清脂化瘀颗粒调控Keap-1/Nrf2/ARE介导的氧化损伤抗动脉粥样硬化的作用及机制研究

Effect and mechanism of Qingzhi Huayu granule on regulating Keap-1/Nrf2/ARE mediated oxidative damage and anti atherosclerosis

目的:基于对动脉粥样硬化(AS)防治,观察清脂化瘀颗粒对动脉粥样硬化的防治作用及机制。方法:将30只载脂蛋白E基因敲除(ApoE^(-/-))小鼠随机分为模型组、阿托伐他汀组、清脂化瘀颗粒组。10只C57BL/6J雄性小鼠作为对照组。除对照组外,各组给予高脂饲料喂养8周复制AS模型,后各组小鼠分别给予对应药物干预4周。分离各组小鼠血清主动脉组织,苏木精-伊红(HE)染色观察主动脉斑块形成情况,比色法检测血清超氧化物歧化酶(SOD)、丙二醛(MDA)、谷胱甘肽(GSH)和亚铁离子(Fe^(2+))水平情况,Western blot检测主动脉组织中核因子E_(2)相关因子(Nrf2)及Kelch样环氧氯丙烷相关蛋白-1(Keap-1)表达情况。结果:与对照组比较,模型组小鼠血清总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白(LDL)水平显著上升,高密度脂蛋白(HDL)水平显著下降,血清SOD、GSH水平显著降低,MDA水平显著升高,Keap-1水平显著升高;主动脉斑块有较大斑块形成。与模型组比较,经过干预后,各组小鼠血清TC、TG、LDL水平显著下降;血清SOD、GSH水平显著升高,MDA水平显著下降,Nrf2蛋白表达显著升高,Keap-1水平显著降低,主动脉斑块减少。与阿托伐他汀组相比,清脂化瘀颗粒组Fe^(2+)水平显著降低。结论:清脂化瘀颗粒组能减轻动脉粥样硬化斑块形成,其机制可能与调控Keap-1/Nrf2/ARE介导的抗氧化损伤机制有关。清脂化瘀颗粒能明显减轻AS的铁蓄积。

Objective:Based on the prevention and treatment of atherosclerosis(AS),to observe the preventive and therapeutic effect and mechanism of Qingzhi Huayu granule on atherosclerosis.Methods:Thirty apolipoprotein E gene knockout(ApoE^(-/-))mice were randomly divided into model group,atorvastatin group and Qingzhi Huayu granule group.Ten C57BL/6J male mice were used as the control group.In addition to the control group,each group was fed with high-fat diet for 8 weeks to replicate the as model,and then the mice in each group were intervened with corresponding drugs for 4 weeks.The serum aortic tissues of mice in each group were separated,the formation of aortic plaque was observed by hematoxylin eosin(HE)staining,the levels of serum superoxide dismutase(SOD),malondialdehyde(MDA),glutathione(GSH)and ferrous ion(Fe^(2+))were detected by colorimetry,and the nuclear factor E_(2) related factor(Nrf2)in aortic tissues was detected by Western blot and kelch like epichlorohydrin associated protein-1(Keap-1).Results:Compared with the control group,the levels of serum total cholesterol(TC),triglyceride(TG)and low density lipoprotein(LDL)in the model group increased significantly,the levels of high density lipoprotein(HDL)decreased significantly,the levels of serum SOD and GSH decreased significantly,the levels of MDA and Keap-1 increased significantly;aortic plaque has large plaque formation.Compared with the model group,the levels of serum TC,TG and LDL-C in each group decreased significantly after intervention.The levels of serum SOD and GSH increased significantly,the level of MDA decreased significantly,the expression of Nrf2 protein increased significantly,the level of keap-1 decreased significantly,and the aortic plaque decreased.Compared with atorvastatin group,Fe^(2+)level in Qingzhi Huayu granule group decreased significantly.Conclusion:Qingzhi Huayu granule group can reduce the formation of atherosclerotic plaque,and its mechanism may be related to the regulation of Keap-1/Nrf2/ARE mediated antioxidant damage mechanism.Qingzhi Huayu granule can significantly reduce the iron accumulation of as.