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.

ANEUPLOIDY INDUCTION BY THE WATER EXTRACT OF TRIPTERYGIUM HYPOGLAUCUM(LEVEL) HUTCH IN MOUSE BONE MARROW CELLS

The aneuploldy inducing activity of a Chinese medicinal herb, Tripterygium hypoglaucum (level) Hutch (THH) were investigated by means of thhree cytogenetic end points, namely C-mitotic (CM) effects, micronucleus (MN) and parallel chromosome structural aberration (CA) analyses in vivo. The experiments were performed on mouse bone marrow cells. THH showed similar gentoxic effects to colchicine (COL) in CM, MN and CA analyses; positive CM effects were observed accompanied with increases of mitotic index and frequencies of C-mltotic cells as well as decreased frequencies of anaphase in all of the THH-treated groups. The compound showed a positive MN response in bone marrow polychromatic erythrocytes but was negative in CA analyses. The preliminary results suggested that THH is an aneuploldy inducer in mouse bone marrow cells under present experiment conditions.

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.

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。

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.

Identification of a cytochrome P450 from Tripterygium hypoglaucum(Levl.)Hutch that catalyzes polpunonic acid formation in celastrol biosynthesis

Tripterygium hypoglaucum(Levl.)Hutch,a traditional Chinese medicinal herb with a long history of use,is widely distributed in China.One of its main active components,celastrol,has great potential to be developed into anti-cancer and anti-obesity drugs.Although it exhibits strong pharmacological activities,there is a lack of sustainable sources of celastrol and its derivatives,making it crucial to develop novel sources of these drugs through synthetic biology.The key step in the biosynthesis of celastrol is considered to be the cyclization of 2,3-oxidosqualene into friedelin under the catalysis of 2,3-oxidosqualene cyclases.Friedelin was speculated to be oxidized into celastrol by cytochrome P450 oxidases(CYP450s).Here,we reported a cytochrome P450 ThCYP712K1 from Tripterygium hypoglaucum(Levl.)Hutch that catalyzed the oxidation of friedelin into polpuonic acid when heterologously expressed in yeast.Through substrate supplementation and in vitro enzyme analysis,ThCYP712K1 was further proven to catalyze the oxidation of friedelin at the C-29 position to produce polpunonic acid,which is considered a vital step in the biosynthesis of celastrol,and will lay a foundation for further analysis of its biosynthetic pathway.

A New Natural Compound with Cytotoxic Activity from Tripterygium hypoglaucum

Objective To find antitumor constituents from Tripterygium hypoglaucum.Methods The chloroform extract of T.hypoglaucum was separated by silica gel column chromatography and preparative HPLC.The structures of compounds isolated were identified by spectral analysis and chemical evidence.Results Seven compounds were isolated and identified as rhein ethyl ester(1),chrysophenol(2),physcion(3),emodin(4),wilfordine(5),wilforgine(6),and wilforine(7).The cytotoxic activities of the compounds against cancer cell lines were assayed.Conclusion Compound 1 is a new natural compound with strong activities against human cancer cell lines(A2780 and OVCAR-3) .Compounds 2-4 are isolated from this genus plants for the first time.The possible structure-activity relationship among compounds 1-4 shows that the methoxy group or oxyethyl moiety might be responsible for the cytotoxity.

The Effects of Astragalus membranaceus and Tripterygium hypoglaucum on NK Activity in Systemic Lupus Erythematosus

The natural killer cell activity of peripheral blood mononuclear cells from 28 patients withsystemic lupus erythematosus was measured using enzyme-release assay. The patients exhibited obvi-ously decreased natural killer cell activity compared with the healthy subjects in the control group. Thenatural killer cell dysfonction was related to steroidal therapy, but not to three auto-antibodies. The re-lease of a soluble natural killer cytotoxic factor by the peripheral blood mononuclear cells was tested bycytotoxicity assay induced in K256 cells. This release was significantly lower in the test group than in thecontrol group. Pre-incubation of peripheral blood mononuclear cells with Astragalus membranaceus andTripterygium hypoglaucum considerably enhanced natural killer cytotoxicity by stimulating the release ofsoluble cytotoxic factors.

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。

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。

雷公藤属3种植物遗传关系与遗传多样性的RAPD分析

目的:阐明雷公藤属植物的遗传关系和遗传多样性。方法:用RAPD分子标记方法研究来自雷公藤属植物3个种4个类型24个居群的110个样本。结果和讨论:从100个随机引物中筛选出10个引物,得到128条带,其中多样性条带123个,占96%。通过聚类分析可以看出,东北雷公藤明显与其他种有显著的遗传差异;中间类型和雷公藤的关系较与昆明山海棠的关系更密切,但中间类型和雷公藤之间也有明显的遗传分化,且中间类型呈现较雷公藤复杂的遗传多样性,不同的中间类型居群与雷公藤的关系远近不同。大量的中间类型的存在,连接了昆明山海棠与雷公藤这2个种,从分类上建议将2个种合并。将除去东北雷公藤的其他居群作为一个整体分析,居群间分化明显,遗传多样性主要存在于居群间,因此应当收集不同来源的种质以保存尽可能多的遗传多样性。

狭叶崖爬藤化学成分的研究(英文)

目的 研究我国特有药用植物狭叶崖爬藤 Tetrastigma hypoglaucum的化学成分 ,为阐明其有效成分提供依据。方法 利用各种色谱技术进行分离 ,根据化合物的光谱数据 (IR,MS,1 HNMR,1 3CNMR)和化学方法鉴定其结构。结果 从狭叶崖爬藤地上部分分离并鉴定了 10个化合物 ,分别为β-谷甾醇 (β- sitosterol, ) ,棕榈酸(palm itic acid, ) ,正二十五烷 (pentacosane, ) ,胡萝卜苷 (daucosterol, ) ,白藜芦醇 (resveratrol, ) ,没食子酸(gallic acid, ) ,没食子酸乙酯 (ethyl gallate, ) ,儿茶素 (catechin, ) ,7-氧 -没食子酰基 -儿茶素 (7- O- galloylcate-chin, )和 3,3′-二甲氧基鞣花酸 - 4 -氧 -葡萄糖苷 (3,3′- dimethoxy ellagic acid- 4 - O- β- D- glucopyranoside, )。结论所有化合物均系首次从该植物中分离得到。

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。