穿山龙总皂苷对大鼠滑膜细胞株VEGF与AP-1的影响  

Effects of Total Saponins from Rhizoma Dioscreae Nipponicae on VEGF and AP-1 in Rat Synovial Cell Strain

作　　者：高亚贤[1] 王永为[2] 郭亚春[3] 宋鸿儒[1] 肖丽君[1] 安高[1] 梁秀军[1] 翟泽玲[1] 段一娜[4] 

机构地区：[1]承德医学院免疫学教研室,承德067000 [2]承德医学院人体解剖学教研室,承德067000 [3]承德医学院病原生物学教研室,承德067000 [4]承德医学院预防医学教研室,承德067000

出　　处：《医药导报》2015年第3期285-289,共5页Herald of Medicine

基　　金：国家自然科学基金资助项目(30873420);河北省自然科学基金重点项目(C2007000916);河北省高等学校科学技术研究项目(ZH200806)

摘　　要：目的观察含穿山龙总皂苷血清对大鼠滑膜细胞株RSC-364血管内皮细胞生长因子(VEGF)mRNA和激活蛋白-1(AP-1)活性的影响,探讨其抑制血管新生的作用机制。方法制备含穿山龙总皂苷和雷公藤(阳性对照)血清。将培养好的大鼠滑膜细胞株RSC-364分为空白对照组、模型对照组、含雷公藤多苷血清组和含穿山龙总皂苷血清组,培养1 h,除空白对照组外,其余各组加入IL-17和TNF-α(均为10μg·L-1)共同孵育24 h。采用实时荧光定量PCR方法检测各组细胞VEGF mRNA表达,凝胶电泳迁移率实验法(EMSA)检测各组细胞核蛋白提取物AP-1的DNA结合活性。结果与空白对照组比较,模型对照组VEGF mRNA表达水平及AP-1 DNA结合活性显著增高(P<0.05,P<0.01);与模型对照组比较,含雷公藤多苷血清组、含穿山龙总皂苷血清组VEGF mRNA表达水平及AP-1 DNA结合活性均降低(P<0.05),且两组间比较,差异无统计学意义。结论含穿山龙总皂苷血清可以抑制VEGF mRNA表达水平及AP-1 DNA结合活性,其机制可能是通过抑制转录因子AP-1来调控血管新生关键因子VEGF产生,进而抑制血管新生。Objective To study the effects of medicated serum with total saponins from Rhizoma Dioscreae Nipponicae （RDN） on VEGF mRNA expression and AP-1 activity in rat synovial cell strain RSC-364 induced by IL-17 and TNF-α. To investigate the mechanism about total saponin from RDN inhibition of angiogenesis. Methods Medicated serum of total saponins from RDN and tripterygium （positive control） were prepared. Rat synovial cells RSC-364 were divided into four groups： the blank control,IL-17＋TNF-α model,tripterygium medicated serum,and total saponins medicated serum groups. After one hour of incubation,all groups except for the blank control were incubated with both IL-17（10 μg·L^-1 ） and TNF-α（10 μg·L^-1 ） for 24 hours. VEGF mRNA expression in RSC-364 was detected by PrimeScriptTM real-time quantitative PCR （RT-PCR） detection kit,and the AP-1 DNA-binding activity was detected by electrophoretic mobility shift assay （EMSA）. Results Compared with the control blank group,both of the VEGF mRNA expression and AP-1 activity in rat synovial cell strain RSC-364 induced by IL-17 and TNF-α increased remarkably （P〈0. 05,P〈0.01）. The VEGF mRNA expression and AP-1 activity in tripterygium medicated serum group and total saponins medicated serum group were remarkably lower than those of the model control group （P〈0.05）. There was no significant difference between the two medicated serum groups. Conclusion Serum medicated with total saponins from RDN can remarkably decrease VEGF mRNA expression and AP-1 activity,indicating that the total saponins from RDN could influence VEGF secretion by inhibiting the AP-1 signal transduction pathway,VEGF is the key factor of angiogenesis,thereby to restrain angiogenesis.

关 键 词：穿山龙总皂苷 关节炎 类风湿 血管新生 RSC-364 血管内皮生长因子 激活蛋白-1 

分 类 号：R286[医药卫生—中药学] R965[医药卫生—中医学]