天香丹胶囊通过ERα调节细胞焦亡减轻H9C2细胞缺氧复氧损伤的机制研究
Study on the Mechanism of Tianxiangdan Capsules Alleviating Hypoxia-Reoxygenation Injury in H9C2 Cells by Regulating Pyroptosis Through ERα
荣幸 1姜林 2岳远佳 1王慧敏 1吉钊 1陈良2
作者信息
- 1. 新疆医科大学第四临床医学院,新疆 乌鲁木齐 830000;省部共建中亚高发病成因与防治国家重点实验室,新疆 乌鲁木齐 830054
- 2. 新疆医科大学第四临床医学院,新疆 乌鲁木齐 830000;新疆中药炮制研究重点实验室,新疆 乌鲁木齐 830000
- 折叠
摘要
目的:探讨天香丹胶囊通过ERα调节NLRP3对H9C2细胞缺氧复氧损伤的影响.方法:体外培养心肌细胞,分为空白对照组、模型组、雌激素(E2)阳性对照组及天香丹胶囊含药血清低(5％)、高(15％)浓度组和天香丹胶囊含药血清(15％)+AZD9496(雌激素抑制剂)组、天香丹胶囊含药血清(15％)+MCC950(NLRP3抑制剂)组.CCK-8测定细胞活力;DCFH-DA荧光探针测定心肌细胞活性氧(ROS)水平;ELISA检测细胞上清中LDH、IL-1β、IL-18水平;Hoechst33342/PI双染评价细胞焦亡情况;Western Blot检测雌激素受体ERα和焦亡相关蛋白NLRP3、Caspase-1、GSDMD的表达.结果:与空白对照组比较,模型组细胞活力显著降低,ROS、LDH、IL-1β、IL-18水平及焦亡相关蛋白表达显著升高(P＜0.05),PI红色荧光明显增强.与模型组比较,天香丹胶囊含药血清低、高浓度组细胞活力显著升高,ROS、LDH、IL-1β、IL-18水平及焦亡相关蛋白表达显著降低(P＜0.05),PI红色荧光明显减弱,天香丹胶囊含药血清高浓度组细胞ERα蛋白表达显著升高(P＜0.05).AZD9496可逆转天香丹胶囊的作用.结论:天香丹胶囊可缓解心肌细胞缺氧复氧后损伤,减轻炎症反应,可能与调节ERα和NLRP3焦亡机制有关.
Abstract
Objective:To investigate the effect of Tianxiangdan capsules on hypoxia-reoxygenation(H/R)injury of H9C2 cells through ERα regulating NLRP3.Methods:Cardiomyocytes were cultured in vitro and divided into blank control group,model group,es-trogen(E2)positive control group,low(5％)and high(15％)concentration of Tianxiangdan capsules drug-containing serum groups,Tianxiangdan capsules drug-containing serum(15％)with AZD9496(estrogen inhibitor)group,Tianxiangdan capsules drug-containing serum(15％)with MCC950(NLRP3 inhibitor)group.Cell viability was determined by CCK-8 assay.DCFH-DA fluorescent probe was used to measure the level of reactive oxygen species(ROS)in cardiomyocytes.The levels of LDH,IL-1β and IL-18 in cell supernatant were detected by kit.Hoechst33342/PI double staining was used to evaluate the pyroptosis.Western Blot was used to detect the expres-sion of estrogen receptor ERα and pyroptosis-related proteins NLRP3,Caspase-1,GSDMD.Results:Compared with the blank control group,the cell viability was significantly decreased,and the levels of ROS,LDH,IL-1β,1L-18 and pyroptosis-related protein expression were significantly increased(P＜0.05),PI red fluorescence was obriously enhanced in the model group.Compared with the model group,the cell viability of low and high concentration of Tianxiangdan capsules drug-containing serum groups was significantly in-creased,the levels of ROS,LDH,IL-1β and IL-18 and the expression of pyroptosis-related proteins were significantly decreased(P＜0.05),the red fluorescence of PI decreased significantly,ERα protein expression of high concentration of Tianxiangdan capsules drug-containing serum group increased significantly(P＜0.05).AZD9496 reversed the effect of Tianxiangdan capsules.Conclusion:Tianx-iangdan capsules alleviate hypoxia/reoxygenation injury and reduce inflammatory response in cardiomyocytes possibly by regulating ERαand NLRP3 pyroptosis.
关键词
天香丹胶囊/心肌缺血/心肌焦亡/ERα/NLRP3信号通路Key words
Tianxiangdan capsules/Myocardial ischemia/Myocardial pyrodeath/ERα/NLRP3 Signaling pathway引用本文复制引用
基金项目
国家自然科学基金(81960699)
省部共建中亚高发病成因与防治国家重点实验室开放基金(SKL-HID-CA-2020-8)
出版年
2024
国家科技期刊平台
NSTL
万方数据