青藤碱通过激活核因子E2相关因子2通路抑制小鼠动脉粥样硬化进展

Sinomenine inhibits atherosclerosis progression in mice by activating nuclear factor E2 related factor 2 pathway

目的:探讨青藤碱对载脂蛋白E基因敲除(ApoE-/-)小鼠动脉粥样硬化形成的影响及其机制。方法:采用随机数字表法将6~8周龄雄性ApoE-/-小鼠分为模型组、青藤碱低剂量组和高剂量组,每组10只。30只小鼠均高脂饲料喂养16周,在高脂饲养8周后,低剂量组和高剂量组分别给予青藤碱50 mg/(kg·d)和100 mg/(kg·d)灌胃,模型组小鼠灌胃等量蒸馏水,持续8周。干预结束后检测各组小鼠血清中血脂水平、氧化应激指标,并采用苏木精-伊红(HE)染色、马松染色及核因子E2相关因子2(Nrf2)免疫组织化学分析斑块形态,蛋白质印迹法(Western blot)检测主动脉Nrf2及下游靶向蛋白的表达。组间比较采用t检验和单因素方差分析。结果:高剂量青藤碱组小鼠体重轻于模型组[(30.910±3.741)g比(37.774±7.182)g,t=0.245,P<0.05],高剂量青藤碱组小鼠血清总胆固醇水平低于模型组[(10.301±2.210)mmol/L比(13.232±2.280)mmol/L,t=1.217,P<0.01]。斑块形态学比较,青藤碱给药组小鼠动脉粥样硬化斑块范围小于模型组,炎性细胞浸润少于模型组,胶原含量多于模型组,主动脉Nrf2及下游靶蛋白表达水平高于模型组。结论:青藤碱可激活Nrf2信号通路,增强抵抗氧化应激的能力,增加斑块稳定性,对动脉粥样硬化具有保护作用。

Objective To investigate the effect of sinomenine on atherosclerosis in apolipoprotein E gene knockout(ApoE-/-)mice and its mechanism.Methods Using a random number table method,male ApoE-/-mice aged 6-8 weeks were divided into model group(n=10),low-dose sinomenine group(n=10)and high-dose sinomenine group(n=10).A total of 30 mice were fed with high-fat diet for 16 weeks.After 8 weeks of high-fat diet,sinomenine[50 mg/(kg·d)and 100 mg/(kg·d)]was given to the low-dose group and high-dose group respectively,and the model group was given the same amount of distilled water for 8 weeks.After the intervention,the serum lipid level and oxidative stress index were detected,and the plaque morphology was analyzed by hematoxylin and eosin(HE)staining,Masson staining and nuclear factor erythroid 2-related factor 2(Nrf2)immunohistochemical staining,and the expression of aortic Nrf2 and downstream targeted protein was detected by Western blotting.T-test and one-way analysis of variance were used for comparison between groups.Results The body weight of mice in high dose sinomenine group was lower than that in model group[(30.910±3.741)g vs.(37.774±7.182)g,t=0.245,P<0.05],and the level of serum total cholesterol in mice of high dose sinomenine group was decreased[(10.301±2.210)mmol/L vs.(13.232±2.280)mmol/L,t=1.217,P<0.01].The sinomenine group showed smaller atherosclerotic plaque,less inflammatory cell infiltration,higher collagen content and expression of Nrf2 and downstream target proteins in aorta.Conclusion Sinomenine can activate Nrf2 signal pathway,enhance the ability to resist oxidative stress,increase plaque stability,and has a protective effect on atherosclerosis.