献记录的结果相比,虫草发酵粉中3′-脱氧腺苷,甘露醇的含量明显高于冬虫夏草子实体中相应成份的含量;各种氨基酸的含量和水平与冬虫夏草子实体中相应氨基酸的含量和水平相当;虫草发酵粉中的其它化学成份和冬虫夏草子实体中相应成份基本一致。

Cordycepssinensis CAMT63341液态发酵多糖对小鼠若干行为学指标及免疫器官指数的影响

为评价虫草菌(Cordyceps sinensis)CAMT 63341液态发酵多糖对小鼠生长、情绪、认知和免疫的影响,将60只健康雄性昆明小鼠随机分成空白对照、灵芝多糖对照和Cordyceps sinensis CAMT 63341发酵多糖高、中、低剂量饲喂组连续喂养30 d,然后行高架十字迷宫、穿梭箱、迟发型超敏反应实验,并记录小鼠体质量和免疫器官指数。结果表明:与空白对照组相比,Cordyceps sinensis CAMT 63341发酵多糖可剂量依赖性促进小鼠生长,增加高架迷宫开放臂次数和延长停留时间,增强穿梭箱实验中主动回避能力和缩短被动回避潜伏期,提升胸腺和脾脏器官指数,其中,高剂量组达到显著(P<0.05)或极显著水平(P<0.01),接近灵芝多糖对照组,提示Cordyceps sinensis CAMT 63341液态发酵多糖具有一定的促进生长、改善情绪、增强认知和提升免疫功能。

北冬虫夏草(Cordyceps militaris)的研究应用概况(综述)

主要综述了近二十年来国内外对北冬虫夏草的研究应用概况，详细地介绍了北冬虫夏草及其代谢物质“虫草素”的来源，及在功能、药化、药理、临床应用方面的研究进展，北冬虫夏草的生物学特性和工厂化生产研究情况，以及影响北冬虫夏草人工高产栽培与工厂化生产的关键问题，同时也展示了北冬虫夏草的开发利用前景。

蛹虫草Cordyceps militaris JN168产虫草素液态发酵条件的优化

利用单因素筛选和响应面法对蛹虫草Cordyceps militaris JN168产虫草素的液态发酵培养基进行优化,以确定蛹虫草产虫草素的最佳发酵培养基配方。结果表明,蛹虫草产虫草素的最佳碳源为葡萄糖,最适质量浓度为40 g/L;最佳氮源为牛肉膏,最适质量浓度15 g/L;加入的无机盐及其添加量分别为MgSO40.76 g/L,K2HPO40.63 g/L,CaCl20.66 g/L,Na2HPO40.67 g/L。优化后发酵液中虫草素质量浓度达到633.47 mg/L,是优化前的6倍。

蛹虫草(Cordyceps militaris)对药物引起免疫抑制的拮抗作用

实验研究蛹虫草对环磷酰胺造成的小鼠免疫抑制的作用。实验选用天然冬虫夏草为阳性对照。给药方式为每天灌胃 0 .2 g生药水煎剂 ,7d后测定其生物学活性。实验结果表明 :小鼠腹腔巨噬细胞的吞噬率由对照组的 13.97%提高到 2 2 .16 %;骨髓细胞的微核数目由对照组的 16 .6×10 -3 降低到 12 .3× 10 -3 ;外周血白细胞由对照组的 70 %提高到 83%。结果证实 :蛹虫草具有明显的拮抗由环磷酰胺造成的免疫抑制的作用 ,与冬虫夏草比较有相似的生物学活性 ,某些作用明显高于冬虫夏草 ,为蛹虫草的医学应用提供了一定的理论依据。

蛹虫草Cordyceps militaris JN168菌株的诱变育种及液态发酵产虫草素

利用离子束、亚硝基胍及离子束-亚硝基胍诱变法处理蛹虫草Cordyceps militaris JN168,初步获得了几株较高产虫草素的菌株。通过进一步筛选得到复合诱变菌2,并对该菌的液态发酵培养基组分进行了优化。实验结果表明:蛹虫草液态发酵产虫草素的最佳培养基组分为(g/L),葡萄糖40,酵母浸粉25,Mg SO4·7H2O 0.6,K2HPO4·3H2O 0.6,KH2PO40.6。优化后虫草素产量提高了5倍,最高达1 045.65 mg/L。

