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机构地区:[1]宜昌卫生学校附属医院 [2]华中科技大学同济医学院附属同济医院,武汉430030
出 处:《内科急危重症杂志》2008年第5期242-245,共4页Journal of Critical Care In Internal Medicine
基 金:国家自然科学基金资助项目(No:30571645)
摘 要:目的:研究丹参酮ⅡA(TSN)对大鼠心脏成纤维细胞(CF)内转化生长因子β1(TGFβ1)信号转导的影响,探讨TSN抗心肌纤维化的作用机制。方法:采用胰酶消化法和差速贴壁法获取新生SD大鼠CF,应用5ng/mlTGFβ1刺激及不同浓度(10-6、10-5、10-4mol/L)TSN预处理。逆转录聚合酶链反应(RT-PCR)检测纤维连接蛋白(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1.3倍和1.8倍(P<0.01,P<0.01),磷酸化Smad2/3(p-Smad2/3)蛋白表达量增加3.9倍(P<0.01)。TSN预处理可下调FN和p-Smad2/3表达,而且效应呈剂量依赖性。10-6mol/L TSN预处理对FN和p-Smad2/3表达几无影响(P>0.05,P>0.05);10-5和10-4mol/LTSN预处理后,FN mRNA表达量分别下降32.7%、42.9%(P<0.05,P<0.01),FN蛋白表达量分别下降45.8%、57.1%(P<0.01,P<0.01),p-Smad2/3蛋白表达量分别下降40.5%、55.4%(P<0.05,P<0.01)。结论:TSN抗心肌纤维化作用可能与其抑制TGFβ1诱导的Smad2/3磷酸化,阻断CF内TGFβ1/Smads信号通路有关。Objective: To study the effects of tanshinone ⅡA(TSN) on signal transduction of transforming growth factor t31(TGFβ1) in cardiac fibroblasts (CF) of rats so as to investigate its mechanism in prevention of myocardial fibrosis. Methods: CF of newborn Sprague-Dawley(SD) rats were isolated by trypsin digestion and differen tial adhesion method, stimulated with 5 ng/mL TGFβ1 and pretreated with 10^- 6, 10^-5, 10^- 4 mol/L TSN respectively. The mRNA level of fibronectin (FN) was determined with RT-PCR. The protein expression of FN and Smads were detected with 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 increased by 1.3 times and 1.8 times respectively (P〈0. 01,P〈0. 01), the protein expression of phosphorylated Smad2/3 (p-Smad2/3) increased by 3.9 times at the end of TGFβ1 stimulation (P〈0. 01). TSN preconditioning down-regulated FN and p-Smad2/3 expression in a dose-dependent manner. 10^-6 mol/L TSN preconditioning had no effect on FN and p-Smad2/3 expression (P) 0. 05,P〉0. 05). After pretreated with 10^-5 and 10 4mol/L TSN, the FN mRNA levels decreased by 32. 7% and 42. 9% (P〈0. 05 ,P〈0. 01), the FN protein levels decreased by 45.8% and 57. 1% (P〈0. 01, P〈0. 01 ), and the p-Smad2/3 protein expression decreased by 40. 5% and 55.4% respectively (P〈0. 05,P〈0. 01). Conclusion: TSN can block TGFβ1/Smads signaling pathway in CF by inhibiting TGFβ1 induced Smad2/3 phosphorylation to prevent myocardial fibrosis.
关 键 词:丹参酮ⅡA转化生长因子81 Smads成纤维细胞
分 类 号:R542.23[医药卫生—心血管疾病]
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