Therapeutic Effects Of AKF-PD On Hepatic Fibrosis Induced By Carbon Tetrachloride In Rats.

Background liver fibrosis is the common approach in the development of a variety of chronic liver disease to cirrhosis of serious consequences. So the anti-liver fibrosis treatment play a key role in prevention chronic liver disease progress to cirrhosis. At present, there is no ideal medicines to treat the liver fibrosis, developed new anti-liver fibrosis drags have more and more been becoming a hot topic. Recently, AKF-PD, a novel pyridine compound, have been proved that it has anti - renal fibrosis, and in vitro experiments have been proved inhibition proliferation of hepatic stellate cell. this animal experiment, aim to observe the therapeutic effect of AKF-PD on progression of hepatic fibrosis in rats and its possible mechanism.Materials and Methods In this study, liver fibrosis was induced by carbon terrachloride（CCL₄）. 100 Sprague-Dawley rats, weighing 180-240 grams, were divided into two groups, that are normal subject group （n=10） and CCL4 treatment group （n=90）. 10 normal rats were given subcutaneous injections of peanut oil 3.0ml kg^-1, 90 CCL4 treatment group received subcutaneous injections of CCl₄ twice weekly in peanut oil (2: 3, 0.3 ml kg^-1) for 6 weeks, 6 weeks later, CCL4 treatment group were randomly subdivided into 4 groups: CCL4 treatment control group（n=15）, AKF-PD treatment group （n=23）、interferonα-2b （IFNα-2b） treatment group （n=14）, AKF-PD+ IFNα-2b group （n=16）. AKF-PD treatment group were given 400mg.kg^-1.d^-1 AKF-PD sodium carboxymethycellulose suspension intragastric administration; interferonα2b group were given 100, 000 units. rat^-1.d^-1 subcutaneous injections; AKF-PD+ IFNα-2b group were given received300 mg.kg^-1.d^-1 AKF-PD sodium carboxymethycellulose suspension intragastric administration in combination with 100, 000 units. rat^-1.d^-1 interferonα2b subcutaneous injections; CCL4 treatment control group、normal group were given 8 ml/kg.d^-1 0.5% sodium carboxymethycellulose suspension intragastric administration, after treatment for 4 weeks, and all rats were put to death, the HE, Masson staining of liver tissue were performed to observe pathological change of liver fibrosis and semi-quantitative analysis was done, Immunohistochemical staining were performed tor detect the expressions of typeⅠcollagen,α-smooth muscle actin （a-SMA）, Transforming Growth Factor betal （TGF-β1） in liver tissue.Results （1） CCL4 treatment control group, when compared to normal group, were seen more fibrous tissue hyperplasia, higher pathology stage of hepatic fibrosis according to HE staining （s3-s4 is 80% in CCL4 treatment control group） （p＜0.01）, higher semi-quantitative score according to Masson staining （p＜0.01）, and more typeⅠcollagen expression according to immunohistochemical staining semi-quantitative analysis （p＜0.01）.（2） AKF-PD group, when compared to CCL4 treatment control group, were seen less fibrous tissue hyperplasia, lower pathology stage of hepatic fibrosis （s3-s4 is 41% in AKF-PD group） （p＜0.05）, less typeⅠcollagen expression （p＜0.05）. But there is no significant difference in the semi-quantitative score of hepatic fibrosis according to masson straining（p＞0.05）.there is no significant difference between AKF-PD treatment group and IFN-α2b-treatment group（p＞0.05）, there is no significant difference between AKF-PD+ IFNα-2b group and CCL4 treatment control group （p＞0.05）, There are no more effective in AKF-PD in combination with IFN-α2b treatment than in AKF-PD only group and IFN-α2b only group （p＞0.05）.（3） CCL4 treatment control group were detected more a-SMA, TGF-β1 expression than normal group （p＜0.01） and AKF-PD treatment group （p＜0.05） according to immunohistochemical staining semi-quantitative score. But a-SMA, TGF-β1 expression is no significant difference between AKF-PD group and IFN-α2b group（p＞0.05）, and between the AKF-PD + IFN-α2b group and CCL4 treatment control group（p＞0.05）. TGF-β1 expression is higher in AKF-PD + IFN-α2b group than in AKF-PD group and IFN-α2b group. But a-SMA expression is no difference between AKF-PD + IFN-α2b group and AKF-PD group and IFN-α2b group.Conclusions （1） AKF-PD has antifibrotic effect on liver in CCL4 model rats, AKF-PD and IFN-α2b treatment may have similar effect on anti-fibrosis of liver in CCL4 model rats. （2） The mechanism of AKF-PD anti-fibrosis effect may be mainly associated with the inhibition of HSC activity, down-regulation TGF-β1 expression, and reduction ECM synthesis. （3） The AKF-PD in combination with IFN-α2b treatment has no more effect than AKF-PD only and IFN-α2b only treatment.