中华小儿外科杂志
中華小兒外科雜誌
중화소인외과잡지
CHINESE JOURNAL OF PEDIATRIC SURGERY
2008年
4期
242-245
,共4页
丹参酮ⅡA%成纤维细胞%转化生长因子β%Smad蛋白质类
丹參酮ⅡA%成纖維細胞%轉化生長因子β%Smad蛋白質類
단삼동ⅡA%성섬유세포%전화생장인자β%Smad단백질류
Tanshinone Ⅱ A%Fibroblasts%Transforming growth factor beta%Smad proteins
目的 研究丹参酮Ⅱ A(tanshinone Ⅱ A,TSN)对大鼠.肾间质成纤维细胞转化生长因子β1(transforming growth factor β1,TGFlβ1)信号转导的影响,探讨TSN抑制肾间质纤维化的作用机制.方法 体外培养大鼠肾成纤维细胞株NRK/49F,应用5 ng/ml TGFβ1刺激及不同浓度(10-6、10-5、10-4 mol/L)TSN预处理.逆转录聚合酶链反应(reverse transcriptase polymerase chain reaction,RT-PCR)检测纤维连接蛋白(fibronectin,FN)mRNA表达,Western blot检测FN和Smads蛋白表达.结果 TGFβ1在一定范围内以时间依赖方式诱导FN及Smads表达,刺激终末FN mRNA和蛋白表达量分别增加1.1倍和1.5倍(P＜0.01,P＜0.01),磷酸化Smad2/3(phosphorylated Smad2/3,p-Smad2/3)蛋白表达量增加7倍(P＜0.01).TSN预处理可下调FN和p-Srnad2/3表达,而且效应呈剂量依赖性.10-6mol/L TSN预处理对FN和p-Srnad2/3表达几乎无影响(P＞0.05,P＞0.05);10-5和10-4mol/LTSN预处理后,FNmRNA表达量分别下降28.1%、43.8%(P＜0.05,P＜0.01),FN蛋白表达量分别下降40%、44%(P＜0.05,P＜0.05),p-Smad2/3蛋白表达量分别下降40%、65%(P＜0.05,P＜0.01).结论 TSN抗肾间质纤维化作用可能与其阻断.肾间质成纤维细胞TGFβ1-Srnads信号通路有关.
目的 研究丹參酮Ⅱ A(tanshinone Ⅱ A,TSN)對大鼠.腎間質成纖維細胞轉化生長因子β1(transforming growth factor β1,TGFlβ1)信號轉導的影響,探討TSN抑製腎間質纖維化的作用機製.方法 體外培養大鼠腎成纖維細胞株NRK/49F,應用5 ng/ml TGFβ1刺激及不同濃度(10-6、10-5、10-4 mol/L)TSN預處理.逆轉錄聚閤酶鏈反應(reverse transcriptase polymerase chain reaction,RT-PCR)檢測纖維連接蛋白(fibronectin,FN)mRNA錶達,Western blot檢測FN和Smads蛋白錶達.結果 TGFβ1在一定範圍內以時間依賴方式誘導FN及Smads錶達,刺激終末FN mRNA和蛋白錶達量分彆增加1.1倍和1.5倍(P＜0.01,P＜0.01),燐痠化Smad2/3(phosphorylated Smad2/3,p-Smad2/3)蛋白錶達量增加7倍(P＜0.01).TSN預處理可下調FN和p-Srnad2/3錶達,而且效應呈劑量依賴性.10-6mol/L TSN預處理對FN和p-Srnad2/3錶達幾乎無影響(P＞0.05,P＞0.05);10-5和10-4mol/LTSN預處理後,FNmRNA錶達量分彆下降28.1%、43.8%(P＜0.05,P＜0.01),FN蛋白錶達量分彆下降40%、44%(P＜0.05,P＜0.05),p-Smad2/3蛋白錶達量分彆下降40%、65%(P＜0.05,P＜0.01).結論 TSN抗腎間質纖維化作用可能與其阻斷.腎間質成纖維細胞TGFβ1-Srnads信號通路有關.
목적 연구단삼동Ⅱ A(tanshinone Ⅱ A,TSN)대대서.신간질성섬유세포전화생장인자β1(transforming growth factor β1,TGFlβ1)신호전도적영향,탐토TSN억제신간질섬유화적작용궤제.방법 체외배양대서신성섬유세포주NRK/49F,응용5 ng/ml TGFβ1자격급불동농도(10-6、10-5、10-4 mol/L)TSN예처리.역전록취합매련반응(reverse transcriptase polymerase chain reaction,RT-PCR)검측섬유련접단백(fibronectin,FN)mRNA표체,Western blot검측FN화Smads단백표체.결과 TGFβ1재일정범위내이시간의뢰방식유도FN급Smads표체,자격종말FN mRNA화단백표체량분별증가1.1배화1.5배(P＜0.01,P＜0.01),린산화Smad2/3(phosphorylated Smad2/3,p-Smad2/3)단백표체량증가7배(P＜0.01).TSN예처리가하조FN화p-Srnad2/3표체,이차효응정제량의뢰성.10-6mol/L TSN예처리대FN화p-Srnad2/3표체궤호무영향(P＞0.05,P＞0.05);10-5화10-4mol/LTSN예처리후,FNmRNA표체량분별하강28.1%、43.8%(P＜0.05,P＜0.01),FN단백표체량분별하강40%、44%(P＜0.05,P＜0.05),p-Smad2/3단백표체량분별하강40%、65%(P＜0.05,P＜0.01).결론 TSN항신간질섬유화작용가능여기조단.신간질성섬유세포TGFβ1-Srnads신호통로유관.
Objective To study the effects of tanshinone Ⅱ A(TSN)on transforming growth factor 131(TGFβ1)signal transduction in renal interstitial fibroblasts of rats and to investigate its mechanism in prevention of tubulointerstmal fibrosis.Methods Rat renal fibroblasts of the line NRK/49F were cultured in vitro,stimulated with 5ng/mL TGFβ1 and pretreated with 10-6,10-5,and 10-4mol/L TSN,respectively.The mRNA levels of fibronectin(FN)were analyzed by reverse transcriptase polymerase chain reaction(RT-PCR).The protein expression of FN and Srnads were detected by western blot.Results TGFβ1 induced the expression of FN and Smads in a time-dependent manner.Compared with pro-stimulation,the FN mRNA and protein levels increaseel by 1.1 times and 1.5 times respectively(P＜ 0.01),the protein expression of phosphorylated Smad2/3(p-Smad2/3)increflsed by 7 times at the end of TGFβ1 stimulation(P＜0.01).TSN precondition down-regulated FN and p-Smad2/3 expression in a dose-dependent nlannero 10-6mol/L TSN precondition had no effect on FN and p-Smad2/3 expression(P＞0.05).After pretreated with 10-and 10-4mol/L TSN,the FN mRNA levels decreased by 28.1%and 43.8%(P＜0.05 and P＜0.01),the FN protein levels decreased by 40%and 44%(P＜0.05),and the p-Smad2/3 protein expression decreased by 40%and 65%respectively(P＜0.05 and P＜0.01).Conclusions TSN can block TGFβ1-Smads signaling pathway in renal interstitial fibroblasts to inhibit renaIinterstitial fibrosis.
