Effect of Danggui Shaoyao San Drug-containing Serum on Expression of p-MLCⅡ and MLCⅡ Protein in HSC-T6 Cells Induced by ET-1
10.13422/j.cnki.syfjx.20190226
- VernacularTitle: 当归芍药散含药血清对ET-1诱导的HSC-T6细胞内p-MLCⅡ，MLCⅡ蛋白表达的影响
- Author:
Sha-sha JIANG
1
;
Yong-fu PAN
1
;
Mo YANG
1
;
Yun-lai WANG
1
;
Dan-dan YIN
1
;
Fan XU
1
Author Information
1. School of Pharmacy, Anhui University of Chinese Medicine, Key Laboratory of Chinese Medicine Formula of Anhui Province, Hefei 230012, China
- Publication Type:Research Article
- Keywords:
Danggui Shaoyao San;
drug-containing serum;
HSC-T6 cells;
endothelin-1(ET-1);
p-myosin light chain(MLC)Ⅱ
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2019;25(2):14-19
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effect of endothelin-1 (ET-1) on the expression of phosphorylated myosin light chain Ⅱ(p-MLCⅡ)and myosin light chain Ⅱ(MLCⅡ)protein in rat hepatic stellate cells HSC-T6 and explore the intervention effect of Danggui Shaoyao San(DSS)drug-containing serum. Method:After HSC-T6 cells were seeded, DMEM and blank rat serum with final concentrations of 2.5%, 5%, 10%, 15% and 20% were added to each well. The viability of HSC-T6 cells was determined by methyl thiazolyl tetrazolium(MTT) assay to screen the suitable serum concentration range. The cells were divided into blank serum control group (5%, 10%, 15%) and DSS drug-containing serum group (5%, 10%, 15%). ELISA was used to detect the content of ET-1 in cell culture supernatant under basic state. The cells were divided into blank serum control group (10%), DSS drug-containing serum low (5%), medium (10%) and high dose (15%) groups. Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR) was used to detect the level of ET-1 mRNA in cell culture supernatant under basic state. The cells were divided into blank serum control group (10%), model group (10%), DSS drug-containing serum low (5%), medium (10%), high dose (15%) groups and Y-27632 inhibitor group (100 μmol·L-1). Except the blank serum control group, the other groups all received 10 nmol·L-1 ET-1 to induce HSC-T6 cells. Western blot was used to detect the expression of p-MLCⅡ and MLCⅡ in HSC-T6 cells induced by ET-1. Result:Serum concentrations of 5%, 10% and 15% were used as drug-containing serum concentrations. As compared with the blank serum control group, the DSS drug-containing serum group significantly reduced the relative content of ET-1 and ET-1 mRNA in the basic state (P<0.05, P<0.01). As compared with the blank serum control group, the expression of p-MLCⅡ and MLCⅡ protein in the model group was significantly increased (P<0.01); DSS drug-containing serum groups and Y-27632 inhibitor group can significantly down-regulate p-MLCⅡ and MLCⅡ protein expression (P<0.05, P<0.01). Conclusion:DSS drug-containing serum may down-regulate the expression of p-MLCⅡ and MLCⅡ by down-regulating the content of ET-1 and inhibiting the autocrine of ET-1.
