摘要
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。
Objective： To discuss the effect of taurine（Tau） on the proliferation of hypoxia-induced pulmonary artery smooth muscle cells（PASMCs）,and study whether the extracellular signal-regulated kinase 1 /2（ERK1 /2） signal pathway participated in the Tau-inhibited PASMC proliferation process and the possible molecular mechanism.Method： The primary culture was performed for PASMCs in rats.The second to fifth generations were adopted for the experiment.The Tau concentration was 80 mmol·L- 1.The concentration of ERK1 /2 blocker（ PD98059） was 50 μmol · L- 1.The drug administration time was 24 h.The effect of Tau on the PASMC proliferation was detected by MTT assay,immunofluorescence staining method and western blot under different conditions.The PASMCs were growing were divided into four groups： the normoxia group,the normoxia ＋ Tau group,the hypoxia group and the hypoxia ＋ Tau group.The Western blot was adopted to detect whether the ERK1 /2 signal pathway participated in the Tau-inhibited PASMC proliferation process.Subsequently,the PASMCs were divided into five groups： the normoxia group,the hypoxia group,the hypoxia ＋ Tau group,the hypoxia ＋ Tau ＋ PD98059 group and the hypoxia ＋ PD98059 group.Result： Hypoxia could induce the PASMC proliferation.Under the conditions of normoxia,Tau had no effect on the PASMC proliferation.Under the conditions of normoxia and hypoxia,Tau had no effect on the expression of the tumor necrosis factor-α（TNF-α） among PASMCs.Tau could reverse the expression up-regulation of hypoxia-induced proliferative cell nuclear antigen（ PCNA）（ P 0.01） and Cyclin A（ Cyclin A）（ P 0.05）.Under the conditions of normoxia,Tau had no effect on the expression of phosphoryl extracellular signal-regulated kinase 1 /2（p-ERK1/2）.Hypoxia could up-regulate the p-ERK1/2 expression（P 0.01）.Tau could reverse the up-regulation of the hypoxiainduced p-ERK1 /2 expression（P 0.01）.Both PD98059 and Tau could inhibit the up-regulated expressions of PCNA,Cyclin A and p-ERK1 /2.According to the comparison between the single addition of Tau and PD98059 under conditions of hypoxia,the hypoxia ＋Tau ＋ PD98059 group showed more significant down-regulation in the expressions of PCNA,Cyclin A and p-ERK1 /2.Conclusion：Tau could inhibit the hypoxia-induced PASMC proliferation,and may regulate it through ERK1 /2 pathway.
出处
《中国中药杂志》
CAS
CSCD
北大核心
2014年第10期1902-1907,共6页
China Journal of Chinese Materia Medica
基金
国家自然科学基金项目(81270113
91339107)
研究生创新基金项目(YJSCX2012-ZZ8HSD)
哈尔滨科技创新人才项目(2013RFXXJ092)
大庆重点项目(SCXZ01011)
