Oxymatrine protects H9c2 cardiomyocytes from injury induced by H2O2 by activating Nrf2/HO-1 signaling pathway
ZHANG Zhongbai1,2, LI Yanchun3, CHEN Rundu1, LI Yuanxin1, ZHANG Mei4
1.Logistics Vnivesity of Chinese People’s Armed Police Force, Tianjin 300309, China; 2.Health Team of Detachment of Daxinganling, Daxinganling 165000, China; 3.Department of Pharmacy, Heilongjiang Municipal Corps Hospital of Chinese People’s Armed Police Force, Harbin 150076, China; 4.Department of Cardiology, The Characteristic Medical Center of People’s Armed Police Force, Tianjin 300162, China
Abstract：Objective To investigate the protective effects and mechanisms of oxymatrine (OMT) on H9c2 cardiomyocytes induced by oxidative stress. Methods An oxidative stress model of H9c2 cardiomyocytes was established and pretreated with oxymatrine. Logarithmic growth phase cells were taken and divided into four groups: the control group that was left untreated,model group in which 100 μmol/L H2O2 was added to the cell culture medium for 4 h,and two oxymatrine pretreatment groups that were pretreated for 12 h with complete DMEM medium containing 10 μmol/L and 50 μmol/L oxymatrine respectively before 100 μmol/L H2O2 was added for 4 h. Cell viability was detected by MTT assay while cell morphology was observed by HE staining. The cell supernatant was collected to determine the content of lactate dehydrogenase (LDH) and the levels of malondialdehyde (MDA), catalase (CAT) and superoxide dismutase (SOD) in the cells. The rate of apoptosis was measured by Hoechst. The expression levels of Bcl-2 mRNA, Bax mRNA, Casepase3 mRNA, Nrf2 mRNA and HO-1 mRNA were analyzed by RT-PCR. Protein expressions of Nrf2 and HO-1 were analyzed by Western blot. Results Compared with the control group, the rate of cell viability (39.53%±0.25%) was obviously decreased, the amount of LDH leakage[ (472.22±15.54) U/L]was increased, the content of SOD[ (64.51±5.37) U/mgprot]was decreased and the content of MDA[ (6.70±0.05) nmol/mgprot] was elevated in the model group. The results by RT-PCR and Western blot showed that the expressions of Nrf2 and HO-1 were increased (P<0.05). Compared with the model group, different concentrations in OMT pretreatment groups ameliorated cell viability (57.40%±0.12%,77.51%±0.23%). The amount of LDH leakage [ (367.72±12.53) U/L, (275.35±12.48) U/L] was decreased, the content of SOD[ (90.55±2.27) U/mgprot, (130.52±5.94) U/mgprot] was increased and the content of MDA[ (4.75±0.09) nmol/mgprot, (3.22±0.03) nmol/mgprot] was reduced. The results by RT-PCR and Western blot showed that the expressions of Nrf2 and HO-1 were increased (P<0.05). Conclusions Oxymatrine can protect H9c2 cardiomyocytes induced by oxidative stress by inhibiting mitochondrial apoptosis pathway and activating Nrf2/HO-1 signaling pathway.
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2020, Vol. 31 
