首页|BMAL1减轻H2O2诱导的心肌细胞损伤机制研究

BMAL1减轻H2O2诱导的心肌细胞损伤机制研究

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目的 探讨脑和肌肉组织芳香烃受体核转运蛋白的类似蛋白1(BMAL1)通过核因子E2相关因子2(NRF2)调节活性氧(ROS)/NOD样受体热蛋白结构域相关蛋白3(NLRP3)炎症小体通路对过氧化氢(H2O2)诱导的心肌细胞损伤的影响.方法 体外培养H9c2细胞和BMAL1稳定过表达的H9c2细胞,建立H2O2诱导的H9c2细胞损伤模型,并将细胞分为对照(Control)组、H2O2组、BMAL1过表达(BMAL1-OE)组、BMAL1过表达+H2O2(BMAL1-OE+ H2O2)组、BMAL1过表达+NRF2抑制剂(BMAL1-OE+ML385)组、BMAL1过表达+NRF2抑制剂+H2O2(BMAL1-OE+ ML385+H2O2)组.采用CCK-8法检测细胞活力,荧光探针2′,7′-二氯荧光素二乙酸酯检测ROS生成,Western blot检测BMAL1、NRF2和NLRP3蛋白表达,酶联免疫吸附试验法检测白细胞介素(IL)-1β释放.结果 与Control组相比,H2O2组H9c2心肌细胞活力减弱,ROS生成增多,BMAL1和NRF2蛋白表达水平降低,NLRP3蛋白表达水平升高,IL-1β释放增多(P＜0.05);与H2O2组相比,BMAL1-OE+H2O2组H9c2心肌细胞活力升高,ROS生成减少,BMAL1和NRF2蛋白表达水平升高,NLRP3蛋白表达水平降低,IL-1β释放减少(P＜0.05).与BMAL1-OE+H2O2组相比,BMAL1-OE+ML385+H2O2组H9c2心肌细胞活力减弱,ROS生成增多,NLRP3蛋白表达水平升高,IL-1β释放增多(P＜0.05).结论 BMAL1可减轻H2O2诱导的H9c2心肌细胞损伤,其机制可能与NRF2调节ROS/NLRP3炎症小体通路有关.
Mechanism of BMAL1 attenuating H2O2-induced cardiomyocyte injury
Objective To investigate the effect of BMAL1 on H2O2-induced cardiomyocyte injury through NRF2-regulated ROS/NLRP3 inflammasome pathway.Methods H9c2 cells and H9c2 cells with stable over-expressed BMAL1 were cultured and divided into the control group,the H2O2 group,the BMAL1-OE group,the BMAL1-OE+H2O2 group,the BMAL1-OE+ML385 group and the BMAL1-OE+ML385+H2O2 group.All groups were pre-intervened with corresponding inhibitors,and then treated with 0.2 mmol/L H2O2,except for the control group and the BMAL1-OE group.After the intervention,CCK-8 assay was used to measure cell viability,fluorescent probe DCFH-DA was used to measure ROS generation and Western blot assay was used to detect BMAL1,NRF2 and NLRP3 protein expressions.ELISA was used to determine IL-1β release.Results Compared with the control group,the cell viability was decreased,ROS generation was increased,BMAL1 and NRF2 protein expressions were decreased,NLRP3 expression and IL-1β release were increased in the H2O2 group(P＜0.05).Compared with the H2O2 group,the cell viability was increased,ROS generation was decreased,BMAL1-OE and NRF2 protein expressions were increased,NLRP3 expression and IL-1β release were decreased in the BMAL1-OE+H2O2 group(P＜0.05).Compared with the BMAL1-OE+H2O2 group,the cell viability was decreased,ROS generation was increased,NLRP3 expression and IL-1β release were increased in the BMAL1-OE+ML385+H2O2 group(P＜0.05).Conclusion BMAL1 attenuates H2O2-induced H9c2 cardiomyocyte injury,and its mechanism may be related to the regulation of ROS/NLRP3 inflammasome pathway through NRF2.

ARNTL transcription factorsNF-E2-related factor 2reactive oxygen speciesNLR family,pyrin domain-containing 3 proteinBMAL1inflammasome

易娜、肖雯、田源、袁李礼

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长沙市第四医院心内科(邮编 410007)

湖南师范大学生命科学学院

湖南省脑科医院心内科

ARNTL转录因子类 NF-E2相关因子2 活性氧 NLR家族,热蛋白结构域包含蛋白3 脑和肌肉组织芳香烃受体核转运蛋白的类似蛋白1 炎症小体

湖南省教育厅科研项目湖南省卫生健康委科研项目湖南省卫生健康委科研项目湖南省自然科学基金湖南省自然科学基金湖南省中医药科技项目

21B00882022030129602021030105102022JJ800622022JJ80101D2022024

2024

10.11958/20231027

天津医药
天津市医学科学技术信息研究所

天津医药

CSTPCD
影响因子：1.107
ISSN：0253-9896
年,卷(期)：2024.52(2)
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