Qishen Huoxue granules containing serum can improve the damage of septic cardiomyocytes by inhibiting excessive autophagy / 中国医师杂志

ABSTRACT

Objective:To investigate the inhibitory effect of Qishen Huoxue granule containing serum on excessive autophagy of cardiomyocytes (H9c2) in septic rats and its protective effect on septic cardiomyocytes.Methods:Twelve SPF grade Wistar rats were gavaged with low, medium and high doses of Qishen Huoxue granule [equivalent to crude drugs 12.7, 25.4 and 50.8 g/(kg·d)]. The cultured rat embryonic cardiomyocytes (H9c2) were divided into five groups the control group was cultured with DMEM containing 10% fetal bovine serum (FBS); lipopolysaccharide (LPS) group was treated with DMEM containing 10% FBS+ 1 μg/ml LPS; LPS+ Qishen Huoxue granule low, medium and high dose groups were pre intervened with DMEM containing 10% low, medium and high dose intragastric drug containing serum for 4 h, and then added 1 μg/ml LPS. After co-cultured for 4 h, the mRNA and protein expression of autophagy markers Beclin-1, ATG5 and LC3B were detected by real time fluorescence quantitative polymerase chain reaction (qRT-PCR) and Western blot; After 8, 12, 24 and 48 hours of co culture, the cell activity of cells in each group at different time points was detected by cell counting method (CCK-8) method.Results:The expression of autophagy markers Beclin-1, ATG5 mRNA and Beclin-1, ATG5 and LC3B protein in LPS group increased in the early stage (4 h), which was statistically significant compared with the control group ( P<0.05). However, LPS+ Qishen Huoxue granule groups could reduce the overexpression of Beclin-1, ATG5 and LC3B mRNA and protein caused by LPS ( P<0.05). CCK-8 method showed that the cell activity of LPS group LPS+ Qishen Huoxue granule low, medium and high dose groups decreased significantly at 12 h, 24 h and 48 h, which was significantly different from that of the control group ( P<0.05); The cell activity of LPS+ Qishen Huoxue granule medium dose group at 24 and 48 hours was significantly higher than that of LPS group ( P<0.05). Conclusions:The drug-containing serum of Qishen Huoxue granules at different concentrations had inhibitory effects on LPS-induced autophagy of cardiomyocytes, and the drug-containing serum obtained by intragastric administration of Qishen Huoxue granules in rats can improve the myocardial injury caused by sepsis by inhibiting autophagy.