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

作　　者：荣幸 姜林 岳远佳 王慧敏 吉钊 陈良 RONG Xing;JIANG Lin;YUE Yuan-jia;WANG Hui-min;JI Zhao;CHEN Liang(The Fourth Clinical Medical College of Xinjiang Medical University,Urumqi 830000,China;State Key Laboratory of the Causes and Prevention of High Morbidity in China,Urumqi 830054,China;Xinjiang Key Laboratory of Traditional Chinese Medicine Processing,Urumqi 830000,China)

机构地区：[1]新疆医科大学第四临床医学院,新疆乌鲁木齐830000 [2]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 [3]新疆中药炮制研究重点实验室,新疆乌鲁木齐830000

出　　处：《中药材》2024年第2期443-447,共5页Journal of Chinese Medicinal Materials

基　　金：国家自然科学基金资助项目(81960699);省部共建中亚高发病成因与防治国家重点实验室开放课题基金资助项目(SKL-HID-CA-2020-8)。

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。Objective:To investigate the effect of Tianxiangdan capsules on hypoxia-reoxygenation(H/R)injury of H9C2 cells through ERαregulating NLRP3.Methods:Cardiomyocytes were cultured in vitro and divided into blank control group,model group,estrogen(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-1βand 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 expression of estrogen receptor ERαand 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,IL-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 increased,the levels of ROS,LDH,IL-1βand IL-18 and the expression of pyroptosis-related proteins were significantly decreased(P<0.05),the red fluorescence of PI decreased significantly,ERαprotein expression of high concentration of Tianxiangdan capsules drugcontaining serum group increased significantly(P<0.05).AZD9496 reversed the effect of Tianxiangdan capsules.Conclusion:Tianxiangdan 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 

分 类 号：R285.5[医药卫生—中药学]