Prediction of potential targets and molecular mechanisms of Tripterygium hypoglaucum for oral lichen planus based on network pharmacology and molecular docking

Objective:To explore the potential targets and molecular mechanisms of tripterygium hypoglaucum for oral lichen planus(OLP)by applying network pharmacology and molecular docking technology.Methods:The active ingredients and targets of tripterygium hypoglaucum were screened.OLP-related targets were predicted.The Protein-Protein Interaction(PPI)network was constructed for the intersection targets of tripterygium hypoglaucum and OLP.The“OLP-target-molecule-Tripterygium hypoglaucum”network was constructed and visualized.The intersection genes were screened for gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses.Molecular docking analysis and visualization were performed.Results:15 active ingredients and 78 targets of tripterygium hypoglaucum were obtained.9109 OLP-related targets were screened,and 54 intersection genes of tripterygium hypoglaucum with OLP were obtained.The top 10 key targets were screened from the constructed PPI network.The top 10 active ingredients of tripterygium hypoglaucum were screened from the constructed“OLP-targets of actionactive ingredients-tripterygium hypoglaucum”network.The GO and KEGG analyses of the 54 intersection targets indicated that tripterygium hypoglaucum may play a therapeutic role by regulating one carbon pool by folate,pathways in cancer,et al.Molecular docking analysis showed that dihydrofolate reductase(DHFR),phosphoribosylglycinamide formyltransferase(GART),estrogen receptor 1(ESR1),et al are the key targets for the treatment of OLP in tripterygium hypoglaucum.Conclusion:The potential key targets and molecular mechanisms of tripterygium hypoglaucum in treating OLP provide a theoretical basis for new drug development and clinical applications.

In vitro Anti-viral Activity of the Total Alkaloids from Tripterygium hypoglaucum against Herpes Simplex Virus Type 1

Herpes simplex virus type 1 (HSV-1) is a commonly occurring human pathogen worldwide. There is an urgent need to discover and develop new alternative agents for the management of HSV-1 infection. Tripterygium hypoglaucum (level) Hutch (Celastraceae) is a traditional Chinese medicine plant with many pharmacological activities such as anti-inflammation, anti-tumor and antifertility. The usual medicinal part is the roots which contain about a 1% yield of alkaloids. A crude total alkaloids extract was prepared from the roots of T. hypoglaucum amd its antiviral activity against HSV-1 in Vero cells was evaluated by cytopathic effect (CPE) assay, plaque reduction assay and by RT-PCR analysis. The alkaloids extract presented low cytotoxicity (CC50 = 46.6 μg/mL) and potent CPE inhibition activity, the 50% inhibitory concentration (IC50) was 6.5 μg/mL, noticeably lower than that of Acyclovir (15.4 μg /mL). Plaque formation was significantly reduced by the alkaloids extract at concentrations of 6.25 μg/mL to 12.5 μg/mL, the plaque reduction ratio reached 55% to 75% which was 35% higher than that of Acyclovir at the same concentration. RT-PCR analysis showed that, the transcription of two important delayed early genes UL30 and UL39, and a late gene US6 of HSV-1 genome all were suppressed by the alkaloids extract, the expression inhibiting efficacy compared to the control was 74.6% (UL30), 70.9% (UL39) and 62.6% (US6) respectively at the working concentration of 12.5μg/mL. The above results suggest a potent anti-HSV-1 activity of the alkaloids extract in vitro.

In vitro porcine brain tubulin assembly inhibition by water extract from a Chinese medicinal herb,Tripterygium hypoglaucum Hutch

瞄准：在汇编和导管素的拆卸过程和它行动的可能的模式上调查 Tripterygium hypoglaucum 兔笼(THH ) 的效果。方法：在试管内猪的大脑导管素集会试金被采用在不同集中分析 THH 的禁止的效果(0.05 microg/L， 0.07 microg/L， 0.09 microg/L ) 。用番红花作原料而制成的植物盐基(0.0025 mmol/L， 0.0050 mmol/L， 0.0075 mmol/L ) 被用作积极控制。结果：THH 能显著地禁止根本测试的孤立的猪的大脑导管素的集会集中。结论：THH 能够经由导管素聚合抑制小径在哺乳动物导致非整倍性并且可以提出基因风险到人。

