Levels of histone modifications in activated primarily cultured rat hepatic stellate cells
10.3969/j.issn.1000-4718.2015.05.018
- VernacularTitle:大鼠原代肝星状细胞活化后组蛋白修饰水平的观察
- Author:
Tian TIAN
;
Jinjuan ZHANG
;
Xinhua LUO
;
Rujia XIE
;
Bing HAN
;
Ting YANG
;
Tengxiang CHEN
;
Qin YANG
- Publication Type:Journal Article
- Keywords:
Hepatic stellate cells;
Histone modifications;
Liver fibrosis
- From:
Chinese Journal of Pathophysiology
2015;(5):871-876
- CountryChina
- Language:Chinese
-
Abstract:
AIM:To investigate the changes of histone modifications during the activation of primarily cultured rat hepatic stellate cells ( HSCs) and the relationship between histone modification patterns andα-smooth muscle actin (α-SMA) expression, and to explore the roles of histone modifications in the activation of HSCs.METHODS:The rat HSCs were isolated by in situ perfusion of collagenase combined with density gradient centrifugation, cultured in vitro and identi-fied by immunofluorescence staining.The morphological features of the cells were observed under inverted microscope.The changes of desmin and α-SMA during the activation of HSCs were detected by immunofluorescence staining and Western blotting.The levels of histone 3 lysine 4 dimethylation (H3K4me2), histone 3 lysine 9 dimethylation (H3K9me2), his-tone 3 lysine 9 acetylation (acH3K9) and histone 4 lysine 12 acetylation (acH4K12) in quiescent HSCs and activated HSCs were determined by Western blotting.RESULTS: The morphology of HSCs shifted from a quiescent phenotype to highly activated myofibroblast during the culture.Immunofluorescence staining and Western blotting showed that the expres-sion levels of α-SMA and desmin were increased over time and reached maximum at 15 d.According to the results of cell morphology and immunofluorescence staining, the cells cultured for 24 h and 15 d were quiescent and activated HSCs, re-spectively.Compared with quiescent HSCs, there were higher H3K4me2 and lower H3K9me2, acH3K9 and acH4K12 modification levels in activated HSCs ( P<0.01 ) .CONCLUSION: Histone modifications show anomalous expression during the activation of primarily cultured rat HSCs.Histone modifications may contribute to the transdifferentiation of HSCs and the development of hepatic fibrosis.
