Experimental Study On Prophylactic Treatment Effect Of AKF-PD On Hepatic Fibrosis Induced By Carbon Tetrachloride In The Rats

Background and Objective: Hepatic fibrosis can be classified as a wound healing response to a variety of chronic stimuli. Hepatic fibrosis involves a mismatched balance of production and degradation of extracellular matrix, mainly collagens. Hepatic fibrosis, which may ultimately lead to cirrhosis, is the pathological base of all the chronic hepatic diseases. Hepatic fibrosis is a reversible disease, but cirrhosis is irreversible. Therefore, it is of practical significance to seek a medicine that can inhibit or delay the progression of hepatic fibrosis. AKF-PD is a novel pyridine compound which is synthesized by our own study group. In our previous study, it has been proven that it can ameliorate renal fibrosis in vivo and in vitro, and it can inhibit proliferation of HSC in vitro. So we decided to observe the prophylactic treatment effect of AKF-PD on CCL4-induced experimental hepatic fibrosis in rats , and to explore some mechanisms of AKF-PD to reduce the hepatic fibrosis.Materials and Methods: Eighty-four SD rats were divided randomly into five groups: Normal group(n=10)was injected with 3ml/kg peanut oil subcutaneously twice a week and intragastric administration of CMCNa 6ml/kg per day; Model group(n=20)was injected subcutaneously with 400ml/L CCL4 in peanut oil at a dose of 3ml/kg twice a week(the first dosage was 5ml/kg)and intragastric administration of CMCNa 6mL/kg per day; AKF-PD prevent group(n=20) was injected subcutaneously with 400ml/L CCL4 in peanut oil at a dose of 3ml/kg twice a week(the first dosage was 5ml/kg)and intragastric administration of AKF-PD 300mg/kg per day; IFN-αprevent group(n=20) was injected subcutaneously with 400ml/L CCL4 in peanut oil at a dose of 3 ml/kg twice a week(the first dosage was 5ml/kg) and intra-muscular injection of IFN-α2b in saline daily at the dose of 1×10~5U; AKF-PD+IFN-a prevent group(n=14)was injected subcutaneously with 400ml/L CCL4 in peanut oil at a dose of 3ml/kg twice a week(the first dosage was 5 ml/kg), intra-muscular injection of IFN-a 2b in saline daily at the dose of 1×10~5U and intragastric administration of AKF-PD 300mg/kg per day. Six weeks later, all rats of each group were killed. Examinations of serum ALT, AST, Tp and Alb were performed, at the same time, tissue sections either were stained with HE, Masson method or subjected to immunohistostaining by using antibodies against collagen I orα-SMA.Results:(1) The model of rat hepatic fibrosis induced by CCL4 was successfully established. Compared with the normal group: the serum level of ALT and AST rised obviously, and the serum albumin decreased in the model group (P＜0.05) .The model group rats liver stained with HE and Masson showed inflammation and necrosis, and extensive accumulati -on of collagens ,the grade of hepatic fibrosis was S3 or S4 by using Scheuer system. The semi-quantity determination of collagen I was obviously increased in the model group (P<0.05).(2) The rats liver function and the degree of hepatic fibrosis were ameliorated in AKF-PD prevent group (P<0.05). The grade of hepatic fibrosis was S1 or S2 by using Scheuer system and the semi-quantity determination of collagen I was obviously decreased in AKF-PD prevent group (P<0.05). However there was no obvious difference between AKF-PD and IFN-αprevent group. There was no obvious difference between model group and AKF-PD+IFN-αprevent group.(3) Compared with the normal group, the expression of theα-SMA obviously increased in the model group, and obviously decreased in AKF-PD prevent group compared with the model group (P<0.05). However there was no obvious difference between AKF-PD and IFN-αprevent group. There was no obvious difference between model group and AKF-PD+IFN-a prevent group.Conclusions: These results demonstrated that AKF-PD can prevent hepatic fibrogenesis caused by chronic CCL4 treatment, and the effect may be realized through decreasing the expression ofα-SMA in the liver, and inhibiting the activation of HSC. Combination prophylactic treatment with AKF-PD and IFN-αwas not better than prophylactic treatment with AKF-PD or IFN-αalone on the effect of preventing hepatic fibrogenesis in this animal trial.