去肾交感神经对2型糖尿病大鼠血管内皮细胞自噬及NLRP3活化的影响  

Effects of renal denervation on the vascular endothelial cell autophagy and NLRP3 activation in type 2 diabetic rats

作　　者：王勇[1] 牛伟华 卢成志[2] 许梦萍 徐建强[2] 何强[2] 许学升[1] WANG Yong;NIU Weihua;LU Chengzhi;XU Mengping;XU Jianqiang;HE Qiang;XU Xuesheng(Department of Cardiology,the First People's Hospital of Shangqiu,Shangqiu 476100,China;Department of Cardiology,Tianjin First Central Hospital)

机构地区：[1]商丘市第一人民医院心血管内科,476100 [2]天津市第一中心医院心血管内科

出　　处：《天津医药》2022年第8期810-816,共7页Tianjin Medical Journal

基　　金：国家自然科学基金资助项目(81970303)。

摘　　要：目的探讨去肾交感神经(RDN)对2型糖尿病(T2DM)大鼠血管内皮细胞自噬及核苷酸结合寡聚化结构域受体蛋白3(NLRP3)炎症小体活化的影响。方法采用高脂饮食联合小剂量链脲佐菌素(STZ)腹腔注射制备T2DM大鼠模型。采用随机数字表法分为对照组(CON组)、糖尿病对照组(T2DM组)、双侧假手术组(Sham组)、双侧手术组(RDN组),每组6只。分别于基线状态、RDN术前及术后4周测量各项指标的变化。采用离体血管环张力实验评价血管舒张功能,Western blot检测主动脉内皮细胞自噬蛋白Beclin1、微管相关蛋白1轻链3(LC3)、p62、炎症小体NLRP3、胱天蛋白酶1(Caspase-1),白细胞介素(IL)-1β及内皮型一氧化氮合酶(eNOS)蛋白表达水平。结果与CON组比较,T2DM组大鼠主动脉内皮舒张功能明显降低,自噬蛋白Beclin1、LC3表达降低,p62表达水平升高,且NLRP3、Caspase-1、IL-1β蛋白表达水平升高,eNOS蛋白表达水平降低(P<0.05)。与T2DM组和Sham组相比,RDN组血管内皮舒张功能改善(P<0.05);自噬蛋白Beclin1、LC3表达升高,p62表达水平降低,且NLRP3、Caspase-1、IL-1β蛋白的表达降低,eNOS蛋白表达水平升高(P<0.05)。结论T2DM大鼠血管内皮功能异常可能与自噬水平降低及NLRP3炎症小体的过度活化有关。RDN可以增强血管内皮细胞自噬水平,抑制NLRP3炎症小体的活化,改善血管内皮功能。Objective To explore the effects of renal denervation(RDN)on the autophagy of vascular endothelial cells and NLRP3 inflammatory complexes in type 2 diabetic mellitus(T2DM)rats.Methods T2DM rat model was established by high-fat diet(HFD)combined with low-dose streptozotocin(STZ)intraperitoneal injection.Rats were randomly divided into four groups:the control group(CON,n=6),the diabetic group(T2DM,n=6),the bilateral sham surgery group(Sham,n=6)and the bilateral RDN group(RDN,n=6).Changes in all indicators were recorded at baseline,before and after RDN and 4 weeks after RDN,respectively.In vitro vascular ring tension test was used to evaluate vascular relaxation function.Western blot assay was used to detect the aorta endothelial cell autophagy of Beclin1,LC3 and p62,and the expression of NLRP3 protein and cysteine aspartate protease 1(Caspase-1)and endothelial nitric oxide synthase(eNOS)protein.Results Compared with the control group,the endothelium diastolic function of the thoracic aorta was significantly decreased in the T2DM group,the expression of autophagy-related proteins Beclin1 and LC3 were decreased,the expression of p62 was increased.The expression levels of NLRP3,Caspase-1 and IL-1βprotein were higher in the T2DM group than those of the control group(P<0.05),the level of eNOS protein was decreased(P<0.05).Compared with the T2DM group and the Sham group,the endothelium diastolic function was significantly improved in the RDN group,and the expression levels of Beclin1 and LC3 were increased,the expression of p62 was decreased.The protein expression levels of NLRP3,Caspase-1 and IL-1βwere decreased,the level of eNOS increased(P<0.05).Conclusion The vascular endothelial dysfunction in T2DM ratsmay be associated with the decreased autophagy level and overexpression of NLRP3 inflammatory complex protein.RDN can enhance the autophagy in vascular endothelial cells,decrease the expression of NLRP3 inflammatory complex protein and improve vascular endothelial dysfunction.

关 键 词：糖尿病 2型 血管舒张 内皮细胞 NLR家族 热蛋白结构域包含蛋白3 自噬 去肾交感神经 

分 类 号：R543[医药卫生—心血管疾病]