三七总苷对尿毒血清诱导的人肾小管上皮细胞TGF-β_1、CTGF基因表达和蛋白分泌的影响  

The protection effects of Panax Notoginsenosides on the gene expressionand protein excretion of TGF-β_1 and CTGF of human renal tubular epithelial cell induced by uremic serum

作　　者：刘海燕[1] 陈孝文[1] 刘华锋[1] 梁东[1] 唐德燊[1] 

机构地区：[1]广东医学院附属医院肾脏病研究所,广东湛江524001

出　　处：《中国药理学通报》2005年第11期1366-1370,共5页Chinese Pharmacological Bulletin

基　　金：广东省科技计划资助项目(No2003C32702);湛江市科技攻关资助项目(No2003)

摘　　要：目的探讨三七总苷(PNS)对尿毒血清诱导的人肾小管上皮细胞转化生长因子-β1(TGF-β1)、结缔组织生长因子(CTGF)基因表达和蛋白分泌的影响,为临床上应用PNS延缓肾衰竭的进展提供理论依据.方法无菌条件下收集40份尿毒症病人血清和20例正常人血清.应用相差显微镜、扫描电镜结合细胞角蛋白18(CK-18)免疫组化鉴定人肾小管上皮细胞株(HK-2).应用ELISA方法检测HK-2细胞培养上清液TGF-β1蛋白分泌量;Western Blot方法检测CTGF的蛋白表达;RT-PCR方法检测TGF-β1、CTGFmRNA表达.结果 10%尿毒血清组与正常对照组相比,TGF-β1、CTGF基因表达和蛋白分泌量均增高(P<0.01),而PNS 400～800 mg·L-1可恢复尿毒血清所诱导的TGF-β1基因表达(P<0.01),200～800 mg·L-1可恢复尿毒血清诱导的TGF-β1蛋白分泌(P<0.01);200～800 mg·L-1可恢复尿毒血清所诱导的CTGF基因表达和蛋白分泌(P<0.01).结论在体外实验中,PNS可在一定程度上降低尿毒血清诱导的HK-2细胞TGF-β1、CTGF基因表达和蛋白分泌量,减少细胞外基质聚集,从而延缓人肾小管-间质纤维化的进展,有望成为一种能有效延缓肾纤维化进展的中药.Aim To investigate the effects of Panax Notoginsenosides （PNS） on the gene expression and protein excretion of TGF-β1 and CTGF of human renal tubular epithelial cell induced bv uremic serum in vitro. Methods Forty sera from CRF patients and twenty sera from healthy volunteers were collected, gently mixed,inactived and separated respectively in sterfled condition. HK-2 Cells were cultured in RPMI- 1640 medium with 10% newborn calves serum and subcuhured routinely. They were differentiated by phase contrast microscope and seanning electron microscope detection and cytokeratinl8（CK-18） immunohistochemistry method. The protein levels of TGF-β1 were examined by enzyme-linked immunoadsordent assay （ELISA）. The protein levels of CTGF were examined by Western Bloting assay. The gene expression of TGF-β1 and CTGF was detected by Semi-quantitative reverse trainscriptase polymerase chain reaction （RT-PCR）.Results TGF-β1 and CTGF gene expression and protein level were increased in 10% uremic serum groups compared with that of normal control group （P 〈 0. 01 ）,TGF-β1 gene expression and protein secretion induced by uremic serum were decreased in parallel with the increasing of PNS concentration in 400 -800 mg· L^-1 PNS group（P 〈0.01）. CTGF gene and protein expression induced by uremic serum was decreased following the increasing of PNS concentration in 200 800 mg·L^-1 PNS group （ P 〈 0. 01 ）. Conclusion Human renal tubulointerstitial fibrosis can be ameliorated effectively by PNS through inhibiting TGF-β1 and CTGF gene expression and protein secretion induced by uremic toxin in vitro.

关 键 词：三七总苷 尿毒血清 转化生长因子-β1 结缔组织生长因子 

分 类 号：R282.71[医药卫生—中药学] R284.1[医药卫生—中医学]