Comparative Observation on the Effects of Radix Tripterygium Hypoglaucum Tablet and Tripterygium Glycosides Tablet in Treating Erosive Oral Lichen Planus

Objective: To compare the therapeutic effects of Radix Tripterygium hypoglaucum tablet (THT) and Tripterygium glycosides tablet (TGT) in treating erosive oral lichen planus(EOLP).Methods: The patients were randomized into two groups, and they were treated with THT (n=47) or TGT (n=47), respectively. The therapeutic effects were evaluated after 3 months treatment.Results: For the patients of grade 1, the total efficacy in TGT group was 85.71%, compared with 52.38% in THT group, the efficacy was statistically greater in the group receiving TGT (P=0.043). However, for the patients of grade 2, the difference was not statistically significant (P=0.173).Conclusion: TGT is more effective in treating EOLP than THT for grade 1 patients. However, TGT is not suitable for patients of child bearing age.

Research review on the main chemical constituents and pharmacological effects of Tripterygium hypoglaucum (Levl.) Hutch.

Tripterygium hypoglaucum(Levl.)Hutch.(THH),which belongs to Tripterygium genus of Celastraceae family,is widely used as the common Chinese herbal medicine.It has been reported that the chemical constituents of T.hypoglaucum include monoterpenoids,sesquiterpenoids,diterpenoids,triterpenoids,sesquiterpene alkaloids,flavonoids,anthraquinone derivatives,et al.These compounds showed a wide range of bioactivities,such as anti-inflammatory,anti-cancer,immunosuppressive,anti-HIV,anti-fertility and insecticidal activities.In this paper,we summarized the chemical constituents and pharmacological activities of T.hypoglaucum to provide a reference for its further research and developement.

基于网络药理学和分子对接技术预测昆明山海棠治疗口腔扁平苔藓的潜在靶点和分子机制

目的:联合采用网络药理学和分子对接技术,探究昆明山海棠治疗口腔扁平苔藓(oral lichen planus,OLP)的潜在靶点和分子机制。方法:筛选昆明山海棠有效成分及靶点,预测OLP相关靶点,构建昆明山海棠和OLP交集靶点的蛋白质相互作用(protein-protein interaction,PPI)网络,建立“OLP-靶点-分子-昆明山海棠”网络并进行可视化分析,筛选出的交集基因进行基因本体论(gene ontology,GO)功能分析和京都基因与基因组百科全书(kyoto encyclopedia of genes and genomes,KEGG)通路富集分析,分子对接分析及可视化。结果:获得昆明山海棠有效成分15个,靶点78个,筛选得到与OLP相关靶点9109个,昆明山海棠与OLP交集基因54个,从构建的PPI网络筛选获得前10个核心靶点,从构建的“OLP-作用靶点-有效成分-昆明山海棠”网络筛选获得昆明山海棠前10个有效成分,对54个交集靶点的GO分析和KEGG分析结果表明,昆明山海棠可能通过调节一碳叶酸循环、肿瘤信号通路等途径发挥治疗作用,分子对接分析结果表明,二氢叶酸还原酶(dihydrofolate reductase,DHFR)、磷酸核糖甘氨酰胺转甲酰酶(phosphoribosylglycinamide formyltransferase,GART)、雌激素受体1(estrogen receptor 1,ESR1)等是昆明山海棠治疗OLP的关键靶点。结论:本研究获得昆明山海棠治疗OLP的潜在靶点及分子机制,为促进靶向药物的研发及临床应用提供理论基础。

