贵州苗药方“四大血”对CIA大鼠滑膜血管增生的作用机制  

The mechanism of Guizhou Miao Yaofang’s Sidaxue on synovial blood vessel growth in CIA rats

作　　者：袁桃花 黄宇键 李佳豪 曹恒 吴昌学[3] 吴宁 YUAN Tao-hua;HUANG Yu-jian;LI Jia-hao;CAO Heng;WU Chang-xue;WU Ning(School of Basic Medical Science, Guizhou Medical University, Guiyang 550025, China;College of Clinical Medicine, Guizhou Medical University, Guiyang 550025, China;Key Laboratory of Molecular Biology, Guizhou Medical University, Guiyang 550025, China)

机构地区：[1]贵州医科大学基础医学院,贵州贵阳550025 [2]贵州医科大学临床医学院,贵州贵阳550025 [3]贵州医科大学医学分子生物学重点实验室,贵州贵阳550025

出　　处：《西安交通大学学报（医学版）》2019年第5期824-829,F0003,共7页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：贵州省科技合作计划项目(黔科合LH字[2015]7330);2014年贵州省卫生计生委科学技术基金项目;贵州省研究生教育教学改革重点课题(No.07100120241)~~

摘　　要：目的研究贵州苗药方“四大血”(SX)对胶原蛋白诱导的关节炎(CIA)大鼠滑膜血管增生的作用机制。方法42只SD大鼠随机分成“四大血”高、中、低剂量组(SX 40g/kg组、SX 20g/kg组、SX 10g/kg组),雷公藤多苷片对照组(GTW)、空白对照组(Nor)、模型组(Mod),每组7只。构建牛Ⅱ型CIA大鼠模型。测量大鼠后肢足跖的肿胀程度并进行关节炎指数(AI)评分,蛋白免疫印记法检测滑膜组织中人第10号染色体缺失的磷酸酶及张力蛋白同源的基因(PTEN)、磷酯酰肌醇3激酶(PI3K)、蛋白激酶B(AKT)的表达,实时荧光定量法检测大鼠滑膜组织中血管内皮生长因子(VEGF)和血管内皮生长因子受体2(VEGFR-2)mRNA表达,酶联免疫吸附试验检测大鼠血清VEGF的表达水平。结果 GTW组及SX各治疗组关节炎指数(AI)评分和足肿度都小于Mod组(P<0.01)。在光镜下观察大鼠足趾病理组织切片,Mod组可见大量炎性细胞浸润,各层组织结构混乱。与Mod组相比,SX组和GTW组炎性细胞浸润明显下降,纤维组织增生水肿减轻,炎性细胞数减少(P<0.01);血清中VEGF含量降低(P<0.01);GTW和SX40g/kg、10g/kg剂量组滑膜组织中VEGF、VEGFR-2mRNA的表达水平均降低(P<0.05,P<0.01);GTW组、SX组关节滑膜组织中PTEN的表达上升(P<0.05,P<0.01);SX组和GTW组AKT、PI3K蛋白表达下调(P<0.01)。结论 SX能上调负调控蛋白PTEN,进而减缓PI3K/AKT通路的活化程度,下调VEGF、VEGFR-2的表达。本研究为RA的治疗提供了新靶点。Objective To study the mechanism of “Sidaxue”(SX) on the synovial vascular proliferation in collagen induced arthritis (CIA) rats.Methods Forty-two SD rats were randomly divided into SX 40 g/kg group, SX 20 g/kg group, SX 10 g/kg group, GTW control group, blank control group (Nor), and model group (Mod).The animal model of arthritis induced by bovine type Ⅱ collagen was constructed.The degree of swelling of the rats hind limbs the was measured and arthritis index (AI) was scored.Western blotting was adopted to detect the expression of phosphatase and tensin homolog deleted on chromosome ten (PTEN), phosphoinositide 3 kinase (PI3K) and protein kinase B (AKT) in synovial tissue.Real-time quantitative RT-PCR was used to detect the expressions of VEGF and VEGFR-2 mRNA in rat synovial tissue.The expression of VEGF protein in serum was detected by enzyme-linked immunosorbent assay.Results The AI score and foot swelling in GTW group and each treatment group of SX were lower than those in the Mod group ( P <0.01).The pathological tissue sections of rat toes were observed under light microscope, and a large number of inflammatory cells infiltrated in the Mod group and the tissue structure of each layer was disordered.Compared with those in the Mod group, the inflammatory cell infiltration and fibrous tissue edema in SX group and GTW group decreased significantly, the number of inflammatory cells decreased ( P <0.01);VEGF level in Serum of decreased ( P <0.01), the expression levels of VEGF mRNA and VEGFR-2 mRNA in synovial tissue of the GTW and SX 40 g/kg and 10 g/kg groups all decreased ( P <0.05, P <0.01), the expression of PTEN increased in synovial tissues of GTW and SX groups ( P <0.05, P <0.01), and the expressions of AKT and PI3K in SX group and GTW group was down-regulated ( P <0.01).Conclusion SX can up-regulate the negative regulatory protein PTEN, thus slowing down the activation of PI3K/AKT pathway and down-regulating the expressions of VEGF and VEGFR-2.This study provides new targets for the treatment o

关 键 词：苗药验方“四大血” 类风湿性关节炎 PTEN/PI3K/AKT通路 

分 类 号：R34[医药卫生—基础医学]