丹参酮ⅡA对腹膜透析液诱导的人腹膜间皮细胞转分化及致纤维化因子表达的影响  

Effects of Tanshinone ⅡA on epithelial-myofibroblast transdifferentiation and fibrogenic cytokine expression of human peritoneal mesothelial cells induced by peritoneal dialysis fluid

作　　者：邵秋媛[1] 蒋春明[1] 孙琤[1] 于立杰[1] 张苗[1] 

机构地区：[1]南京大学医学院附属鼓楼医院肾脏科

出　　处：《医学研究生学报》2012年第1期20-25,共6页Journal of Medical Postgraduates

基　　金：南京市重点科技发展项目(ZKX08026)

摘　　要：目的腹膜纤维化是长期腹膜透析(peritoneal dialysis,PD)的常见并发症,也是透析不充分和退出PD的主要原因,研究表明丹参酮ⅡA能改善各种组织中的纤维化。文中研究丹参酮ⅡA磺酸钠注射液对在高糖腹膜透析液(peritonealdialysis fluid,PDF)诱导下体外培养的人腹膜间皮细胞(human peritoneal mesothelial cells,HPMCs)纤维化及其相关因子表达的影响。方法选择非尿毒症、非糖尿病择期手术患者的大网膜作为HPMCs的来源,将培养的细胞随机分成4组:对照组、PDF组、丹参酮1组(含丹参酮ⅡA浓度为50μmol/L的PDF组)、丹参酮2组(含丹参酮ⅡA浓度为100μmol/L的PDF组),同步培养72h后,应用间接免疫荧光法检测α平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、上皮钙黏素(E-cadherin)表达,实时荧光定量PCR检测E-cadherin、α-SMA、纤维连接蛋白(fibronectin,FN)、Ⅰ型胶原酶(CollagenⅠ)、转化生长因子-β1(transforming growth factor-β1,TGF-β1)、smad2、smad7 mRNA表达。结果免疫荧光法发现,丹参酮ⅡA能显著抑制高糖诱导的E-cadherin低表达和α-SMA高表达。Real-time-PCR发现,添加丹参酮ⅡA的PDF组α-SMA、FN、CollagenⅠ、TGF-β1、smad2 mRNA表达与PDF组比较显著下调(P<0.05);E-cadherin、smad7 mRNA表达显著上调(P<0.05)。结论在PDF中添加丹参酮ⅡA可显著减轻PDF导致的HPMCs转分化,从而抑制纤维化相关因子的表达。Objective Peritoneal fibrosis is a common complication of long-term peritoneal dialysis（PD） as well as the main cause of dialysis inadequacy and PD withdrawal.It has been reported that Tanshinone ⅡA can ameliorate fibrosis in various tissues.The aim of this study is to investigate the effects of Tanshinone ⅡA on the fibrosis of human peritoneal mesothelial cells（HPMCs） induced by PD fluid（PDF） and the expressions of fibrogenic cytokines.Methods HPMCs were cultured from the omentum of the patients undergoing selective abdominal surgery without uremia and diabetes,and then divided into four groups： control,PDF,Tanshinone 1（50 μmol/L Tanshinone ⅡA）,and Tanshinone 2（100 μmol/L Tanshinone ⅡA）.After 72 hours of treatment,the expressions of α-smooth muscle actin（α-SMA） and E-cadherin were measured by indirect immunofluorescent assay,and the expressions of α-SMA,E-cadherin,FN,Collagen Ⅰ,TGF-β1,smad2,and smad7 mRNA detected by real-time PCR.Results Tanshinone ⅡA markedly inhibited the PDF-induced downregulation of E-cadherin and upregulation of α-SMA expression.Compared with PDF alone,PDF with Tanshinone ⅡA significantly decreased the expressions of α-SMA,FN,Collagen Ⅰ,TGF-β1 and smad2 mRNA（P0.05）,and increased the expressions of E-cadherin and smad7 mRNA（P0.05）.Conclusion Addition of Tanshinone ⅡA to PDF can significantly counteract PDF-mediated transdifferentiation of HPMCs,and thus inhibit the expressions of fibrogenic cytokines.

关 键 词：丹参酮ⅡA 腹膜透析液 人腹膜间皮细胞 腹膜纤维化 间叶转分化 

分 类 号：R459.5[医药卫生—治疗学]