基于网络药理学与分子对接探讨火把花根干预IgA肾病的作用机制研究

运用网络药理学及分子对接技术探讨火把花根治疗IgA肾病(IgA nephropathy,IgAN)潜在分子机制。通过文献检索搜集中药火把花根的化合物成分信息,利用PubChem、SwissADME数据库筛选药物成分,通过Pharmmapper数据库预测药物作用靶点信息。利用DisGenet、DrugBank、GeneCards、OMIM数据库预测IgAN疾病靶点信息。利用R4.2.0 ggvenn包对预测的药物和疾病靶点交互映射、获得交集靶点;利用STRING11.5、Cytoscape 3.9.0对交集靶点进行蛋白互作分析;利用R4.2.0 clusterProfiler包对交集靶点进行GO、KEGG生物信息富集分析,获得火把花根干预IgAN的潜在通路;利用Cytoscape 3.9.0构建药物成分-靶点网络图;使用AutoDock Vina1.1.2对核心成分、受体蛋白结合模式进行分子对接验证。共筛选获得火把花根活性成分19个,预测到火把花根干预IgAN潜在靶点54个,利用cytohubba获得10个核心靶点;GO富集显示潜在靶点主要参与伤口愈合(wound healing)、激素代谢过程(hormone metabolic process)、白细胞黏附(leukocyte cell-cell adhesion)等生物进程;KEGG富集主要集中在癌症中的蛋白聚糖信号通路(Proteoglycans in cancer pathway)、脂质和动脉粥样硬化信号通路(Lipid and atherosclerosis pathway)、FoxO信号通路(FoxO signaling pathway)、内分泌抵抗信号通路(Endocrine resistance pathway)等通路上;分子对接显示Affinity值均<16.7 kJ/mol。通过网络药理学分析,初步预测火把花根潜在活性成分为“雷公藤内酯甲”、“雷公藤内酯乙”、“雷公藤春碱”和“雷公藤红素”等;通过CDK2、AKR1B1、F2和MAPK14等靶点,作用于癌症中的蛋白聚糖、脂质和动脉粥样硬化、FoxO信号通路等信号通路干预IgAN;其中雷公藤内酯甲与AKR1B1结合活性最强;本研究可为进一步探讨火把花根干预IgAN的作用机制提供新思路。

Tripterygium hypoglaucum extract ameliorates adjuvant-induced arthritis in mice through the gut microbiota

Traditionally,Tripterygium hypoglaucum(Levl.)Hutch(THH)are widely used in Chinese folk to treat rheumatoid arthritis(RA).This study aimed to investigate whether the anti-RA effect of THH is related with the gut microbiota.The main components of prepared THH extract were identified by HPLC-MS.C57BL/6 mice with adjuvant-induced arthritis(AIA)were treated with THH extract by gavage for one month.THH extract significantly alleviated swollen ankle,joint cavity exudation,and articular cartilage destruction in AIA mice.The mRNA and protein levels of inflammatory mediators in muscles and plasma indicated that THH extract attenuated inflammatory responses in the joint by blocking TLR4/MyD88/MAPK signaling pathways.THH extract remarkably restored the dysbiosis of the gut microbiota in AIA mice,featuring the increases of Bifidobacterium,Akkermansia,and Lactobacillus and the decreases of Butyricimonas,Parabacteroides,and Anaeroplasma.Furthermore,the altered bacteria were closely correlated with physiological indices and drove metabolic changes of the intestinal microbiota.In addition,antibiotic-induced pseudo germ-free mice were employed to verify the role of the intestinal flora.Strikingly,THH treatment failed to ameliorate the arthritis symptoms and signaling pathways in pseudo germ-free mice,which validates the indispensable role of the intestinal flora.For the first time,we demonstrated that THH extract protects joint inflammation by manipulating the intestinal flora and regulating the TLR4/MyD88/MAPK signaling pathway.Therefore,THH extract may serve as a microbial modulator to recover RA in clincial practice.verRAinclincial practice.

昆明山海棠在微核实验中非整倍体毒性的研究

本文报道以小鼠着丝粒次要卫星ＤＮＡ探针ＦＩＳＨ和抗着丝粒ＣＲＥＳＴ染色，研究了可疑的非整倍体毒剂昆明山海棠（ＴＨＨ）诱导的小鼠ＮＩＨ３Ｔ３细胞微核（ＭＮ）的着丝粒组成情况，结果ＴＨＨ（１０～６０μｇ／ｍｌ）诱导的６２１～６８４％的ＭＮ为ＦＩＳＨ阳性，３５９～４２２％的ＭＮ为ＣＲＥＳＴ阳性，较精确的ＦＩＳＨ结果显示ＴＨＨ具有较强的非整倍体诱发效应，同时认为次要卫星ＤＮＡ探针比ＣＲＥＳＴ染色更适合于ＭＮ的着丝粒检测。

