Nod样受体蛋白3炎性体在2型糖尿病脑微血管内皮细胞中的变化及变化机制

发布时间：2018-04-04 06:15

  本文选题：Nod样受体蛋白炎性体　切入点：型糖尿病　出处：《山东大学学报(医学版)》2017年03期

【摘要】：目的探讨Nod样受体蛋白3(NLRP3)炎性体在2型糖尿病脑微血管内皮中的变化及机制。方法 3月龄Wistar大鼠和自发性2型糖尿病(GK)大鼠各5只,采用免疫组织化学法观察NLRP3在脑组织中的表达;体外培养小鼠脑微血管内皮细胞(CMEC)。不同糖浓度培养基模拟2型糖尿病内环境,活性氧(ROS)阻断剂N-乙酰半胱氨酸(NAC)作为抑制剂,将细胞分为对照组(糖浓度5.6 mmol/L)、高糖1组(HG1组,培养基糖浓度10 mmol/L)、高糖2组(HG2组,培养基糖浓度20 mmol/L)、高糖3组(HG3,培养基糖浓度30 mmol/L)及高糖+NAC组(HG+NAC组,目的蛋白表达较明显组的糖浓度)。采用Western blotting法检测各组硫氧还蛋白交互蛋白(TXNIP)和NLRP3的表达,并对TXNIP和NLRP3是否存在相关性进行分析;采用ELISA法检测白介素1β(IL-1β)的含量;采用流式细胞术检测对照组、HG3组、HG+NAC组ROS的含量。结果 NLRP3在脑微血管壁聚集,与Wistar大鼠相比,GK大鼠的阳染强度,阳染血管数均有增加(P0.05);与对照组相比,HG1组、HG2组、HG3组TXNIP、NLRP3的表达增加(P0.01),HG3组最明显,细胞内IL-1β水平增高(P0.01),ROS的含量增加(P0.01);细胞内TXNIP和NLRP3的表达呈正相关性(r=0.993;P0.05);给予ROS清除剂后,与HG3组相比,HG+NAC组细胞内ROS含量下降(P0.01),TXNIP、NLRP3表达水平下降(P0.01),细胞内IL-1β水平下降(P0.01)。结论 NLRP3在2型糖尿病脑微血管内皮细胞中经ROS-TXNIP途径被激活,并且促进炎性因子IL-β的释放,在2型糖尿病脑微血管损伤中发挥重要作用。
[Abstract]:Objective to investigate the changes and mechanism of Nod like receptor protein 3 (NLRP3) inflammatory body in type 2 diabetic brain microvascular endothelium.Methods the expression of NLRP3 in brain tissue of 3 month old Wistar rats and 5 spontaneous type 2 diabetic rats were observed by immunohistochemical method, and mouse brain microvascular endothelial cells were cultured in vitro.The cells were divided into control group (5. 6 mmol 路L ~ (-1)), HG1 group with high glucose concentration, 10 mmol / L ~ (-1) glucose concentration in medium, 10 mmol / L ~ (-1) glucose concentration in medium and HG2 group in high glucose 2 group.The concentration of glucose was 20 mmol 路L ~ (-1) in culture medium, 30 mmol / L in high glucose group, and 30 mmol 路L ~ (-1) in high glucose group. The protein expression in HG NAC group was higher than that in high glucose NAC group.Western blotting method was used to detect the expression of thioredoxin (TXNIPP) and NLRP3, and the correlation between TXNIP and NLRP3 was analyzed, and the content of IL-1 尾 was detected by ELISA method.The content of ROS in HG NAC group was detected by flow cytometry.缁嗚優鍐匢L-1尾姘村钩澧為珮(P0.01),ROS鐨勫惈閲忓�炲姡�

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