JOURNAL ARTICLE

Tetramethylpyrazine partially relieves hypoxia-caused damage of cardiomyocytes H9c2 by downregulation of miR-449a

Xuesong Zhang, Huawei Dong, Yong Liu, Junxia Han, Shouyi Tang, Jingna Si
Journal of Cellular Physiology 2019 February 15
30770559
Inadequate oxygen supply is probably one of the most important pathophysiological mechanisms of cardiomyocyte damage in ischemic heart disease. Tetramethylpyrazine (TMP, also known as ligustrazine) is the main active ingredient isolated from the rhizome of Ligusticum chuanxiong Hort. A previous study reported that the TMP could exert cardioprotective activity. This study aimed to explore the molecular mechanism of the protective effects of TMP on cardiomyocyte damage caused by hypoxia. The viability and apoptosis of cardiomyocytes H9c2 were detected using cell counting kit-8 assay and annexin V-FITC/PI staining, respectively. Quantitative reverse transcription polymerase chain reaction was conducted to measure the expression level of microRNA-449a (miR-449a). Cell transfection was performed to upregulate the expression level of miR-449a or downregulate the expression level of sirtuin 1 (Sirt1). The protein expression levels of Sirt1 and key factors involved in cell apoptosis and phosphatidylinositol 3-kinase/protein kinase 3 (PI3K/AKT) pathway were evaluated using western blot analysis. We found that the hypoxia incubation inhibited H9c2 viability, induced cell apoptosis, and inactivated the PI3K/AKT pathway. TMP treatment partially relieved the hypoxia-caused H9c2 cell viability loss and apoptosis, as well as reversed the hypoxia-caused inactivation of the PI3K/AKT pathway. Moreover, TMP partially alleviated the upregulation of miR-449a in H9c2 cells caused by hypoxia. Overexpression of miR-449a weakened the effects of TMP on hypoxia-treated H9c2 cells. Furthermore, Sirt1 was a target gene of miR-449a. Knockdown of Sirt1 also weakened the effects of TMP on hypoxia-treated H9c2 cells. In conclusion, TMP partially relieved hypoxia-caused cardiomyocytes H9c2 viability loss and apoptosis at least through downregulating miR-499a, upregulating Sirt1, and then activating the PI3K/AKT pathway.

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