MG132对缺氧/复氧大鼠心肌细胞的保护作用及其与核因子-κB信号通路的相关性  

Roles of NF-κB signaling pathways in MG132 induced reduction of hypoxia/reoxygenation injury to cardiomyocytes

作　　者：李囡[1] 孔祥照[1] 古同男[1] 骆雨辰 鄢雯[1] Li Nan;Kong Xiangzhao;Gu Tongnan;Luo Yuchen;Yan Wen(Clinical Medicine Department,YanJing Medical College,Capital Medical University,Beijing,101300,China)

机构地区：[1]首都医科大学燕京医学院临床医学学系,北京101300

出　　处：《贵州医药》2022年第2期171-173,179,共4页Guizhou Medical Journal

基　　金：首都医科大学自然科学基金(PYZ19143);首都医科大学燕京医学院科研培育基金(2016QD02)。

摘　　要：目的探讨免疫蛋白酶体抑制剂MG132对缺氧/复氧(H/R)条件下心肌细胞的保护作用及其与核因子-κB(nuclear factor kappa-light-chain-enhancer of activated Bcells,NF-κB)信号通路的相关性。方法分离培养新生SD大鼠心肌细胞,建立心肌细胞H/R模型。将心肌细胞随机分为对照组、MG132组、H/R组和H/R+MG132组。应用CCK-8比色法检测各组细胞增殖与活力,用二硝基苯肼法测定培养液上清中乳酸脱氢酶(LDH)活性,用TUNEL法检测细胞凋亡,RT-PCR法测定细胞中肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)和白细胞介素6(IL-6)mRNA水平,Western blot法测定细胞中p-IκB和p-P65的蛋白水平。结果与对照组相比,H/R组细胞活力降低,LDH活性和细胞凋亡增加,IL-1β、IL-6及TNF-αmRNA水平升高,p-IκB和p-P65蛋白表达升高,差异有统计学意义(P<0.05);与H/R组相比,H/R+MG132组细胞活力明显提高,LDH活性和细胞凋亡率降低,IL-1β、IL-6和TNF-αmRNA水平降低,p-IκB和p-P65蛋白表达降低,差异有统计学意义(P<0.05)。结论心肌细胞的H/R过程能激活NF-κB,IL-1β、IL-6及TNF-α的表达也相应增加,导致细胞损伤;而MG132能抑制NF-κB的活化,继而降低NF-κB调控的IL-1β、IL-6及TNF-α的表达,减轻炎症反应,改善H/R损伤。Objective To investigate the protective effect of MG132 on the hypoxia/reoxygenation(H/R)injury to cardiomyocytes as well as its relationship to the nuclear factor kappa-light-chain-enhancer of activated B cell(NF-κB)signaling pathway.Methods The neonatal rat primary cardiomyocytes were cultured in DME/F-12 culture medium supplemented with 10%fetal bovine serum and divided into four groups using a random number table,which were control group,H/R group,H/R+MG132 group and MG132 group.After H/R,The cell viability was measured by CCK-8 assay.The apoptosis was analyzed by TUNEL stain.The lactate dehydrogenase(LDH)activity in the supernatant was evaluated by 4-dinitrophenylhydrazine method.The expression of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and IL-6 at mRNA levels were determined by RT-PCR,respectively.The protein levels of p-IκB and p-P65 in the cells was determined by western blot(WB)assay.Results Compared with control group,cell viability was significantly repressed,and LDH activity and apoptotic rate as well as the expression of IL-1β,IL-6,TNF-αmRNA,p-IκB and p-P65 protein all increased in group H/R(P<0.05).While MG132 alleviated H/R injury when compared with group H/R,as evidenced by the significantly increased cell viability,and further proved by the suppressed LDH activity and apoptotic rate as well as the expression of IL-1β,IL-6 and TNF-αmRNA,p-IκB and p-P65 protein all decreased(P<0.05).Conclusion The H/R process of cardiomyocytes can activate NF-κB,L-1β,IL-6,TNF-α,causing cell damage,while MG132 inhibits activation of NF-κB,and then reduces the expression of L-1β,IL-6,TNF-α,and reduce inflammatory and improve H/R damage.MG132 showed protective effect on the cardiomyocytes with H/R by a possible NF-κB signaling pathway.

关 键 词：心肌细胞 MG132 缺氧复氧 NF-κB 炎症因子 

分 类 号：R34[医药卫生—基础医学]