| Objectives:To investigate the effects and mechanisms of Fufang Bie Jia Ruan Gan(FBJRG) tables and combination of FBJRG tables and Bifidobaeterium infantis on experimental liver fibrosis.Methods:The effects of FBJRG tables and combination of FBJRG tables and Bifidobaeterium infantis were investigate via three different animal models of liver fibrosis, induced by chemical damage, alcohol and concanavalin, respectively. The research consisted by four parts:(1) Prevention and treatment of FBJRG tables on liver fibrosis induced by carbon tetrachloride(CCl4). In this section, 60 male SD rats were randomly divided into six groups: normal, model, three doses of FBJRG tables-treated groups(including 0.6g/kg, 1.2g/kg, and 2.4g/kg), and colchicine-treated group(0.5mg/kg). The model of liver fibrosis damaged by chemistry was established by subcutaneous injection of CCI4, which procedures were as follows: the pure CCl4 solution was injected at the first time, and then the mixture of 40% CCl4 and olive oil was injected every four days. All rats were administered with corresponding drugs or distilled water by garage. After 8 weeks, the liver weight(LW), liver weight index(LWI), liver function, the concentrations of hyaluronic acid(HA) and precollagen III(PC III) in serum, the concentrations of hydroxyproline(Hyp), matrix metalloproteinase(MMP), tissue inhibitors of metalloproteinases(TIMP), angiotensin II(Ang II), superoxide dismutase(SOD), malondialdehyde(MDA), glutathione peroxidase(GSH-Px) in liver tissues were determined. The hepatic pathological changes and expressions of collagen I/III were evaluated by HE staining and picric-sirius red polatized light method respectively. The Western blot method was used to detect the protein expressions of transforming growth factor-β1(TGF-β1), phosphorylated-smas2/3, smad7 in liver.(2) Prevention and treatment of FBJRG tables on liver fibrosis induced by alcohol. The groups were divided as the same way with part1. The model was established by administration of white spirit once a day in the morning, and the corresponding drugs in the afternoon by gavage. The normal group was applied by distilled water. After 10 weeks, the LW, LWI, liver function, the concentrations of HA and PC III in serum, the concentrations of Hyp, SOD, MDA, GSH-Px, collagen I and collagen III in liver tissues were determined. The hepatic pathological changes were evaluated by HE staining. The relative expressions of CYP2E1 m RNA were observed by real time- RT PCR. The Western blot method was used to detect the protein expressions of transforming growth factor-β1(TGF-β1) and α-smooth muscle actin(α-SMA) in liver.(3) Prevention and treatment of FBJRG tables and combination of FBJRG tables and Bifidobaeterium infantis on immune liver fibrosis. In this section, 80 mice were divided into 8 groups: involving normal, model, three doses groups of FBJRG tables-treated(including 0.9g/kg, 1.8g/kg and 3.6g/kg), Bifidobaeterium infantis-treated group(0.5g/kg), combination of FBJRG tables and Bifidobaeterium infantis-treated group(3.6g/kg BJRG tablets, 0.5g/kg Bifidobaeterium infantis) and colchicine-treated group(0.75 mg/kg). The concanavalin solution(12.5 mg/kg) was given by injection once a week, and the mice were administered with corresponding drugs or distilled water(normal group) by gavage once a day. After 7 weeks, the LW, LWI, liver function, the concentrations of HA and PC III in serum, the concentrations of Hyp, Tumor necrosis factor-α(TNF-α), interleukin-6(IL-6), IL-10, MMP-1, MMP-2, MMR-9, Collagen I/III in mice liver tissue were detected. The m RNA expressions and protein expressions of TGF-β1 were measured as well.(4)The liver physiologic structures of rats were determined by doppler imaging system and ultrasonic real-time tissue elastography.Results:(1) In CCl4 induced rat liver fibrosis model, FBJRG tablets could improve liver function, attenuate liver pathological changes, reduce LW, LWI, the serum concentrations of HA and PC III, the expressions of Hyp, MMP-2, MMP-9, TIMP-1, TGF-β1, p-samd2/3, MDA, collagen I and collagen III in liver tissue enhance the activities of SOD and GSH-Px, and increase the expressions of MMP-1 and smad7.(2) In ethanol induced rat liver fibrosis model, FBJRG tablets could improve liver function, attenuate liver pathological changes, ameliorated oxidative stress, and reduce LW, LWI, the serum concentrations of HA and PC III, the expressions of Hyp, TGF-β1, α-SMA, collagen I and collagen III in liver tissue.(3) In concanavalin induced mouse immune hepatic fibrosis model, FBJRG tablets could improve liver function, reduce LW, LWI, the serum concentrations of HA and PC III, the concentrations of Hyp, collagen I and collagen III in liver tissue. BJRG tablets could decrease the level of MMP-2, MMP-9, TIMP-1 and increase concentrations of MMP-1 in liver tissues. FBJRG tablets could down-regulated the expressions TGF-β1m RNA both in m RNA and protein level. However, FBJRG tablets had no effects on inflammatory factor. Bifidobaeterium infantis could decrease the protein concentrations of IL-6 and TNF-α, meanwhile increase the protein concentrations of IL-10 in liver tissues. The results showed that FBJRG tables may have different anti-liver fibrosis mechanism compared with Bifidobaeterium infantis, and drug combination may be a good choice in the treatment of liver fibrosis.(4)The area% of rats liver in treated groups was much less than that in model groups.Conclusions:(1) Fufang Bie Jia Ruan Gan tables have preventive effects on liver fibrosis, which improve liver function and decrease the overproduction of extracellular matrix. The mechanisms may be related to attenuating the oxidative stress, regulating the expressions of MMPs and TIMPs, and intervening the activation of TGFβ1/smad signaling pathway.(2) Bifidobaeterium infantis has anti-liver fibrosis by immunomodulatory effects, and the drug combination has good synergy on the treatment of liver fibrosis.(3)Ultrasonic real-time tissue elastography is helpful in monitoring of liver fibrosis. |
PDF Full Text Request