基于EETs介导的细胞焦亡探讨化瘀祛痰方对ApoE^(-/-)小鼠动脉粥样硬化的影响

Exploration of the effects of Huayu qutan formula on atherosclerosis in ApoE~(-/-) mice based on EET-smediated pyroptosis

目的 探讨化瘀祛痰方对ApoE-/-小鼠动脉粥样硬化(AS)的影响及机制。方法 将30只ApoE-/-小鼠随机分为模型组、化瘀祛痰方组[20 g/(kg·d)]和瑞舒伐他汀组[1.55 mg/(kg·d)],每组10只,另取10只C57BL/6J小鼠作为正常对照组。ApoE-/-小鼠喂饲高脂饲料12周复制AS模型。造模成功后,各组小鼠灌胃相应药物或生理盐水,每日1次,连续4周。末次给药后,检测各组小鼠血清中总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)、5,6-环氧二十碳三烯酸(5,6-EET)、8,9-EET、11,12-EET、14,15-EET、白细胞介素1β(IL-1β)、IL-18水平,主动脉组织中核因子κB抑制蛋白(IκB)、核因子κB(NF-κB)、NOD样受体热蛋白结构域相关蛋白3(NLRP3)、胱天蛋白酶1(Caspase-1)、IL-1β、IL-18 mRNA表达水平和IκB、NF-κB、NLRP3、凋亡相关斑点样蛋白质(ASC)、Caspase-1、消皮素D(GSDMD)、IL-1β、IL-18蛋白表达水平,并观察主动脉组织形态学变化。结果 与模型组比较,化瘀祛痰方组及瑞舒伐他汀组小鼠血清中TC、TG、LDL-C、IL-1β、IL-18水平,主动脉组织中IκB、NF-κB、NLRP3、Caspase-1、IL-1β、IL-18 mRNA表达水平以及IκB、NF-κB、NLRP3、ASC、Caspase-1、GSDMD、IL-1β、IL-18蛋白表达水平均显著降低(P<0.05),血清中5,6-EET、8,9-EET、11,12-EET、14,15-EET水平均显著升高(P<0.05);主动脉粥样斑块明显减少。结论 化瘀祛痰方可能通过EETs介导细胞焦亡发挥抗AS作用。

OBJECTIVE To explore the effects and mechanism of Huayu qutan formula on atherosclerosis(AS) in ApoE-/-mice.METHODS Thirty ApoE-/-mice were randomly divided into model group,Huayu qutan formula group [20 g/(kg·d)],rosuvastatin group [1.55 mg/(kg·d)],with 10 mice in each group.Another 10 C57BL/6J mice were selected as normal control group.ApoE-/-mice were given high-lipid diet for 12 weeks to induce AS model.After modeling,each group was given relevant medicine or normal saline intragastrically,once a day,for 4 consecutive weeks.After the last administration,the levels of total cholesterol(TC),triglycerides(TG),low-density lipoprotein cholesterol(LDL-C),high-density lipoprotein cholesterol(HDLC),5,6-epoxyeicosatrienoic acid(5,6-EET),8,9-EET,11,12-EET,14,15-EET,interleukin-1β(IL-1β) and IL-18 in serum were detected;mRNA expressions of inhibitor of nuclear factor κB(IκB),nuclear factor κB(NF-κB),NOD-like receptor thermal protein domain associated protein 3(NLRP3),Caspase-1,IL-1β and IL-18 in the aortic tissue of mice were detected;protein expression levels of IκB,NF-κB,NLRP3,apoptosis-associated speck-like protein containing CARD(ASC),Caspase-1,gasdermin D(GSDMD),IL-1β and IL-18 in the aortic tissue of mice were detected.The morphological changes of the aortic tissue were observed.RESULTS Compared with model group,the serum levels of TC,TG,LDL-C,IL-1β and IL-18,the m RNA expressions of IκB,NF-κB,NLRP3,Caspase-1,IL-1β and IL-18 in aortic tissue,and the protein expressions of IκB,NF-κB,NLRP3,ASC,Caspase-1,GSDMD,IL-1β and IL-18 were all decreased significantly in Huayu qutan formula group and rosuvastatin group(P<0.05),while the serum levels of 5,6-EET,8,9-EET,11,12-EET and 14,15-EET were increased significantly(P<0.05).The aortic atherosclerotic plaques were alleviated significantly.CONCLUSIONS Huayu qutan formula can play role of anti-AS through EETs-mediated pyroptosis.