Effect Of Yiqi Huayu Huatan Yangyin Formula On Expression Of PparΓ And DLK1in Rats With Liver Fibrosis

Objective:To evaluate the anti-fibrotic efficacy of Chinese medicine formulawhich consists of four general methods of “benefiting qi, removing bloodstasis, expelling phlegm, nourish yin(Yiqi Huayu Huatan YangyinFormula,YHHY)”, and observe its influence on the expression ofperoxisome proliferator-activated receptor γ(PPARγ) and Delta-like1(DLK1) in rats with liver fibrosis.Methods:Liver fibrosis model was induced by injecting40%(v/v) carbontetrachloride dissolved in olive hypodermically and fed with a kind of“low-protein, high-fat with liquor” diet for the first two weeks.The survived rats after modeling were randomly divided into liver fibrosismodel group(Group M), colchicine group(Group C), low-,moderate-and high-dose of YHHY group(Group LT, Group MT and Group HT). And10rats were recruited as normal control group(Group N). Rats in normalcontrol group and model group were given saline by gavage (3ml/100g), while the other groups were administered orally with colchicinesolution(0.01mg/100g) or different dosage of YHHY herbal decoction(1g/100g,2g/100g,4g/100g), respectively. After the corresponding treatmentfor12weeks, liver inflammation was observed by HE staining, liverfibrosis was graded under Masson staining, and liver ultrastructure wasobserved by transmission electron microscope. The expression of PPARγmRNA was measured by RT-PCR. The protein levels of PPARγ and DLK1were analyzed by Western blot.Results:1. Effect of Yiqi Huayu Huatan Yangyin Formula on liverhistopathology in rats with liver fibrosis.(1) HE staining: Severe steatohepatitis and fibrosis, with theinflammation of interlobular areas was found in the fibrosis modelgroup．Compared with the model group,the extent of steatohepatitis andfibrosis of moderate-, high-dose of YHHY group and colchicine groupwere dramatically decreased,while there was no significant differencebetween the model group and low-dose of YHHY group.(2) Masson staining: Compared with the normal control group, themodel group showed a large amount of collagen deposition with fiberintervals. Liver fibrosis was inhibited in the colchicine group, moderate-and high-dose of YHHY group in deferent degrees. The liverhistopathology of the low-dose of YHHY group was much similar to that of the model group.2. Influence of Yiqi Huayu Huatan Yangyin Formula on ultrastructurein liver of rats with liver fibrosis.The hepatocytes were normal in the normal control group, and therewere abundant mitochondria, endoplasmic reticulum, ribosome andcholangiole microvilli． However, The model group and thelow-dose of YHHY group showed a large number of lipid droplets withoncotic and decreased organelle, and mixed inflammatory infiltration withlymphocytes and fibroblast could be seen in this two groups. The othertreated groups seemed much similar to the normal control group.3. Effect of Yiqi Huayu Huatan Yangyin Formula on expression ofPPARγ and DLK1in rats with liver fibrosis.(1) The expression of PPARγ: Compared with the model group, PPARγmRNA and protein expression was up-regulated in the colchicine group,moderate-and high-dose of YHHY group, while there was no significancedifferences between the model group and the low-dose of YHHY group.(2) The expression of DLK1: Two soluble forms of Dlk1proteins(DLK1-L and DLK1-S) were expressed in injured livers but were nearlyundetectable in healthy rats. DLK1-L was down-regulated in the colchicinegroup, moderate-and high-dose of YHHY group.Conclusion:1. Yiqi Huayu Huatan Yangyin Formula can intervene the expressions of PPARγ and DLK1,which may play an important role in the process ofanti-fibrotic efficacy, promoting liver histological reconstruction andfunctional recovery.2. Yiqi Huayu Huatan Yangyin Formula inhibit liver fibrogenesismaybe in a dose-dependent manner.