摘要
目的探讨EZH2介导NLRP3炎症小体在高糖(HG)诱导大鼠心肌细胞(H9c2)损伤中的作用及其机制。方法CCK8法建立HG诱导H9c2细胞损伤模型,将细胞分为对照(CON)组、HG组,分别用5.5 mmol/L、33 mmol/L葡萄糖干预,再将33 mmol/L葡萄糖干预的H9c2细胞按照脂质体转染法分别转染si-NC、si-EZH2、NLRP3-NC和NLRP3-OE,记为A组、B组、C组、D组、E组、F组。检测细胞凋亡情况、EZH2、NLRP3炎症小体、Cleaved caspase-3、Bax蛋白表达、IL-1β和IL-18 mRNA表达。结果EZH2、NLRP3、ASC、caspase-1及IL-1β、IL-18 mRNA表达比较,CON组低于HG组(t=6.359、2.941、3.795、4.539、17.874、22.507,P<0.05)。B组低于HG组(t=3.168、6.829、3.639、4.737、12.348、13.915、11.378,P<0.05),B组低于A组(t=3.727、5.336、8.485、4.859、11.300、9.831,P<0.05),D组高于C组(t=19.163、10.257、5.743、9.304、16.382、21.493,P<0.05);Cleaved caspase-3、Bax蛋白表达及凋亡率比较,HG组>A组>B组>CON组,HG组与B组无统计学意义(P>0.05),其余均有统计学意义(P<0.05)。与E组比较,F组凋亡率、EZH2、NLRP3、ASC、caspase-1、Cleaved caspase-3、Bax蛋白表达及IL-1β、IL-18 mRNA表达均升高(t=11.658、9.212、6.333、10.592、11.457、17.422、13.163、22.763、9.163,P<0.05)。结论在高糖环境下,EZH2蛋白表达升高,沉默EZH2可通过调节表观遗传修饰靶向抑制NLRP3炎症小体介导的H9c2心肌细胞凋亡和炎症反应损伤。
Objective To investigate the role and mechanism of NLRP3 inflammatory body mediated by EZH2 in the injury of rat cardiomyocytes(H9c2)induced by high glucose(HG).Methods The model of HG⁃induced injury of H9c2 cells was established by CCK8 method.The cells were divided into the control(CON)group and the HG group,which were treated with 5.5 mmol/L and 33 mmol/L glucose respectively.Then the H9c2 cells treated with 33 mmol/L glucose were transfected with si⁃NC,si⁃EZH2,NLRP3⁃NC and NLRP3⁃OE respectively according to the liposome transfection method,and were recorded as group A,group B,group C,group D,group E and group F.Cell apoptosis,EZH2,NLRP3 inflammasome,Cleaved caspase⁃3,Bax protein expression,IL⁃1b and IL⁃18 mRNA expression were detected.Results EZH2,NLRP3,ASC,caspase⁃1 and IL⁃1b,and IL⁃18 mRNA expression in the CON group was lower than those in the HG group(t=6.359,2.941,3.795,4.539,17.874,22.507,P<0.05).Group B was lower than Group HG(t=3.168,6.829,3.639,4.737,12.348,13.915,11.378,P<0.05),Group B was lower than Group A(t=3.727,5.336,8.485,4.859,11.300,9.831,P<0.05),Group D was higher than Group C(t=19.163,10.257,5.743,9.304,16.382,21.493,P<0.05).Compared with the expression of cleared caspase⁃3,Bax protein and apoptosis rate:HG group>A group>B group>CON group,HG group and B group had no statistical significance(P>0.05),and the rest had statistical significance(P<0.05).Compared with group E,apoptosis rate,EZH2,NLRP3,ASC,caspase⁃1,cleared caspase⁃3,Bax protein expression and IL⁃1b and IL⁃18 mRNA in group F were increased(t=11.658,9.212,6.333,10.592,11.457,17.422,13.163,22.763,9.163,P<0.05).Conclusion In the high glucose environment,the expression of EZH2 protein is increased,and silencing EZH2 can target and inhibit NLRP3 inflammasome⁃mediated H9c2 cardiomyocyte apoptosis and inflammatory response injury by regulating epigenetic modification.
作者
代路
高世龙
周彦文
杨青霞
孟馥芬
DAI Lu;GAO Shilong;ZHOU Yanwen;YANG Qingxia;MENG Fufen(Center of Anesthesia and Perioperative Medicine,the Third Clinical Medical College(Affiliated Cancer Hospital),Xinjiang Medical University,Urumqi,Xinjiang,China,830011)
出处
《分子诊断与治疗杂志》
2023年第5期784-788,共5页
Journal of Molecular Diagnostics and Therapy
基金
新疆维吾尔自治区自然科学基金面上项目(2021D01C388)。
