Qishen Huoxue granules containing serum can improve the damage of septic cardiomyocytes by inhibiting excessive autophagy
10.3760/cma.j.cn431274-20210421-00467
- VernacularTitle:芪参活血颗粒含药血清通过抑制过度自噬改善脓毒症心肌细胞损伤
- Author:
Huan TANG
1
;
Zheng WANG
;
Guoxing WANG
Author Information
1. 首都医科大学附属北京友谊医院急诊科 100050
- Keywords:
Sepsis;
Cardiomyocyte;
Autophagy;
Qishen Huoxue granules;
Containing serum
- From:
Journal of Chinese Physician
2021;23(10):1466-1471
- CountryChina
- Language:Chinese
-
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.
