Effect of dihydrobayberry on high glucose-caused injury of H9C2 myocardial cell

Abstract

Objective To investigate the influence of dihydrobayberry (DHM) on high glucose (HG) -induced cardiomyocytes H9c2 cells damage and its poteintial mechanism. Methods Cells were divided into the following groups: control group, 35 mmol/L HG group, 35 mmol/L HG+50 μmol/L DHM group and 50 μmol/L DHM group. CCk-8 assay was used to detect cell viability. The levels of malondialdehyde (MDA) , superoxide dismutase (SOD) and catalase (CAT) were determined by chemical colorimetry. ROS levels were measured by flow cytometry. TNFα, IL1β and IL6 mRNA expression and contents were determined by fluorescence quantitative PCR and Elisa assays respectively. The expression levels of p-IκBα and IκBα proteins, and nucleoprotein NF-κB p65 were detected by Western Blot. Univariate analysis of variance was used for comparison between groups. Results Pretreatment with 50 and 100 μmol/L DHM significantly inhibited the reduced cell viability of H9C2 myocardial cells caused by 35 mmol/L HG: The cell viability of the control group, 35 mmol/L HG group, 35 mmol/L HG+50 μmol/L DHM group, 35 mmol/L HG+100 μmol/L DHM group were (100±0.00) ﹪, (52.23±5.69) ﹪, (74.58±6.12) ﹪ and (86.04±3.76) ﹪, respectively (F = 40.61, P < 0.001) . We also found that pretreatment with DHM (50 μmol/L) significantly inhibited the enhanced MDA and ROS levels, and decreased SOD and CAT activity of H9C2 myocardial cells induced by HG (35 mmol/ L) . The MDA level, SOD and CAT activity of the control group, HG group and HG+ DHM group were (0.44±0.06) nmol/ ml, (2.33±0.40) nmol/ml, (1.48±0.41) nmol/ml, (156.00±9.00) U/ml, (325.3±10.69) U/ ml, (244.0±9.54) U/ml, (10.62±1.59) U/ml, (5.18±0.34) U/ml, (7.75±0.53) U/ ml and (11.31±0.98) U/ ml , (5.20±1.12) U/ml, (8.06±0.66) U/ml, respectively. (F = 30.34, 29.75, 14.72, P all < 0.001) . Pretreatment with DHM significantly inhibited the increased expression levels of TNFα, IL1β and IL6 mRNA and their contents caused by HG in H9C2 myocardial cells (P all < 0.001) . DHM significantly inhibited the increased expression of p-IκBα/IκBα and nucleoprotein NF-κB p65 in H9C2 myocardial cells caused by HG (P all < 0.001) . Conclusion DHM can antagonize HG-induced H9C2 myocardial cell injury, which may be related to its inhibition of NF-κB signaling pathway. Key words: Dihydromyricetin; High glucose; Oxidative stress; Inflammatory response; Myocardial cells