The effects of curcumin derivatives C66 on proliferation and activation of rat hepatic stellate cells induced by transforming growth factor-β
10.3760/cma.j.issn.1000-6680.2017.08.009
- VernacularTitle:姜黄素衍生物C66对转化生长因子-β刺激的大鼠肝星状细胞增殖和活化的研究
- Author:
Ze CHEN
1
;
Dazhi CHEN
;
Yunlei XIAO
;
Xixi XIAO
;
Yonglin WANG
;
Lanman XU
;
Yongping CHEN
Author Information
1. 325025,温州医科大学附属第一医院感染科温州医科大学肝病研究所温州市肝病重点实验室
- Keywords:
Curcumin derivative;
Hepatic stellate cell;
Liver fibrosis;
c-Jun N-terminal kinase
- From:
Chinese Journal of Infectious Diseases
2017;35(8):492-497
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effects of curcumin derivatives (C66) on proliferation and expressions of α-smooth muscle (α SMA) and Collagen Ⅰ in rat hepatic stellate cells (HSC) induced by transforming growth factor-β (TGF-β) in vitro and the relationship with cannabinoid receptor type 1 (CB1).Methods To determine the optimum time and concentration of C66,HSC-T6 cell line was cultured in vitro and divided into control group and groups with different doses of C66 (1 μmol/L,2 μmol/L,5 μmol/L,10 μmol/L,20 μmol/L).Cell proliferation was detected by Cell Counting Kit-8 assay.Then,according to the time and concentration of C66 above,cells were divided into 5 groups including control group,TGF-β only group,TGF-β combined with CB1 antagonist group,TGF-β combined with C66 group and TGF β combined with CB1 antagonist plus C66 group.Quantitative realtime polymerase chain reaction and western blot were used to assess the expressions of α SMA,Collagen Ⅰ,CB1,JNK and phosphorylation of JNK (p-JNK).The variance homogeneity of multiple samples was compared by LSD method.The variance was compared with Dunnett T3 test.One-way analysis of variance was performed to compare the mean values among the groups.Results The inhibitory effect of C66 on HSC-T6 proliferation was dose and time dependent.The optimum time and concentration were 48h and 10 μmol/L,respectively,with the inhibition rate of 54％.Compared with control group,expressions of α-SMA,collagen Ⅰ and CB1 were significantly elevated in TGF-β group (t=6.188,3.48 and 20.64,respectively,all P＜0.05).TGF-β1 could increase the relative mRNA expressions of CB1,collagen Ⅰ and α-SMA with significant differences (t =4.705,9.492 and 38.27,respectively,all P＜ 0.05).Compared with control group,p-JNK expression was significantly elevated in TGF-β group (t=9.567,P＜0.05).Conclusions C66 could inhibit the proliferation and collagen synthesis in HSC-T6 induced by TGF-β and the effect is strengthened when combined with CB1 antagonist,which may involve JNK phosphorylation.Our study provides a better understanding on the mechanism and a new target for treatment of liver fibrosis.
