Anti-hepatic Fibrosis Effects Of Bifendate In Rat And Its Relationship With CBR1 And FABP1

Objective: Cannabinoid receptors(CBR1) and liver fatty acid-binding protein(FABP1) are considered to be potential therapy targets for hepatic fibrosis. However, the distribution and function of the two receptors in liver tissue are still unknown. In this study, the distributions of CBRs and FABP1 in liver were observed. In the basis, the protective effects of Bifendate(BDD) on hepatic fibrosis induced by CCL4 in rats were analysed. New tagets may be found in the diagnosis and treatment of hepatic fibrosis in this experiment.Methods: 1 Hepatic fibrosis models in rats induced by CCl4 combined with alcohol. 30 Wistar rats were divided randomly into 3 groups of 10 each: normal control group(Control group), CCl4 combined with alcohol model group(CCl4 group) and CCl4 combined with alcohol treatment with Bifendate group(Bifendate group). Bifendate group was administrated with Bifendate by gastric lavage, once a day for 8 weeks. Control group and CCl4 group were given the same volume of 0.5% sodium carboxymethyl cellulose respectively. Meanwhile, Both CCl4 group and Bifendate group were treated with subcutaneous injection of 40% CCl4 solution in peanut oil(0.5ml/100 g for the first time, and then 0.3ml/100 g twice a week for 6 weeks), Control group was given equivalent peanut oil solution. Except the control group, 10% ethanol solution was given ig for 6 weeks.2 Determination of biochemical parameters in serum After 8 week, rats were anesthesiaed, the blood samples was obtained for the assay of AST, ALT, TP and ALB in serum.3 Observation of liver morphologyThe liver were observed grossly and then weighed for calculating the organ index. Some liver tissues were embedded in paraffin and seriallysectioned for HE staining and Masson staining.4 Determination of hepatic hydroxyproline contentsSome hepatic tissues were homogenized for the determination of hepatic hydroxyproline contents using commercially available kits according to the manufacture’s instruments.5 Determination the expression of FABP1 and CBR1The expressions of FABP1 and CBR1 in liver tissues were detected with immunohistochemistry method and western blot method.Results:1 Effects of Bifendate on the index of liver and spleenLiver index of CCl4 group was higher than that of Control group. Liver index of Bifendate group decreased significantly compared with CCl4 group. However, it is still higher than that of Control group.2 Effects of Bifendate on liver functionthe activities of serum AST and ALT were increased in CCl4 group compared with those in control group, while the serum ALB and TP levels were decreased. The elevated levels of AST and ALT in serum in CCl4 group were decreased significantly in Bifendate group. The decreased TP and ALB levels were obviously attenuated in Bifendate group.3 Gross anatomy observation of the liverThe surface of the liver in control group was smooth. The surface of the liver in CCl4 group was rough and granular manifesting as liver cirrhosis. The liver in Bifendate group was better than CCl4 group, no obvious granular substances were found on the surface.4 Histopathological examination of the liveHE staining: The structure of liver lobule was destroyed obviously in the CCl4 group, accompanied with hepatocellular hydropic degeneration and steatosis. Spotty necrosis and infiltration of inflammatory cells were also observed in CCl4 group. The proliferation of fibrous tissue in portal area was observed. Star-like fibrous tissue hyperplasia surrounded the hepatic lobule. In Bifendate group, the structure of liver lobule was almostlycomplete and the structure was clear. Degeneration, necrosis and inflammatory cell infiltration in the hepato-cell are not as serious as thosed in CCl4 group. Some fibrous hypertrophy was observed in portal area.Masson staining: Uneven liver structure was seen in the CCl4 group, different degree of collagen fiber hyperplasia and pseudolobule were formed. In the Bifendate group, the extension of collagen fibers between liver tissues became smaller. No pseudolobule in the livers was seen.5 Determination the contents of hydroxyproline in liverThe hydroxyproline levels in CCl4 group were elevated significantly compare with those in the control group, the hydroxyproline levels after bifendate treatment was decreased compared with CCl4 group.6 Expression of FABP1 in liver tissueFABP1 was presented in Liver cell cytoplasm in Control group, the positive cells were decreased in CCl4 group compared with that of control group, and the level of positive cells after bifendate treatment was increased compared with CCl4 group.7 Expression of CBR1 in liver tissueLittle expression of CBR1 in the liver tissue of Control group was found, while the expression of CBR1 in the liver in CCl4 group was higher than that in control group. The expression of CBR1 in the liver tissue was dercreased after the animal was treated with Bifendate.Conclusions:1 The expression of FABP1 was decreased while the expression of CBR1 were increased in CCl4 combined alcohol induced hepatic fibrosis in rats.2 Bifendate attenuates CCl4 combined alcohol induced epatic fibrosis in rats, which may be related to its effect of enhancement the expression of FABP1 and depressant of the expression of CBR1.