黄芪桂枝五物汤对骨关节炎大鼠血管新生的作用  

Effect of Huangqi Guizhi Wuwu Tang on Angiogenesis in Osteoarthritis

作　　者：赵乐[1,2] 李艳彦 王永辉 董小君 周然[2] 张若楠 孙力威 ZHAO Le;LI Yan-yan;WANG Yong-hui;DONG Xiao-jun;ZHOU Ran;ZHANG Ruo-nan;SUN Li-wei(Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China;Shanxi University of Chinese Medicine,Taiyuan 030024,China;Zunyi Medical and Pharmaceutical College,Zunyi 563006,China)

机构地区：[1]成都中医药大学,成都611137 [2]山西中医药大学,太原030024 [3]遵义医药高等专科学校,贵州遵义563006

出　　处：《中国实验方剂学杂志》2019年第3期87-93,共7页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：山西省卫生厅项目(201601110);山西省科技厅项目(2014011047-1);山西中医药大学博士科研启动项目

摘　　要：目的:研究黄芪桂枝五物汤对阳虚寒凝骨关节炎(osteoarthritis,OA)模型大鼠血管内皮生长因子(VEGF)及相关因子的影响。方法:60只SPF级SD大鼠随机分为正常组,模型组,塞来昔布组(20.82 mg·kg^-1),黄芪桂枝五物汤低、中、高剂量组(3.24,6.48,12.96 g·kg^-1)。除正常组外,其余各组通过冷固法复合寒冷环境刺激42 d,复制OA模型。造模后,给药组分别灌胃相应药物,正常组和模型组给同等剂量蒸馏水,每天1次,连续28 d。末次给药24 h后,酶联免疫吸附测定(ELISA)检测血清中VEGF,前列腺素E2(PGE2),转化生长因子β1(TGF-β1)水平;免疫组化染色检测左膝关节滑膜和软骨内VEGF表达;实时荧光定量聚合酶链式反应(Real-time PCR)检测右膝关节滑膜白细胞介素-17(IL-17)和VEGF mRNA表达。结果:与正常组比较,模型组血清、软骨和滑膜组织中VEGF表达均显著性升高(P<0.01),同时,血清中PGE2和TGF-β1,滑膜组织IL-17水平均显著升高(P<0.01);与模型组比较,各用药组血清、软骨和滑膜组织中VEGF表达均显著性降低(P<0.05),同时,血清中PGE2和TGF-β1水平,滑膜组织中IL-17水平亦显著降低(P<0.05);各用药组间比较,黄芪桂枝五物汤高剂量组膝关节滑膜组织IL-17较塞来昔布组升高(P<0.05),黄芪桂枝五物汤中、高剂量组VEGF较塞来昔布组显著升高(P<0.01),其余各组间比较均无显著性差异。结论:黄芪桂枝五物汤可能通过减轻骨关节炎大鼠血管新生相关细胞因子PGE2和TGF-β1等的表达而作用于VEGF,从而抑制膝骨关节处血管新生,减轻软骨损伤。Objective:To study the effect of Huangqi Guizhi Wuwu Tang on angiogenesis of osteoarthritis with Yang deficiency and cold coagulation.Method:Totally 60 female SD rats were randomly divided into control group,model group,celecoxib group(20.82 mg·kg^-1)and low,medium,high-dose Huangqi Guizhi Wuwu Tang groups(3.24,6.48,12.96 g·kg^-1).All groups,except for control group,were involved in duplicating the osteoarthritis(OA)model through frozen and knee fixation,as well as 42-day cold environmental stimulation.After modeling,all drug-group rats were respectively administrated with corresponding drugs for 28 days,once a day.Meanwhile,control group and model group were given equivalent distilled water by gavage.24 hours after the last gavage,vascular endothelial growth factor(VEGF),prostaglandin E2(PGE2)and transforming growth factor-β1(TGF-β1)in serum were detected with enzyme linked immunosorbent assay(ELISA)method,VEGF expressions in cartilage and synovial with immunohistochemical method,and interleukin-17(IL-17)and VEGF levels in synovial with Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR).Result:Compared with normal group,the expression of VEGF in serum,cartilage and synovial were significantly increased(P<0.01),and PGE2and TGF-β1expressions,IL-17 level were increased significantly(P<0.01)as well.Compared with model group,VEGF in serum,cartilage and synovial were significantly decreased in the 4 drug groups.Meanwhile,PGE2,TGF-β1and IL-17 level were decreased significantly(P<0.05).Among drug groups,IL-17 of high-dose group in synovial was higher than that of celecoxib group(P<0.05),and VEGF of medium and high-dose Huangqi Guizhi Wuwu Tang groups in synovial was higher than that of celecoxib group(P<0.05).There was no significant difference among other drug groups.Conclusion:Huangqi Guizhi Wuwu Tang has an effect in suppressing angiogenesis of knee and alleviate cartilage lesion by regulating VEGF and its upstream cytokines PGE2and TGF-β1.

关 键 词：黄芪桂枝五物汤 骨关节炎 血管内皮生长因子(VEGF) 血管新生 

分 类 号：R20[医药卫生—中医学] R22