Proteomic Analysis of the Effect of Fuzheng Huayu Recipe on Fibrotic Liver in Rats

Abstract

Hepatic fibrosis is a common pathological process of chronic liver diseases and would lead to cirrhosis, and Fuzheng Huayu (FZHY) is an effective Chinese herbal product against liver fibrosis. This study observes FZHY influence on proteome of fibrotic liver with differential proteomic approach and aims to understand FZHY multiple action mechanisms on liver fibrosis. The liver fibrosis models were induced with intraperitoneal injection of dimethylnitrosamine for 4 weeks in rats and divided into model control (model) and FZHY-treated (FZHY) groups, while normal rats were used as normal control (normal). After model establishment, rats in FZHY groups were administered 4 g/kg wt of FZHY for 4 weeks, and normal and model groups were given the same volume of saline. The liver proteins in the above 3 groups were separated by two-dimensional gel electrophoresis (2-DE), the differentially expressed spots were analyzed and compared between normal and model or model and FZHY groups, and then the proteins were identified with mass spectrum analysis and validated partially with western blot and real-time PCR. 1000~1200 spots were displayed on each 2D gel, and a total of 61 protein spots were found with significant intensity difference between normal control or FZHY and model control. 23 most obviously differential spots were excised, and in-gel digestion and 21 peptide mass fingerprints (PMF) were obtained with MALDI-TOF MS analysis, and 14 proteins were identified through protein database searching. Among 14 differentially expressed proteins, 8 proteins in normal and FZHY groups had the same tendency of differential expression compared with the ones in model group. And one of them, vimentin, was validated by western blot and real-time PCR analyses. Our study reveals 12 proteins responsible for fibrogenesis induced by DMN in rats, and among them, 8 proteins in fibrotic liver were regulated by FZHY, including aldehyde dehydrogenase, vimentin isoform (CRA_b), gamma-actin, vimentin, fructose-bisphosphate aldolase B, aldo-keto reductase, S-adenosylhomocysteine hydrolase isoform, and HSP90. It indicates that the action mechanism of FZHY antiliver fibrosis may be associated with modulation of proteins associated with metabolism and stress response, as well as myofibroblast activation. The study provides new insights and data for exploring the liver fibrogenesis pathophysiology and FZHY action mechanism against liver fibrosis.

Potential biomarkers associated with diabetic glomerulopathy through proteomics

“…The biological role of AKR1 levels in diabetic glomerular fibrosis is unclear. Previous study clearly demonstrated that the fibrotic liver had the decreased expression levels of AKR1 when compared to the normal liver 32 indicating that AKR1 responsible for fibrogenesis in liver. Interesting, AKR1 was up-regulated in association with serum AFP, and was an independent risk factor for HCC in chronic hepatitis C patients, suggesting its possible involvement at a very early stage of hepatocarcinogenesis.…”

Role of differential proteomics in studying pharmacological mechanisms of traditional Chinese medicines

“…Fu-zheng-hua-yu (FZHY) is an effective Chinese herbal product that can cure liver fibrosis. A study using 2-DE coupled with MALDI-TOF-MS showed that FZHY regulated the expression of aldehyde dehydrogenase, gammaactin, fructose-bisphosphate aldolase B, aldo-keto reductase, S-adenosylhomocysteine hydrolase isoform, and HSP 90 in a dimethyl nitrosamineinduced rat model of liver fibrosis (37). The results indicate that the molecular mechanism by which FZHY affects liver fibrosis may be through the modulation of proteins involved in metabolism and stress responses, as well as myofibroblast activation.…”

A proteomic analysis of mushroom polysaccharide-treated HepG2 cells

“…For protein identification, the dried spots were analyzed using a REFLEX-III (Bruker) MALDI-TOF-MS 50 . The spectrometer was run in the positive ion mode and in the reflector mode with the following settings: pulsed N2 laser 337 nm, ion source 1 = 19.00 kV, ion source 2 = 16.50 kV, reflector voltage = 20.00 kV, lens voltage = 8.80 kV, pulsed ion extraction time = 80 ns, matrix suppression = 400 Da, and positive reflector mode.…”