论文部分内容阅读
目的:研究糖尿病大鼠心肌细胞小窝蛋白(Caveolin-3,Cav-3)与蛋白激酶C(PKC)β2蛋白的变化并探寻两者的相互作用关系。方法:20只SD雄性大鼠[(250±10)g]随机分为正常对照组(Control)与糖尿病组(Diabetes),分离纯化心肌细胞。Western blot检测Cav-3,PKCβ2蛋白表达水平;免疫共沉淀、免疫荧光与共聚焦观察及Caveolins成分蛋白分析Cav-3与PKCβ2的相互作用关系。结果:与Control组比较,Diabetes组大鼠心肌细胞pPKCβ2表达水平增加而Cav-3表达水平降低。免疫共沉淀结果显示糖尿病促进PKCβ2与Cav-3形成蛋白复合体,免疫荧光与共聚焦观察发现高糖促使PKCβ2向Cav-3蛋白转位增加,进一步Caveolins成分蛋白分析发现PKCβ2共表达于Cav-3蛋白。结论:糖尿病心肌细胞PKCβ2活化水平增加及Cav-3表达水平降低,Cav-3可能参与了糖尿病心肌PKCβ2活化信号向下游分子的的转导。
Objective: To investigate the changes of Caveolin-3 (Cav-3) and protein kinase C (PKC) β2 in diabetic rats and to explore their interaction. Methods: Twenty male Sprague-Dawley rats (250 ± 10 g) were randomly divided into normal control group and Diabetes group. Cardiomyocytes were isolated and purified. Western blot was used to detect the expression of Cav-3 and PKCβ2, and co-immunoprecipitation, immunofluorescence and confocal microscopy were used to detect the interaction between Cav-3 and PKCβ2. Results: Compared with Control group, the expression of pPKCβ2 in Cav-3 and the expression of Cav-3 in Diabetes group decreased. Immunoprecipitation results showed that diabetes mellitus promoted the formation of protein complexes of PKCβ2 and Cav-3. The results of immunofluorescence and confocal analysis showed that high glucose promoted translocation of PKCβ2 to Cav-3 protein. Further analysis of Caveolins protein showed that PKCβ2 was co-expressed in Cav-3 protein . CONCLUSION: The increase of PKCβ2 activation and the decrease of Cav-3 expression in diabetic cardiomyocytes may play a role in the transduction of PKCβ2 activation signal to downstream molecules in diabetic myocardium.