银杏叶提取物在TGF-β1诱导的人肾小管上皮细胞表型转化中的作用研究

目的：揭示银杏叶提取物（EGb）在转化生长因子-β1（TGF-β1）诱导的人肾小管上皮细胞（HKC）表型转化中的作用,并探讨其对单核细胞趋化蛋白-1（MCP-1）表达的影响。方法：体外培养HKC,经TGF-β1、EGb干预后,应用细胞免疫组化ABC法检测细胞角化蛋白-18与α-平滑肌肌动蛋白（α-SMA）表达,应用实时定量RT-PCR及Western blot。检测Ⅰ型胶原蛋白,应用Western blot方法检测MCP-1表达。结果：TGF-β1（8ng/ml）刺激HKC细胞72h后,细胞角化蛋白-18表达明显下调,α-SMA、Ⅰ型胶原、MCP-1表达明显上调;加入EGb25、50、100、150mg/L干预72h后,α-SMA、Ⅰ型胶原和MCP-1表达呈剂量依赖性减少,细胞角化蛋白-18表达呈剂量依赖性增加。结论：EGb以浓度依赖性方式抑制TGF-β1诱导的HKC表型转化,该作用机制可能与EGb下调MCP-1基因表达有关。

Detection of the Role of EGb on TGF-β1Induced HKC Epithelial-myofibroblast Transition

Objective： To discuss the role of EGb on TGF- β1 induced HKC epithelial - myofibroblast transition and the effect of EGb on MCP - 1 expression. Methods：Cultured HKC, treated with TGF- β1 ,EGb,we detect the expression of eytokeratin- 18 and α-SMA by cell immunoehemistry ABC,and collagen type Ⅰ by Real-time RT-PCR and Western blot,and MCP- 1 by Western blot. Results：Treated with TGF-β1 （8 ng/ml）72 h, expression of eytokeratin- 18 was down regulated markedly, and expression of α- SMA and collagen type Ⅰ and MCP - 1 were up rgulated markedly. Treated with EGb 25,50,100,150 mg/L and TGF- β1 （8 ng/ml）concomitantly 72 h,expression of 6- SMA and collagen type Ⅰ and MCP- 1 were down regulated in concentration dependent manner, and expression of cytokeratin- 18 was up regulated in concentration dependent manner. Conelusion： EGb can inhibit TGF- β1 induced HKC epithelial- myofibroblast transition in concentration dependent, and the mechanism maybe involve that EGb can down regulate the expression of MCP- 1.