Effect of asymmetric dimethylarginine on the activation of hepatic stellate cells and its mechanism.
- Author:
Jin-cheng LI
1
;
Lan CHANG
;
Dong LU
;
De-jian JIANG
;
De-ming TAN
Author Information
1. Department of Infectious Diseases, Xiangya Hospital, Central South University, Changsha 410008, China. ljc.csuxy@yahoo.com.cn
- Publication Type:Journal Article
- MeSH:
Actins;
biosynthesis;
Animals;
Arginine;
analogs & derivatives;
pharmacology;
Cells, Cultured;
Collagen Type I;
metabolism;
Dose-Response Relationship, Drug;
Gene Expression;
drug effects;
Hepatocytes;
cytology;
drug effects;
metabolism;
Male;
NF-kappa B;
metabolism;
Nitric Oxide;
metabolism;
Nitric Oxide Synthase;
metabolism;
RNA, Messenger;
genetics;
metabolism;
Rats;
Rats, Sprague-Dawley;
Reactive Oxygen Species;
metabolism;
Reverse Transcriptase Polymerase Chain Reaction;
Transforming Growth Factor beta;
genetics
- From:
Journal of Central South University(Medical Sciences)
2007;32(3):427-432
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the effect of asymmetric dimethylarginine (ADMA), an endogenous inhibitor of nitric oxide synthase, on the activation of hepatic stellate cells (HSCs) and its mechanism.
METHODS:Primary HSCs isolated from SD rats were cultured and treated with different concentrations (1, 3 or 10micromol/L) of ADMA for various periods (12 approximately 48h). Expression of alpha-smooth muscle actin (alpha-SMA) and synthesis of type-I collagens in HSC were determined. Messenger RNA levels of the transforming growth factor-beta1 (TGF-beta(1)) in the HSCs were determined using RT-PCR. Intracellular reactive oxidant species (ROS) production was measured using oxidant-sensitive fluorescent indicator. Activation of nuclear factor-kappaB (NF-kappaB) was detected by electrophoretic mobility shift assay (EMSA).
RESULTS:ADMA could increase alpha-SMA-positive cells ratio and Type I collagens production of HSCs in a concentration- and time-dependent manner, concomitant with the increase of the TGF-beta(1) mRNA level. Treatment with ADMA (10micromol/L) significantly increased the intracellular ROS production and activated NF-kappaB. Such effects of ADMA on the level of TGF-beta(1) mRNA could be markedly attenuated by pretreatment with antioxidant pyrrolidine dithiocarbamate (25micromol/L).
CONCLUSION:ADMA can induce the HSC activation by increasing TGF-beta(1) expression through ROS-NF-kappaB-dependent pathway. Therefore, ADMA should be a novel and endogenous activator of HSC, which may be involved in the development of liver fibrosis.
