本文選題：Nod樣受體蛋白炎性體　切入點(diǎn)：型糖尿病　出處：《山東大學(xué)學(xué)報(bào)(醫(yī)學(xué)版)》2017年03期

【摘要】：目的探討Nod樣受體蛋白3(NLRP3)炎性體在2型糖尿病腦微血管內(nèi)皮中的變化及機(jī)制。方法 3月齡Wistar大鼠和自發(fā)性2型糖尿病(GK)大鼠各5只,采用免疫組織化學(xué)法觀察NLRP3在腦組織中的表達(dá);體外培養(yǎng)小鼠腦微血管內(nèi)皮細(xì)胞(CMEC)。不同糖濃度培養(yǎng)基模擬2型糖尿病內(nèi)環(huán)境,活性氧(ROS)阻斷劑N-乙酰半胱氨酸(NAC)作為抑制劑,將細(xì)胞分為對(duì)照組(糖濃度5.6 mmol/L)、高糖1組(HG1組,培養(yǎng)基糖濃度10 mmol/L)、高糖2組(HG2組,培養(yǎng)基糖濃度20 mmol/L)、高糖3組(HG3,培養(yǎng)基糖濃度30 mmol/L)及高糖+NAC組(HG+NAC組,目的蛋白表達(dá)較明顯組的糖濃度)。采用Western blotting法檢測(cè)各組硫氧還蛋白交互蛋白(TXNIP)和NLRP3的表達(dá),并對(duì)TXNIP和NLRP3是否存在相關(guān)性進(jìn)行分析;采用ELISA法檢測(cè)白介素1β(IL-1β)的含量;采用流式細(xì)胞術(shù)檢測(cè)對(duì)照組、HG3組、HG+NAC組ROS的含量。結(jié)果 NLRP3在腦微血管壁聚集,與Wistar大鼠相比,GK大鼠的陽(yáng)染強(qiáng)度,陽(yáng)染血管數(shù)均有增加(P0.05);與對(duì)照組相比,HG1組、HG2組、HG3組TXNIP、NLRP3的表達(dá)增加(P0.01),HG3組最明顯,細(xì)胞內(nèi)IL-1β水平增高(P0.01),ROS的含量增加(P0.01);細(xì)胞內(nèi)TXNIP和NLRP3的表達(dá)呈正相關(guān)性(r=0.993;P0.05);給予ROS清除劑后,與HG3組相比,HG+NAC組細(xì)胞內(nèi)ROS含量下降(P0.01),TXNIP、NLRP3表達(dá)水平下降(P0.01),細(xì)胞內(nèi)IL-1β水平下降(P0.01)。結(jié)論 NLRP3在2型糖尿病腦微血管內(nèi)皮細(xì)胞中經(jīng)ROS-TXNIP途徑被激活,并且促進(jìn)炎性因子IL-β的釋放,在2型糖尿病腦微血管損傷中發(fā)揮重要作用。
[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.緇嗚優(yōu)鍐匢L-1尾姘村鉤澧為珮(P0.01),ROS鐨勫惈閲忓鍔，

本文編號(hào)：1708724