芍药甘草汤提取物对H_(2)O_(2)诱导心肌细胞氧化应激和炎症反应的影响

Effects of Peony Licorice Decoction Extract on Oxidative Stress and Inflammatory Response of Cardiomyocytes induced by Hydrogen Peroxide

目的:研究芍药甘草汤提取物(SYG)对过氧化氢(hydrogen peroxide,H_(2)O_(2))诱导心肌细胞氧化应激和炎症反应的影响。方法:将心肌细胞分为5组,分别为正常组,模型组(200μmol·L^(-1) H_(2)O_(2))和SYG高、中、低剂量组(150、100、50μg·mL^(-1)),SYG干预24 h后,采用H_(2)O_(2)处理2 h,之后采用MTT法检测细胞增殖能力,紫外分光光度法测定心肌细胞上清液中超氧化物歧化酶(superoxide dismutase,SOD)、过氧化氢酶(catalase,CAT)、乳酸脱氢酶(lactate dehydrogenase,LDH)以及丙二醛(malondialdehyde,MDA)的含量,RT-PCR法测定心肌细胞中肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)、环氧合酶-2(cyclooxygenase-2,COX-2)、核因子-κB(nuclear factor kappa-B,NF-κB)的表达。结果:H_(2)O_(2)诱导可使心肌细胞SOD及CAT含量显著降低,MDA及LDH的含量显著增加,而SYG能够显著改善上述现象,并且能够抑制TNF-α、iNOS、COX-2和NF-κB的表达。结论:SYG能够抑制H_(2)O_(2)诱导后心肌细胞的氧化应激反应及炎症反应,对心肌缺血有一定的保护作用,根据实验结果推测其抗炎作用机制可能为抑制TNF-α激活NF-κB信号通路,从而抑制iNOS及COX-2的表达,但具体作用机制仍需进一步明确。

Objective:To investigate the effect of Peony licorice decoction extract(SYG)on oxidative stress and inflammatory response induced by hydrogen peroxide(H_(2)O_(2))on myocardial cells.Methods:Myocardial cells were divided into five groups,respectively normal group,model group(200μmol·L^(-1) H_(2)O_(2))and SYG groups(150,100,50μg·mL^(-1)),and after the intervention of Peony licorice decoction for 24 h,H_(2)O_(2) was treated for 2 h,then MTT method was used to detect cell proliferation ability,determination of superoxide dismutase(SOD),catalase(CAT),lactate dehydrogenase(LDH)and malondialdehyde(MDA)by UV spectrophotometry.RT-PCR was used to evaluate the inflammatory factor tumor necrosis factor-α(TNF-α),inducible nitric oxide synthase(iNOS),cyclooxygenase-2(COX-2),and nuclear factor kappa-B(NF-κB)in myocardial cells.Results:H_(2)O_(2) induction could significantly reduce the contents of SOD and CAT,and significantly increase the contents of MDA and LDH,while SYG could significantly improve the above phenomena,and SYG could inhibit the expression of TNF-α,iNOS,COX-2 and NF-κB.Conclusion:SYG can inhibit the oxidative stress and inflammatory response of myocardial cells induced by H_(2)O_(2),and has a certain protective effect on myocardial ischemia.It is speculated that its anti-inflammatory mechanism may be related to the inhibition of TNF-αto activate NF-κB pathway to inhibit the expression of iNOS and COX-2,but the specific mechanism still needs to be further clarified.