动脉粥样硬化小鼠主动脉平滑肌上瞬时受体电位通道-1-大电导钙离子激活钾通道信号复合体分子表达量的变化研究

Expression of TRPC1-BK Signal Complex in Aortic Smooth Muscle Tissue in Experimental Mice of Atherosclerosis

目的:观察动脉粥样硬化(AS)小鼠主动脉平滑肌上瞬时受体电位通道-1-大电导钙离子激活钾通道(TRPC1-BK)信号复合体分子表达量的变化。方法:AS组采用载脂蛋白E基因缺陷(ApoE^(-/-))小鼠,对照组采用野生型C57BL/6J小鼠,每组10只,分别采用高脂饲料和普通饲料喂养10周,处死后分离主动脉血管平滑肌组织,提取总核糖核酸(RNA)及总蛋白。使用反转录-聚合酶链式反应(RT-PCR)、蛋白免疫印迹(Western blot)及免疫组化染色等方法检测并比较两组TRPC1、大电导钙离子激活钾通道α亚单位(BKα)及大电导钙离子激活钾通道β_1亚单位(BKβ_1)亚基信使核糖核酸(mRNA)和蛋白表达量的差异。结果:AS组AS模型建立成功。RT-PCR检测结果:与对照组比,AS组TRPC1的mRNA表达量增多(P<0.05),BKα和BKβ_1亚基的mRNA表达量减少(P均<0.01)。Western blot法检测结果:与对照组比,AS组TRPC1蛋白表达量增多(P<0.05),BKα和BKβ_1蛋白表达量减少(P均<0.01)。免疫组化染色检测结果:TRPC1蛋白表达量增多(P<0.01),BKα蛋白表达量减少(P<0.01),BKβ_1蛋白表达量减少(P<0.05)。结论:AS组小鼠主动脉平滑肌组织中TRPC1-BK信号复合体的mRNA和蛋白表达量均受影响。推测TRPC1-BK信号复合体有可能成为治疗AS相关性血管疾病的一个新靶点。

Objective： To observe the expression of transient receptor potential charmel-1-large conductance Ca^2＋-activated K^＋ channel （TRPC1-BK） signal complex in aortic smooth muscle tissue in experimental mice of atherosclerosis （AS）. Methods： Our research included in 2 groups： Control group, wild C57BL/6J mice were fed by normal diet and AS group, ApoE^-/- mice were fed by high fat diet to establish AS model. All animals were treated for 10 weeks, n=10 in each group. The total RNA and protein were extracted from aortic smooth muscle tissue in sacrificed mice. The mRNA and protein expressions of TRPC1, BKu and BKβ1 subunit were measured by RT-PCR, Western blot analysis and immunohistochemistry staining. Results： By RT-PCR examination, compared with Control group, AS group showed increased mRNA expression of TRPC1, P〈0.05 and decreased mRNA expressions of BKα and BKβ1 subunit, both P〈0.01. By Western blot analysis, compared with Control group, AS group had elevated protein expression of TRPC 1, P〈0.05 and reduced protein expressions of BKα and BKβ1 subunit, both P〈0.01; by immunohistochemistry staining, AS group had the higher protein expression of TRPC 1, P〈0.01. and the lower protein expressions of BKα and BKβ1 subunit, both P〈0.05. Conclusion： The mRNA and protein expressions of TRPC1-BK signal complex were affected in aortic smooth muscle tissue in AS mice, which speculated that TRPC1-BK signal complex might be become a new target for treating the relevant vascular disease.