昆明山海棠提取物对血管平滑肌细胞增殖及凋亡的影响

目的探讨昆明山海棠提取物 (THW 4 )对离体血管平滑肌细胞 (VSMC)增殖及凋亡的影响。方法培养家兔主动脉平滑肌细胞 ,加入不同浓度的THW 4进行处理 ,用MTT法测定细胞生长曲线 ,以AnnexinV FITC/PI双标记、透射电镜、原位末端标记法检测细胞凋亡。结果THW 4可以明显抑制VSMC增殖 ,呈剂量和时间依赖性 ,作用 4 8h的半有效抑制浓度 (IC50 )为 15 .6 μg/L ;VSMC在含 10～ 10 0 μg/LTHW 4的培养液中孵育 5 6h ,可检测到细胞早期凋亡 ,且凋亡率随THW 4浓度增加而升高 ;延长孵育时间至 72h ,透射电镜下可观察到典型的凋亡细胞形态学改变 ,原位末端标记法显示DNA片段化增多 ,有核碎裂、固缩等改变 ;细胞周期时相分布显示THW 4主要阻滞VSMC于G2 /M期。结论THW 4能有效抑制体外培养的VSMC增殖及诱导其凋亡 ,提示THW

昆明山海棠对人单核细胞TNFα、IL-8及基因表达的影响

应用体外细胞培养方法、ELISA及RT-PCR技术,观察昆明山海棠对健康人外周血单核细胞IL-8、TNFα释放及IL-8mRNA表达的影响。结果表明昆明山海棠(0.2mg/mL～1mg/mL)可抑制脂多糖(LPS)诱导的外周血单核细胞IL-8、TNFα释放及IL-8mRNA表达,提示昆明山海棠可从转录水平抑制细胞因子表达,从而实现其抗炎机制。

雷公藤与昆明山海棠采样调查报告

目的:通过采样调查,获取雷公藤与昆明山海棠野生资源现状、种植、商品流通等方面的资料。方法:在全国13个样地采集样品,比较雷公藤和昆明山海棠的形态特征与生长习性的差异;调查雷公藤与昆明山海棠的野生资源现状、种植信息、民间混淆种及不同市售药材的商品特征。结果:(1)雷公藤和昆明山海棠主要采集于林下或林地边缘,光照和水分对雷公藤属植物的分布有一定的影响;(2)雷公藤与昆明山海棠的野生资源量正在萎缩,两种植物种植历史较短,种植技术水平较低;(3)雷公藤的采收加工尚缺乏规范,市售药材将雷公藤属植物的根、根茎、茎均作商品药材销售,饮片规格多样。结论:雷公藤与昆明山海棠的野生资源正在萎缩,人工种植、采收加工及市售药材的管理等方面的研究尚需规范深入的开展。

昆明山海棠总生物碱体外抗肿瘤作用的研究

目的:探讨昆明山海棠(THH)总生物碱对10种不同肿瘤细胞系的抑制作用,并筛选出最敏感的肿瘤细胞系。方法:采用XTT法和SRB法对THH总生物碱体外抗肿瘤作用进行初步筛选。结果:THH总生物碱对10种不同肿瘤细胞系生长均有明显的抑制作用,而且对不同肿瘤细胞系抑制作用存显著差异和选择性。尤其是对人T细胞白血病细胞系MOLT-4、人T淋巴母细胞白血病细胞系Jurkat、小鼠黑色素瘤细胞系B16-F10、人结肠腺癌细胞系SW-480抑制率均>80%,其对应的IC50分别为13.59,24.10,30.05,31.39mg·L-1。结论:THH总生物碱体外具有广泛,高效的抗肿瘤活性,可进一步探讨其作为抗肿瘤药物研究开发的价值。

昆明山海棠片对小鼠淋巴细胞体外增殖活化影响的研究

目的:研究昆明山海棠片对小鼠脾淋巴细胞增殖、活化标志CD69和CD25及细胞因子IL-4及IFN-γ的影响。方法:无菌分离小鼠脾淋巴细胞,加入不同浓度的昆明山海棠溶液,MTT法检测刀豆蛋白A刺激的小鼠脾淋巴细胞增殖;流式细胞仪检测活化标志CD69及CD25;酶联免疫法检测细胞因子IL-4及IFN-γ。结果:昆明山海棠片对脾淋巴细胞增殖活化及分泌均有抑制作用。

