Study on the Mechanism of Tianxiangdan Capsules Alleviating Hypoxia-Reoxygenation Injury in H9C2 Cells by Regulating Pyroptosis Through ERα
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.
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