C Liu1, X Wang, P Liu. 1. Institute of Liver Disease, Shanghai University of TCM, Shanghai (200032).
Abstract
OBJECTIVE: To investigate the main serapharmacological mechanism of antifibrotic action of Fuzheng Huayu 319 Decoction (FHD) at cellular and molecular level. METHODS: Hepatic stellate cells (HSC) were isolated from the normal rats and subcultured. Normal rats were medicated with FHD and their blood serum collected to prepare the medicated serum, which was incubated with the subcultured HSC. Then, the extra- and intracellular collagen synthetic rates were assayed with collagenase digestion method, type I collagen content in the medium measured with ELISA, transforming growth factor beta 1 (TGF beta 1) protein production semiquantified by immunocytochemistry stain and image analysis system, the gene expressions of type I procollagen and TGF beta 1 analyzed by RT-PCR. RESULTS: FHD medicated serum could obviously inhibit the extra- and intracellular collagen synthetic rates, decreased type I procollagen mRNA expression and its secretion, downregulate TGF beta 1 mRNA and its protein expression. CONCLUSION: FHD could inhibit HSC activation markedly, the main mechanism of its antifibrotic action may be the medicated serum inhibition on type I procollagen and TGF beta 1 gene expression.
OBJECTIVE: To investigate the main serapharmacological mechanism of antifibrotic action of Fuzheng Huayu 319 Decoction (FHD) at cellular and molecular level. METHODS: Hepatic stellate cells (HSC) were isolated from the normal rats and subcultured. Normal rats were medicated with FHD and their blood serum collected to prepare the medicated serum, which was incubated with the subcultured HSC. Then, the extra- and intracellular collagen synthetic rates were assayed with collagenase digestion method, type I collagen content in the medium measured with ELISA, transforming growth factor beta 1 (TGF beta 1) protein production semiquantified by immunocytochemistry stain and image analysis system, the gene expressions of type I procollagen and TGF beta 1 analyzed by RT-PCR. RESULTS:FHD medicated serum could obviously inhibit the extra- and intracellular collagen synthetic rates, decreased type I procollagen mRNA expression and its secretion, downregulate TGF beta 1 mRNA and its protein expression. CONCLUSION:FHD could inhibit HSC activation markedly, the main mechanism of its antifibrotic action may be the medicated serum inhibition on type I procollagen and TGF beta 1 gene expression.