NLRP3炎症小体对2型糖尿病动脉粥样硬化的影响及作用机制研究

The Effect and Mechanism of NLRP3 Inflammasome on Atherosclerosis in Type 2 Diabetes Mellitus

目的 探究NLRP3炎症小体对2型糖尿病(T2DM)动脉粥样硬化(AS)的影响及其具体作用机制。方法 研究共纳入住院或门诊患者60例,其中T2DM患者30例,T2DM+AS患者30例,同时纳入健康志愿者20名为健康对照组。比较两组患者空腹血糖、糖化血红蛋白、甘油三酯、总胆固醇、低/高密度脂蛋白,彩超检测颈动脉血管壁内中膜厚度,ELISA试剂盒检测血浆NLRP3炎症小体水平。将36只GK大鼠随机分为模型组、NLRP3过表达组、IL-1β抑制剂组,每组12只,均给予高脂饲料诱导早期动脉粥样硬化。12只Wistar大鼠作为空白对照组,正常饲养。检测大鼠空腹血糖、甘油三酯、总胆固醇、低/高密度脂蛋白的含量,ELISA检测IL-1β、NLRP3、IL-18水平,观察大鼠腹主动脉病理改变,Western blot法检测大鼠腹主动脉NLRP3、Caspase3蛋白表达情况。结果 与健康对照组相比,T2DM组和T2DM+AS组患者空腹血糖、糖化血红蛋白、甘油三酯、总胆固醇、低密度脂蛋白含量升高,高密度脂蛋白含量降低,颈动脉血管壁内中膜厚度增加、NLRP3炎症小体水平升高(P <0.05);与T2DM组相比,T2DM+AS组甘油三酯、总胆固醇含量明显增加,颈动脉血管壁内中膜厚度显著增加,NLRP3炎症小体水平升高(P <0.05),其他指标无显著变化。与模型组大鼠比较,NLRP3过表达组大鼠空腹血糖、甘油三酯、总胆固醇、低密度脂蛋白的含量均上升,高密度脂蛋白的含量下降,IL-1β、NLRP3、IL-18水平升高,大鼠腹主动脉病理损伤加重,Caspase1、IL-1β、IL-18蛋白表达上调;与模型组大鼠比较,IL-1β抑制剂组大鼠空腹血糖、甘油三酯、总胆固醇、低密度脂蛋白的含量均下降,高密度脂蛋白的含量上升,IL-1β、IL-18水平降低,大鼠腹主动脉病理损伤缓解,IL-1β、IL-18蛋白表达下调(P <0.05)。结论 NLRP3炎症小体参与调控了2型糖尿病患者动脉粥样硬化的损伤过程,其作用机制可能与Caspase-1/IL-1β/IL-18信号通路有关。

Objective To explore the NLRP3 inflammatory corpuscle effect on atherosclerosis in type 2 diabetes and its specific mechanism of action.Methods A total of 60 inpatients or outpatients were enrolled,including 30 patients with T2DM,30 patients with T2DM+AS,and 20 healthy volunteers as healthy control group.Compare two groups of patients with fasting glucose,glycosylated hemoglobin,triglyceride,cholesterol,low/high density lipoprotein,colour to exceed detect carotid artery wall intima-media thickness,ELISA kit NLRP3 inflammatory corpuscle test.Thirtysix GK rats were randomly divided into model group,NLRP3 overexpression group and IL-1β inhibitor group,with 12 rats in each group.All rats were given high-fat diet to induce early atherosclerosis.Twelve Wistar rats served as the blank control group and were fed normally.Detect rats fasting blood sugar,triglyceride,cholesterol,low/high density lipoprotein content,ELISA detection IL-1β,NLRP3,IL-18 levels,observed the pathological change,big abdomen aorta Western blot method to detect large abdomen aorta NLRP3,Caspase3 protein expression.Results Compared with the healthy control group,the levels of fasting blood glucose,glycosylated hemoglobin,triglyceride,cholesterol and low-density lipoprotein were increased,the levels of high-density lipoprotein were decreased,the intima-media thickness of carotid artery wall was increased,and the level of NLRP3 inflammasome was increased in the T2DM group and T2DM+AS group(P<0.05).Compared with the T2DM group,the T2DM+AS group had significant increases in the contents of triglyceride and cholesterol,intima-media thickness of carotid artery wall,and NLRP3inflammasome level(P<0.05),and no significant changes in other indicators were found.Compared with model group rats,NLRP3 express group rats fasting blood sugar,triglyceride,cholesterol,low density lipoprotein(LDL) levels were significantly increased,the content of HDL decreased significantly,a significant rise in IL-1β,NLRP3,IL-18 levels,big abdomen aorta pathological damage is aggravating,Caspase1,IL-1β,IL-18 protein expression significantly increase,IL-1β inhibitor group rats fasting blood sugar,triglyceride,cholesterol,low density lipoprotein(LDL) levels were significantly decline,a marked increase in the content of HDL,IL-1β,IL-18 levels decreased significantly,big abdomen aorta pathological damage mitigation,the expression of IL-1β and IL-18 protein was significantly down-regulated(P<0.05).Conclusions NLRP3inflammasome is involved in the regulation of atherosclerosis in patients with type 2 diabetes mellitus,and its mechanism may be related to Caspase-1/IL-1β/IL-18 signaling pathway.