褪黑素通过激活Akt信号通路缓解高糖诱发的原代心肌细胞损伤

董禹辰; 杨东; 郭建英; 程征

doi:10.12125/j.chj.202005052

褪黑素通过激活Akt信号通路缓解高糖诱发的原代心肌细胞损伤

Melatonin alleviates high glucose-induced primary cardiomyocyte injury via activating the PI3K/p-Akt pathways

摘要

摘要:
目的明确高糖对心肌细胞损伤的影响；揭示褪黑素（Mel）在高糖诱发乳鼠原代心室肌细胞损伤中的作用及机制。
方法体外培养乳鼠原代心室肌细胞，分为4组：正常葡萄糖浓度组(NG)、高糖组(HG，HG=25 mmo/L)、高糖+褪黑素组（HG+Mel，HG=25 mmo/L；Mel=30 μmo/L）、高糖+褪黑素+ PI3K/Akt抑制剂组（HG+Mel，HG=25 mmo/L；Mel=30 μmo/L；LY294002=50 μmo/L）组，采用Western Blotting、RT-PCR和免疫荧光技术检测心肌细胞凋亡相关蛋白以及PI3K/p-Akt等指标，评价褪黑素对高糖诱发心肌细胞凋亡的作用及机制。
结果与NG组比较，HG组心肌细胞Cl -Caspase3、Caspase9的mRNA及蛋白表达明显升高，伴有p-Akt的mRNA及蛋白表达降低和PI3K蛋白表达降低；与HG组相比，HG+Mel组心肌细胞Cl-Caspase3、Caspase9的mRNA及蛋白表达下调伴有p-Akt的mRNA及蛋白表达回升和PI3K蛋白表达升高；与HG+Mel组相比，HG+Mel+LY294002组心肌细胞的Cl-Caspase3、Caspase9的mRNA及蛋白表达上升伴有p-Akt蛋白表达水平显著降低和PI3K蛋白表达降低。免疫荧光的结果与Western Blotting、RT-PCR的结果趋势一致。
结论褪黑素通过激活PI3K/Akt信号通路缓解高糖诱导的原代心肌细胞损伤。

Abstract:
AIM To investigate the effect of high glucose on cardiomyocytes injury and to verify the effects and mechanisms of melatonin regulating high glucose-induced neonatal mouse primary ventricular myocytes injury.
METHODS Neonatal mouse primary ventricular myocytes were cultured in vitro and divided into four groups: normal control group (NG), high glucose group (HG), HG+melatonin group (HG+Mel) and HG+melatonin+PI3k/Akt inhibitor group (HG+Mel+LY294002). Western blotting, RT-PCR and immunofluorescence were used to detect the expressions of apoptosis-associated proteins and the change of PI3K/p-Akt, evaluating the effects and mechanisms of melatonin on high glucose-induced cadiomyocytes apoptosis.
RESULTS Compared with NG, the mRNA/protein expressions ofCl-Caspase3 and Caspase9 increased in cardiomyocytes of high glucose group, accompanied with down-regulated mRNA/protein expressions of p-Akt and decreased protein expression of PI3K. Compared with high glucose group, the mRNA/protein expressions of CL-Caspase3 and Caspase9 in cardiomyocytes of HG+Mel group decreased, together with elevated mRNA/protein expressions of p-Akt and increased protein expression of PI3K. Compared with HG+Mel group, the mRNA/protein expressions of CL-Caspase3 and Caspase9 in cardiomyocytes of HG+Mel+LY294002 group increased, together with the decreased mRNA/protein expressions of p-Akt and down-regulated protein expression of PI3K. The results of immunofluorescence were consistent with Western blotting and RT-PCR.
CONCLUSION Melatonin alleviates high glucose-induced primary cardiomyocytes injury via activating the PI3K/Akt pathway.

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