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| 1 | 牛磺酸抑制缺氧诱导大鼠肺动脉平滑肌细胞增殖及信号转导机制显示文摘目的:探讨牛磺酸(taurine,Tau)对缺氧诱导的大鼠肺动脉平滑肌细胞(PASMCs)增殖的影响,并且研究细胞外信号调节激酶1/2(ERK1/2)通路是否参与了Tau抑制PASMCs增殖的过程及可能的分子机制。方法:原代培养大鼠PASMCs,选用第2~5代用于实验。Tau给药浓度为80 mmol·L-1,ERK1/2阻断剂(PD98059)浓度为50μmol·L-1,给药时间24 h。①体外噻唑蓝比色法(MTT)、细胞免疫荧光和蛋白免疫印记法检测牛磺酸在不同条件下对大鼠PASMCs增殖能力的影响,将细胞分为4组:常氧组、常氧+Tau组、缺氧组、缺氧+Tau组。②蛋白免疫印记法检测ERK1/2通路是否参与了Tau抑制缺氧诱导PASMCs增殖的进程中。实验分为5组:常氧组、缺氧组、缺氧+Tau组、缺氧+Tau+PD98059组、缺氧+PD98059组。结果:①缺氧可诱导PASMCs增殖,常氧情况下Tau对PASMCs细胞增殖无影响,常氧及缺氧条件下Tau对PASMCs中肿瘤坏死因子-α(TNF-α)的表达无影响。Tau可以逆转缺氧诱导的细胞增殖核抗原(PCNA)(P〈0.01)、细胞周期蛋白A(Cyclin A)表达上调(P〈0.05)。②常氧情况下Tau对磷酸化细胞外信号调节激酶1/2(p-ERK1/2)的表达没有影响,缺氧使p-ERK1/2的表达上调(P〈0.01),Tau逆转缺氧诱导的p-ERK1/2表达上调(P〈0.05)。PD98059与Tau均可抑制缺氧诱导的PCNA,Cyclin A,p-ERK1/2表达上调,与缺氧单独加药Tau或缺氧单独加入PD98059对比,缺氧+Tau+PD98059组PCNA,Cyclin A,p-ERK1/2表达下调更显著。结论:Tau可抑制缺氧诱导的PASMCs增殖,并且可能是通过ERK1/2通路调控的。 | 张晓丹 孙鹏 朱大岭 谢楠 | 2014 | 中国中药杂志2014，39，10： | 1 |
| 2 | Role of oxygen free radicals in the proliferation of myofibroblasts induced by AngII显示文摘Previous studies have demonstrated the important role of angiotension II(AngII)in promoting proliferation of myofibroblasts(myoFbs)and myocardial fibrosis.However,the underlying mechanisms and the role of oxygen free radicals in the proliferation of myofibroblasts induced by AngII are unclear.The present study was designed to shed light on this issue through exploration of AngII signaling pathways via in vitro experiments.Primary cultures of neonatal rat myoFbs were divided into five groups which were treated with AngII(10^(-8) to 10^(-6) M),AngII with the antioxidant N-acetyl-L-cysteine(NAC),or normal culture medium.We observed the proliferation of myoFbs as induced by AngII at different concentrations with MTT.Reactive oxygen species(ROS)levels in myoFbs were detected by monitoring the fluorescence of 2',7'-dichlorofluorescein.The contents and levels of oxygen free radicals(OH·)in the three groups were detected by spectrophotometer,immunocytochemical staining,and confocal fluorescence.Western blot and image analysis were used to measure membrane translocation and expression of phospho-protein kinase Ca.MyoFbs incubated with AngII(10^(-8) to 10^(-6) M)for 24 h increased their rate of proliferation,the content of OH·,and expression of ROS(P<0.01 vs.control group),whereas these parameters decreased in the presence of NAC.Immunocytochemistry,confocal fluorescence staining and image analysis showed that AngII could promote the translocation and expression of p-PKCα in membrane,and the antioxidant NAC blocked this increase(P<0.01).Western blot results also showed that NAC could inhibit the expression of p-PKCα. | Liying Wang Hong Li Shijie Yang | 2013 | Acta Pharmaceutica Sinica B2013，3，1： | 0 |
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