Oxymatrine protects H9c2 cardiomyocytes from injury induced by H2O2 by activating Nrf2/HO-1 signaling pathway

ZHANG Zhongbai; LI Yanchun; CHEN Rundu; LI Yuanxin; ZHANG Mei

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Medical Journal of the Chinese People Armed Police Forces ›› 2020, Vol. 31 ›› Issue (2) : 139-144.

ORIGINAL ARTICLES

Oxymatrine protects H9c2 cardiomyocytes from injury induced by H₂O₂ by activating Nrf2/HO-1 signaling pathway

ZHANG Zhongbai^1,2, LI Yanchun³, CHEN Rundu¹, LI Yuanxin¹, ZHANG Mei⁴

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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.

Key words

oxymatrine / oxidative stress / cardiomyocyte / protection

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ZHANG Zhongbai, LI Yanchun, CHEN Rundu, LI Yuanxin, ZHANG Mei. Oxymatrine protects H9c2 cardiomyocytes from injury induced by H₂O₂ by activating Nrf2/HO-1 signaling pathway[J]. Medical Journal of the Chinese People Armed Police Forces. 2020, 31(2): 139-144

References

[1] Mendis S, Puska P, Norrving B. Global atlas on cardiovascular disease prevention and control[M]. Geneva: The world health organization, 2011: 4.
[2] Cesselli D, Jakoniuk I, Barlucchi L, et al. Oxidative stress-mediated cardiac cell death is a major determinant of ventricular dysfunction and failure in dog dilated cardiomyopathy[J]. Circ Res, 2001, 89(3): 279-286.
[3] Ma A, Yang Y, Wang Q, et al. Anti-inflammatory effects of oxymatrine on rheumatoid arthritis in rats via regulating the imbalance between Treg and Th17 cells[J]. Mol Med Rep, 2017, 15(6): 3615-3622.
[4] Li W, Yu X, Tan S, et al. Oxymatrine inhibits non-small cell lung cancer via suppression of EGFR signaling pathway[J]. Cancer Med, 2018, 7(1): 208-218.
[5] Wang H, Li Y, Dun L, et al. Antinociceptive effects of oxymatrine from sophora flavescens, through regulation of NR2B-containing NMDA receptor-ERK/CREB signaling in a mice model of neuropathic pain[J]. Phytomedicine, 2013, 20(11): 1039-1045.
[6] Huang X Y, Chen C X. Effect of oxymatrine, the active component from radix sophorae flavescentis (Kushen), on ventricular remodeling in spontaneously hypertensive rats[J]. Phytomedicine, 2013, 20(3-4): 202-212.
[7] Runtao G, Guo D, Jiang B Y, et al. Oxymatrine, the main alkaloid component of sophora roots, protects heart against arrhythmias in rats[J]. Planta Med, 2011, 77(3): 226-230.
[8] Jiang Y, Zhu Y, Mu Q, et al. Oxymatrine provides protection against Coxsackievirus B3-induced myocarditis in BALB/c mice[J]. Antiviral Res, 2017, 141: 133-139.
[9] 张鸣号,王秀玉,姜怡邓,等. JAK/STAT信号通路活化在感染性休克大鼠心肌损伤中的作用及氧化苦参碱干预的实验研究[J]. 中国病理生理杂志,2012,28(11): 2080.
[10] Zhou R, Xu Q, Xu Y, et al. Oxymatrine attenuated isoproterenol-induced heart failure in rats via regulation of COX-2/PGI2 pathway[J]. Biomed Pharmacother, 2016, 84: 1359-1366.
[11] 赵阳. 氧化苦参碱对H9c2心肌细胞氧化应激损伤的保护作用及其机制[J].山东医药, 2015, 55(37): 29-31.
[12] Lee J H, Khor T O, Shu L, et al. Dietary phytochemicals and cancer prevention: Nrf2 signaling, epigenetics, and cell death mechanisms in blocking cancer initiation and progression[J]. Pharmacol Ther, 2013, 137(2): 153-171.
[13] Xu J, Tian W, Ma X, et al. The molecular mechanism underlying morphine-induced Akt activation: roles of protein phosphatases and reactive oxygen species[J]. Cell Biochem Biophys, 2011, 61(2): 303-311.
[14] Stern M M, Myers R L, Hammam N, et al. The influence of extracellular matrix derived from skeletal muscle tissue on the proliferation and differentiation of myogenic progenitor cells ex vivo[J]. Biomaterials, 2009, 30(12): 2393-2399.
[15] Bonavita F, Stefanelli C, Giordano M, et al.H9c2 cardiac myoblasts undergo apoptosis in a model of ischemia consisting of serum deprivation and hypoxia: inhibition by PMA[J]. FEBS Lett, 2003, 536(1-3):85-91.
[16] Javadov S, Hunter J C, Barreto T G, et al. Targeting the mitochondrial permeability transition: cardiac ischemia-reperfusion versus carcinogenesis[J]. Cell Physiol Biochem, 2011, 27(3-4): 179-190.
[17] Liao Y H, Xia N, Zhou S F, et al. Interleukin-17A contributes to myocardial ischemia/reperfusion injury by regulating cardiomyocyte apoptosis and neutrophil infiltration[J]. J Am Coll Cardiol, 2012, 59(4): 420-429.
[18] Li T, Chen L, Yu Y, et al. Resveratrol alleviates hypoxia/reoxygenation injury-induced mitochondrial oxidative stress in cardiomyocytes[J]. Mol Med Rep, 2019, 19(4): 2774-2780.
[19] Dai D F, Chiao Y A, Marcinek D J, et al. Mitochondrial oxidative stress in aging and healthspan[J]. Longev Healthspan, 2014, 3: 6.
[20] Kansanen E, Kuosmanen S M, Leinonen H, et al. The Keap1-Nrf2 pathway: mechanisms of activation and dysregulation in cancer[J]. Redox Biol, 2013, 1: 45-49.
[21] Fan J,Xu G,Jiang T,et al.Pharmacologic induction of heme oxygenase-1 plays a protective role in diabetic retinopathy in rats[J]. Invest Ophthalmol Vis Sci, 2012,53(10): 6541-6556.