| OBJECTIVENod-like receptor protein 3 inflammasome is a multiprotein complex,Which can identify a variety of pathogenic microorganisms and endogenous danger signals,and activate cysteine aspartic acid protease 1(casepase-1),further promote the inflammatory cytokines IL-1 beta and IL-18 release,It plays an important role in innate immunity.The present study found that NLRP3 inflammasome played an important role in a variety of inflammatory related diseases,such as diabetic nephropathy,retinopathy,autoimmune disease,atherosclerosis and reperfusion injury,and so on.The damages of central nervous system included cognitive impairment injury and cerebrovascular disease in Type 2 diabetes.The NLRP3 inflammasome also has studied in the central nervous system diseases.But whether NLRP3 inflammasome works in the damage of brain microvascular system of type 2 diabetes mellitus is not unclear,and the work mechanism is unclear.This experiment aims to discuss the changes and mechanism of NLRP3 inflammasome in the brain microvascular endothelial cells by observing the expression of NLRP3 in the tissues of the rats,cultivating mouse brain microvascular endothelial cells,and testing related proteins in cells.NETHODS(1)A total of 3 months Wistar rats(n=5)and spontaneous type 2diabetes(GK,n=5)rats were enrolled.Using immunohistochemical method to observe the expressions of NLRP3 in the brain tissue.(2)After cerebral microvascular endothelial cells were cultivated in vitro,they were divided into 5 groups:normal control group?high glucose group 1(HG1 group,glucose concentration for 10mmol/L)?high glucose group 2(HG2 group,glucose concentration for 20mmol/L)?high glucose group 3(HG3 group,glucose concentration for 30mmol/L)and HG+NAC group.(3)the expressions of thioredoxin interacting protein(TXNIP)and NLRP3were detected by Western blotting;the level of interleukin-1?(IL-1?)of each group was evaluated by ELISA kits.;intracellular reactive oxygen species(ROS)of normal control group?HG3 group?HG+NAC groupwere detected by flowcytometry.(4)A11 the datas were analyzed by SPSS 19.0 statistical software,and showed by mean±SD.Comparison between two groups is analyzed by t-test,comparison between several groups by one-way analysis of variance and inspection and the correlation is analyzed by Person analysis correlation.All the analysis is with P<0.05 as statistical significance.RESULT(1)NLRP3 mainly distributed in the glial cells of cortex and subcortical tissue and micro/small blood vessel wall,compared with Wistar rat,both the strength and vascular numbers of positive staining were increased(P<0.05)in GK rat.(2)Compared with normal control group,the expressions of TXNIP?NLRP3 increased in HG1?HG2?HG3 group(P<0.01),especially in the HG3 group.(3)Compared with normal control group,the level of IL-1? increased(P<0.01);(4)Compared with normal control group,the level of ROS of HG3 group was increased(P<0.01);Compared with HG3 group,the level of intracellular ROS and IL-1??the expressions of TXNIP?NLRP3 of HG+NAC group significantly decreased(P<0.01).CONCLUSIONS(1)high glucose can promote the expression of NLRP3 and TXNIP,and expressions increased with the increase of the concentration of sugar consentrations.(2)The level of IL-1 beta of each group was increased,demonstrating that NLRP3 inflammasome was activated.(3)High glucose could induce oxidative stress of cerebral microvascular endothelial cells,and product excess ROS.After giving ROS scavenger,the expression of NLRP3?TXNIP and the level of IL-1 beta were decreased,Showing that high glucose could active NLRP3 inflammasome by someway related with ROS.(4)NLRP3 inflammasome was activated in type 2 diabetes mellitus by high glucose-ROS-TXNIP pathway,and promoted the release of inflammatory infactor IL-1?,playing a important role in the damage of brain microvascular. |
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