miR-1306-5p靶向水通道蛋白1对糖尿病心肌病变的调控作用

Effects of miR-1306-5p targeting AQP1 in regulating diabetic cardiomyopathy

目的:探究miR-1306-5p对糖尿病心肌病变的影响和可能机制。方法:体外培养人心肌细胞AC16,分为对照组和高糖组,作用24 h后,收集细胞上清液,采用全自动生化分析仪检测心肌酶CK、LDH和AST水平,采用Western blot检测AC16细胞中心肌病变相关蛋白ANF、β-MHC、TGF-β的表达以及水通道蛋白1(AQP1)的表达,采用qRT-PCR检测miR-1306-5p和AQP1 mRNA的表达。采用双荧光素酶实验验证miR-1306-5p与AQP1的靶向关系。另取AC16细胞,观察抑制miR-1306-5p的表达或同时抑制miR-1306-5p和AQP1的表达对心肌酶的分泌、心肌病变相关蛋白表达、凋亡率以及凋亡相关蛋白Bax和Bcl-2表达的影响。结果:高糖处理提高了AC16细胞心肌酶CK、LDH和AST的分泌,心肌病变相关蛋白ANF、β-MHC、TGF-β以及miR-1306-5p的表达,降低了AQP1的表达(P<0.05)。miR-1306-5p与AQP1存在负向靶控关系。抑制miR-1306-5p表达降低了高糖诱导的AC16细胞凋亡率、心肌酶的分泌及心肌病变相关蛋白和Bax蛋白的表达水平,提高了Bcl-2蛋白的表达水平(P<0.05);抑制AQP1逆转了抑制miR-1306-5p对高糖诱导的AC16细胞上述指标的改善作用(P<0.05)。结论:抑制miR-1306-5p表达可能通过靶向上调AQP1减轻高糖诱导的心肌细胞损伤。

Aim:To explore the effects and possible mechanism of miR-1306-5p on diabetic cardiomyopathy.Methods:Myocardial AC16 cells were allocated into control group and high sugar(HG)group.After 24 hours,collecting supernatant,using automatic biochemical analyzer to determine the levels of myocardial enzymes(CK,LDH and AST),using Western blot to detect the expressions of myopathy associated proteins(ANF,β-MHC,and TGF-β),as well as the expression of AQP1 protein,using qRT-PCR to detect the expression of miR-1306-5p and AQP1 mRNA.Dual luciferase assay was used to verify the targeting relationship between miR-1306-5p and AQP1.In addition,AC16 cells were used to observe the effects of inhibition of miR-1306-5p or the simultaneous inhibition of miR-1306-5p and AQP1 on the secretion of myocardial enzymes,the expressions of cardiomyopathy-related proteins,apoptosis rate,and the expressions of apoptotic proteins Bax and Bcl-2.Results:HG treatment increased the secretion of myocardial enzymes,the expressions of cardiomyopathy-related proteins and miR-1306-5p,and decreased the expression of AQP1 in AC16 cells(P<0.05).There was a negative target relationship between miR-1306-5p and AQP1.Inhibition of miR-1306-5p expression decreased the apoptosis rate of AC16 cells induced by HG,the secretion of myocardial enzymes,and the expressions of cardiomyopathy-related proteins and Bax,and increased the expression of Bcl-2(P<0.05).Inhibition of AQP1 reversed the improvement effect of miR-1306-5p on the above indexes in AC16 cells induced by HG(P<0.05).Conclusion:Inhibition of miR-1306-5p expression may reduce myocardial damage induced by HG through targeting and upregulating AQP1.