昆明山海棠诱导HL-60细胞凋亡的初步研究

目的 探讨中药昆明山海棠 (THH)诱导急性白血病细胞凋亡的规律及发生机制。方法 应用染料排除法、活细胞荧光染色、DNA电泳、流式细胞仪、斑点杂交、原位杂交和荧光 Ca2 +指示剂进行检测和观察。结果 HL-6 0细胞在 THH作用下出现典型的细胞凋亡特征 ,包括细胞形态改变、DNA梯状带以及亚 G1 峰 ,并且存在明显的量效和时效关系。进一步研究表明 ,Bcl- 2基因在凋亡过程中的表达持续下调 ,细胞内游离 Ca2 + 浓度未发生明显变化。结论 THH有较强的诱导白血病 HL- 6 0细胞凋亡的能力 ,其作用机制与下调 Bcl-

基因芯片分析显示昆明山海棠有机萃取液经NF- κB及线粒体信号传导途径诱导HL- 60细胞凋亡(英文)

目的 研究中草药昆明山海棠 Tripterygium hypoglaucum( THH)有机萃取液诱导人早幼粒白血病 HL-6 0细胞凋亡过程的信号传导通道及分子机制。方法 应用流式细胞仪研究了 THH有机萃取液诱导 HL - 6 0细胞的凋亡过程 ,并应用包含 3 0 0 0人类基因与 EST的基因芯片进行基因表达差异分析。结果 基因芯片杂交结果显示有 16个基因表达发生大于 2倍的显著变化 ,这些基因同细胞生长 ,细胞周期调控 ,细胞分化 ,以及压力反应有关。部分基因在细胞凋亡过程中起关键作用 ,如 Caspase3和 Caspase8。结论 THH有机萃取液诱导 HL- 6 0细胞凋亡 ,该过程与 NF- κB和线粒体信号传导途径有关。

长期服用昆明山海棠对雄性大鼠抗生育作用可逆性的研究

成年雄性SD大鼠灌胃给昆明山海棠恨的乙醇提取物(简称ATH),每日0.3g/kg,6周后所有用药鼠均丧失生育力,检测其附睾尾精子活率及密度均明显低于对照。继续给维持量(0.2g/kg)ATH16周,停药5周后生育力及附睾精子完全恢复,所测的其它各项指标包括体重、附性腺重量、睾丸、附睾及主要脏器的组织学、血常规及淋巴细胞转化试验均与对照组无显著性差异。结果表明,在雄性SD 大鼠,低剂量的ATH 长期(22周)给药所致的不育是完全可逆的,且不遗留任何明显毒副作用。

昆明山海棠对血管内皮细胞ICAM-1表达的影响

目的 研究昆明山海棠对猴视网膜血管内皮细胞表达ICAM-1的影响。方法 采用免疫组化测定培养内皮细胞粘附分子表达。结果 ＬＰＳ可使内皮细胞 ＩＣＡＭ-1表达增强，昆明山海棠使 ＩＣＡＭ-1表达减弱。结论 昆明山海棠可通过抑制ＩＣＡＭ-1的表达，降低白细胞与内皮细胞粘附来实现其抗炎机制。

昆明山海棠总生物碱诱导肺腺癌A549细胞凋亡与细胞周期改变

目的为探讨昆明山海棠总生物碱(alkaloids from Tripterygium Hypoglaucum Hutch,THHa)抗肺癌机制,研究THHa诱导肺腺癌A549细胞凋亡与细胞周期改变的关系。方法应用形态学观察、DNA凝胶电泳、流式细胞仪检测分析肺腺癌A549细胞凋亡及细胞周期改变。结果THHa能够诱导肺腺癌A549细胞凋亡并主要作用于细胞周期S期(S期细胞增多)。结论THHa可能通过诱导细胞凋亡而抑制A549细胞增殖,这种作用与细胞周期有关,这些有助于将来肺癌治疗方案的合理设计。