【作者】 吴娣； 谢凤； 肖凌； 何璐； 刘梅青； 周群； 李兰芳； 陈临溪；

【Author】 Wu Di;Xie Feng;Xiao ling;He Lu;Liu Meiqing;Zhou Qun;Chen Linxi;Institute of Pharmacy and Pharmacology, Learning Key Laboratory for Pharmacoproteomics, University of South China;

【机构】 南华大学药物药理研究所；

【摘要】 Objective: To investigate the inhibition effect and mechanism of caveolin-1 on apelin-13 induced H9 c2 cardiomyocytes hypertrophy and interleukin 8(IL-8) secretion. Methods:We firstly established left ventricular hypertrophy rat model by ligating abdominal aorta, then tested the level of apelin receptor APJ and caveolin-1 in left ventricle of operation and sham rats. Accordingly, we cultured H9 c2 rat cardiomyocytes to detect the expression of caveolin-1 at different concentrations(0, 0.001, 0.01, 0.1, 1μmol/L) and times(0, 3, 6, 12, 24 h) of apelin-13. H9 c2 cells were pre-incubated with caveolae disrupt agent Beta-Cyclodextrins(β-CD) or transfected with pcMV6-Caveolin-1 and then treated with apelin-13(0.1μmol/L, 24 h). These groups were detected by Western blot to examine the expression of caveolin-1, autophagy related proteins Beclin1 and LC3 II/I. Next we applied bioinformatics software STRING to predict the possible interaction between apelin and IL-8. Meanwhile, we tested the effect of apelin-13 on IL-8 secretion by ELISA. Finally, H9 c2 cells were incubated with β-CD, pcMV6-Caveolin-1, autophagy inhibitor(3-MA) before apelin-13 stimulated. The secretion of IL-8 was detected by ELISA and the cardiomyocytes hypertrophy index ANP and BNP were examined by Western blot in these groups. Results:Firstly, we successfully established the left ventricular hypertrophy rat and found that the APJ was increased, yet the caveolin-1 was decreased in cardiac hypertrophy rats. We found that apelin-13 could suppress caveolin-1 expression in a dose and time depend manner in cultured H9 c2 cells as well. Western blot showed that apelin-13 stimulated the expression of Beclin-1 and LC3 II/I, this effect could inhibit by pcMV6-Caveolin-1 transfection. We also observed the apelin-13 stimulated LC3 II/I was further promoted by β-CD. STRING test found that apelin was closely interacted with IL-8, and apelin-13 could significantly promote IL-8 secretion. Additionally, the effects of apelin-13 on IL-8, ANP and BNP were reversed by pcMV6-Caveolin-1 transfection and 3-MA, yet further strengthened by β-CD. Conclusion:Caveolin-1 could depress apelin-13 induced cardiomyocytes hypertrophy and IL-8 secretion, and these effects by suppressing autophagy, which provided a novel treatment target for heart diseases.更多还原

【Abstract】 Objective: To investigate the inhibition effect and mechanism of caveolin-1 on apelin-13 induced H9 c2 cardiomyocytes hypertrophy and interleukin 8(IL-8) secretion. Methods:We firstly established left ventricular hypertrophy rat model by ligating abdominal aorta, then tested the level of apelin receptor APJ and caveolin-1 in left ventricle of operation and sham rats. Accordingly, we cultured H9 c2 rat cardiomyocytes to detect the expression of caveolin-1 at different concentrations(0, 0.001, 0.01, 0.1, 1μmol/L) and times(0, 3, 6, 12, 24 h) of apelin-13. H9 c2 cells were pre-incubated with caveolae disrupt agent Beta-Cyclodextrins(β-CD) or transfected with pcMV6-Caveolin-1 and then treated with apelin-13(0.1μmol/L, 24 h). These groups were detected by Western blot to examine the expression of caveolin-1, autophagy related proteins Beclin1 and LC3 II/I. Next we applied bioinformatics software STRING to predict the possible interaction between apelin and IL-8. Meanwhile, we tested the effect of apelin-13 on IL-8 secretion by ELISA. Finally, H9 c2 cells were incubated with β-CD, pcMV6-Caveolin-1, autophagy inhibitor(3-MA) before apelin-13 stimulated. The secretion of IL-8 was detected by ELISA and the cardiomyocytes hypertrophy index ANP and BNP were examined by Western blot in these groups. Results:Firstly, we successfully established the left ventricular hypertrophy rat and found that the APJ was increased, yet the caveolin-1 was decreased in cardiac hypertrophy rats. We found that apelin-13 could suppress caveolin-1 expression in a dose and time depend manner in cultured H9 c2 cells as well. Western blot showed that apelin-13 stimulated the expression of Beclin-1 and LC3 II/I, this effect could inhibit by pcMV6-Caveolin-1 transfection. We also observed the apelin-13 stimulated LC3 II/I was further promoted by β-CD. STRING test found that apelin was closely interacted with IL-8, and apelin-13 could significantly promote IL-8 secretion. Additionally, the effects of apelin-13 on IL-8, ANP and BNP were reversed by pcMV6-Caveolin-1 transfection and 3-MA, yet further strengthened by β-CD. Conclusion:Caveolin-1 could depress apelin-13 induced cardiomyocytes hypertrophy and IL-8 secretion, and these effects by suppressing autophagy, which provided a novel treatment target for heart diseases.更多还原