中国中药杂志

2019, v.44(16) 3520-3525

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雷公藤甲素通过调控VEGFA,SDF-1,CXCR4通路改善强直性脊柱炎患者血小板活化的机制
Effect of triptolide in improving platelet activation in patients with ankylosing spondylitis by regulating VEGFA,SDF-1,CXCR4 pathway

方妍妍;万磊;董文哲;文建庭;刘健;
FANG Yan-yan;WAN Lei;DONG Wen-zhe;WEN Jian-ting;LIU Jian;Graduate School of Anhui University of Traditional Chinese Medicine;the First Affiliated Hospital of Anhui University of Traditional Chinese Medicine;

摘要(Abstract):

通过体外实验研究雷公藤甲素(triptolide,TP)对VEGFA,SDF-1,CXCR4通路的影响,探讨其改善强直性脊柱炎(AS)患者血小板活化的机制。外周血单核细胞(PBMC)用于实验。分为4组:正常组(NC)、模型组(MC)、雷公藤甲素组(TP)、AMD3100组。MTT法检测TP最佳浓度。ELISA法检测TNF-α,IL-1β,IL-4,IL-10,VEGFA,VEGFR的表达,实时荧光定量PCR(RT-qPCR)法检测SDF-1,CXCR4,VEGFA的表达,Western blot法检测SDF-1,CXCR4,VEGFA,VEGFR蛋白的表达,免疫荧光法检测CD62p,CD40L,PDGFA的表达水平。MTT结果显示中剂量TP在24 h时对细胞抑制作用最强。ELISA及PCR法结果显示TP可抑制IL-1β,TNF-α,VEGFA,VEGFR,SDF-1,CXCR4,VEGFA mRNA的表达; Western blot结果说明TP可抑制SDF-1,CXCR4,VEGFA,VEGFR蛋白的表达;免疫荧光结果说明TP可抑制CD62p,CD40L,PDGFA的表达。TP可能通过下调SDF-1,CXCR4,VEGFA,VEGFR mRNA,进而下调IL-1β,TNF-α,上调IL-4,IL-10细胞因子的表达,从而调节血小板活化。
The effect of triptolide( TP) on VEGFA,SDF-1,CXCR4 pathway were investigated in vitro to explore the mechanism in improving platelet activation in patients with ankylosing spondylitis( AS). Peripheral blood mononuclear cells( PBMC) were used for the experiment and divided into 4 groups: normal group( NC),model group( MC),triptolide group( TP),and AMD3100 group. The optimal concentration of TP was measured by the MTT method. The expressions of TNF-α,IL-1β,IL-4,IL-10,VEGFA and VEGFR were detected by ELISA. The expressions of SDF-1,CXCR4 and VEGFA were detected by real-time quantitative PCR( RT-qPCR).The expressions of SDF-1,CXCR4,VEGFA and VEGFR were detected by Western blot. The expression levels of CD62 p,CD40 L and PDGFA were detected by immunofluorescence. MTT results showed that medium-dose TP had the strongest inhibitory effect on cells at24 h. The results of ELISA and PCR showed that TP inhibited mRNA expressions of IL-1β,TNF-α,VEGFA,VEGFR and SDF-1,CXCR4 and VEGFA. The results of Western blot indicated that TP inhibited SDF-1,CXCR4 and VEGFA,VEGFR protein expressions; immunofluorescence results indicate that TP can inhibit the expressions of CD62 p,CD40 L,PDGFA. TP may regulate platelet activation by down-regulating SDF-1,CXCR4,VEGFA and VEGFR mRNA expressions,thereby down-regulating IL-1β and TNF-αexpressions,and up-regulating the expressions of IL-4 and IL-10 cytokines.

关键词(KeyWords): 强直性脊柱炎;雷公藤甲素;VEGFA;SDF-1;CXCR4;血小板活化
ankylosing spondylitis;triptolide;VEGFA;SDF-1;CXCR4;platelet activation

Abstract:

Keywords:

基金项目(Foundation): 安徽省科技攻关项目(1604A0802085);; 2016年中央引导地方科技发展专项(财教[2016]1188);; 现代中医内科应用基础与开发研究安徽省重点实验室项目(财教[2016]518);; 2015年技术创新服务体系专项(财教[2015]1551);; 中医药行业科研专项(201307001);; 安徽高校自然科学研究重点项目(KJ2017A281)

作者(Authors): 方妍妍;万磊;董文哲;文建庭;刘健;
FANG Yan-yan;WAN Lei;DONG Wen-zhe;WEN Jian-ting;LIU Jian;Graduate School of Anhui University of Traditional Chinese Medicine;the First Affiliated Hospital of Anhui University of Traditional Chinese Medicine;

DOI: 10.19540/j.cnki.cjcmm.20181113.001

参考文献(References):

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