Effect of salvianolic acid B on apoptosis of liver fibrotic cells based on cleaved caspase-9 protein
10.3969/j.issn.1001-1978.2019.06.018
- Author:
Yue KAN
1
Author Information
1. Dept of Pharmacology, College of Basic Medicine, Anhui Medical University
- Publication Type:Journal Article
- Keywords:
Apotosis;
Cleaved caspase-9;
HSC-T6;
Liver fibrosis;
Salvianolic acid B;
TGF-β1
- From:
Chinese Pharmacological Bulletin
2019;35(6):827-832
- CountryChina
- Language:Chinese
-
Abstract:
Aim: To investigate the effect of salvianolic acid B (Sal B) on liver fibrotic cells in vivo and in vitro from die perspective of apoptosis, as well as the effect on cleaved caspase-9. Methods: Diethylnitrosamine (DEN) was used to induce liver fibrosis in mice for 12 weeks. The padiological changes were detected by HE staining, and the fibrotic lesion area was determined. The cell apoptosis in the fibrotic area was observed by Hoechst 33258 fluorescence staining. The expression of cleaved caspase-9 in fibrotic tissues was detected by Western blot. The apoptotic rate of each group was detected by double standard method AnnexinV-FITC/PI, and the expression of apoptotic protein cleaved caspase-9 in HSC-T6 was detected by Western blot. Results: HE staining suggested that 12 weeks were the period of liver fibrosis in mice. No pseudoplobular structure was formed in group with low and high dose of Sal B, and the degree of fibrosis was lower than that in model group. In the fibrotic lesion area, the fluorescence staining of Hoechst 33258 showed that apoptotic cells significantly increased in group with low Sal B and high dose compared with model group. The results of the AnnexinV-FITC/PI method showed that TGF-β1 inhibited the apoptosis of HSC-T6, and Sal B promoted the apoptosis of HSC-T6 after TGF-β1 intervention and showed a concentration dependence (P <0. 01). Western blot results showed that in fibrotic liver tissues, Sal B increased the expression of cleaved caspase-9 in HSC-T6 cells compared with model group. Compared with TGF-β1 group, Sal B increased cleaved caspase-9 protein expression (P < 0. 01). Conclusions: Sal B can significantly promote apoptosis of liver fibrotic cells in vitro and in vivo, and its pro-apoptosis mechanism may be related to the up-regulation of cleaved caspase-9.