北冬虫夏草(Cordyceps militaris)单孢菌株配对后对子实体形成的影响与无性型产孢结构关系

由野生子实体分离的低代菌株“ Ｃ．ｍ．Ｈ９５－１”和“ Ｃ．ｍ．Ｈ９７－Ｇ６”的子囊孢子单孢，配对后产生了较大变异，３１％和５０％的配对菌株具有与亲本菌株相同的遗传特性，而且子实体形成能力及生物产量明显增加。配对菌株“ Ｃ．ｍ．Ｈ９７－２６９３”和“ Ｃ．ｍ．Ｈ９７－２６２７”的子囊孢子再配对后，几乎所有的再配对菌株不能回复到亲本菌株的生长型，其子实体的形成能力及生物产量也低于亲本菌株。实验的结果还表明，在ＰＤＡ组合培养基１上，单孢及配对菌株的无性型产孢结构，对子实体形成有着一定的相关性。

蛹虫草(Cordyceps militaris)菌丝体液体培养基的优化和发酵条件的研究

采用二次回归通用旋转组合设计方法，对蛹虫草菌丝体液体发酵培养基的最佳组合进行了研究，建立了培养基的反应模型，并用实验数据验证了数学模型，对培养基最佳优化组合的各种因子、影响产量的反应规律及交互作用进行了探讨，并在此基础上进行了蛹虫草菌丝体液体发酵条件的研究。结果表明，蛹虫草菌丝体液体发酵的最适条件为：接种量６％（ｖ／ｖ），发酵初始ｐＨ７．０，培养基装量１００ｍｌ／５００ｍｌ三角瓶，发酵温度２４℃～２８℃，发酵周期９６ｈ。

Cu^(2+)、Zn^(2+)胁迫对蛹虫草(Cordyceps militaris)抗氧化酶活性及脂氧化水平的影响

研究了Cu2+、Zn2+对蛹虫草菌丝体抗氧化酶活性及脂氧化水平的影响。结果表明:高浓度的Cu2+抑制菌丝体的生长和细胞内抗氧化酶的活性;高浓度的Zn2+对菌丝体的生长也有抑制作用但抗氧化酶的活性变化不大。脂氧化水平随细胞内过氧化状态的变化而变化。蛹虫草菌丝体对Cu2+、Zn2+有较高的耐受度。

粉被虫草(Cordyceps pruinosa Petch)无性型对苏云金杆菌(Bacillus thuringiensis subsp.galleria Heimpel)抗紫外辐射的保护效应

本文首次报道了粉被虫草的无性型粉被马利娅霉(Mariannaea PruinosaLiang)对苏云金杆菌抗紫外辐射的保护作用,建立了粉被马利娅霉抗紫外辐射物质的苏云金杆菌检测法。用粉被马利娅霉的培养液、菌丝体浸提液及孢梗束浸提液所培养的苏云金杆菌经紫外辐射后,活菌数依次比对照提高26.3％、138.6％、187.8％。揭示了抗辐射物质的含量是孢梗束高,菌丝体次之,培养液最低的趋势。不同来源的粉被马利娅霉菌株抗紫外辐射物质的含量是有差异的。

北冬虫夏草(Cordyceps militaris)室内栽培技术研究

2003年从北京北冬虫夏草(Cordyceps militaris)子实体的子座部分经单孢分离出Ⅱ号菌株.2004年从北京、上海北冬虫夏草新鲜子实体中采用组织分离法分离出Ⅰ号、Ⅲ号和Ⅳ号菌株,同时收集到开封试管菌株Ⅴ号.Ⅱ号菌株在不同培养基(5,6,7号)进行瓶栽试验,并与4个菌株(Ⅰ号、Ⅲ号、Ⅳ号、Ⅴ号)在同一培养基进行瓶栽比较试验.经过50d的观察表明,Ⅱ号菌株在5号和6号培养基上所产生子实体及每瓶平均数量,平均长度,平均鲜重和干重都比在7号培养基上良好;Ⅰ号、Ⅲ号、Ⅳ号菌株在5号培养基上生的子实体无论每瓶平均数量、平均长度、平均鲜重和干重都明显比Ⅴ号菌株子实体情况优良.

中国虫草Cordyceps sinensis的研究Ⅲ多样性初探

就中国虫草分类定种中的个种多样化趋势，结合产区生态和物种演化提出新的认识，即中国虫草是青藏高原及其相邻余脉特殊区域内物种适应与分化的结果，同时其自身也呈进一步分化的趋势．