The Effect of Lycium ruthenicum Murry Anthocyanins on the Apoptosis and Proliferation of H9c2 Cell Induced by Hypoxia

Abstract

Based on the strong antioxidant activity of anthocyanins, this study was aimed to explore the potential influence of Lycium ruthenicum Murry anthocyanins (LRMA) on hypoxia-induced apoptosis and proliferation of H9c2 rat cardiomyocytes. The cell viability was first tested after exposure to normoxia, hypoxia, and hypoxia + LRMA. After being incubated for 12 h under normoxia and 24 h under hypoxia in 50 μg/mL LRMA or 100 μM Salidroside, respectively, LDH content, cell morphology, and cell cycle progression were analyzed. In addition, cellular apoptosis along with the expression of B-cell lymphoma-2 (Bcl2) and BCL2-associated X (Bax) was explored. After that, RNA-seq and bioinformatics analysis revealed the lncRNAs-miRNAs-mRNAs ceRNA network related to hypoxia adaptation of H9c2 cells modulated by LRMA. The final step was to detect the expression of cyclin T2 (CCNT2), cyclin-dependent kinase 9 (CDK9), and forkhead box P1 (FOXP1). The results revealed that LRMA significantly attenuated the reduction of viability and protected H9c2 cells under hypoxic condition. In addition, LRMA not only restored the proliferation activity of H9c2 cells but also promoted apoptosis through stimulating Bcl2 enhancement and Bax inhibition induced by hypoxic stimulation. This study also identified distinct pairs of ceRNA and the hypoxia-related differentially expressed genes regulated by LRMA, including 862 lncRNAs (694 upregulated and 168 downregulated), 7 miRNAs (1 upregulated and 6 downregulated), and 351 mRNAs (226 upregulated and 125 downregulated). The results also suggested that the hypoxia-related genes regulated by LRMA were mainly enriched in cell proliferation and division, cell energy metabolism, inflammatory response, and cell development. However, LRMA could effectively promote the proliferation of H9c2 cells through enhancing CCNT2, CDK9, and FOXP1 under hypoxic conditions. Overall, LRMA could alleviate the proliferation inhibition and apoptosis in hypoxia-induced H9c2 cells.