The Influences Of Ganfukang On The Extracellular Matrix And Related Factors Of Experimental Hepatic Fibrosis

Objective To observe the changes of serum HA,LN,CIV,PCⅢand MMP-2,TIMP-2 which are correlated with the extracellular matrix (ECM) degradation after the effect of GFK on experimental rats with hepatic fibrosis. To elucidate the expression patterns of MMP-2, TIMP-2. To determin the potential genetic mechanism of hepatic fibrosis's happen, development and reverse. To provide an adjuvant therapeutic clue for better treatment of human hepatic fibrosis.Methods Healthy SD rats were divided into seven groups at random including normal control group(n=10),Ganfukang high dose group(n=12),Ganfukang medium dose group (n=12),Ganfukang low dose group(n=10),model group(n=11), model control group(n=11), and preventive group (n=11). All groups except normal control group were subcutaneous injected 10% CCl4(5ml/kg) for 12 weeks and two times each week to obtain hepatic fibrosis and were treated with sodium chloride,GFK3125mg/kg ,GFK312.5mg/kg, GFK31.25mg/kg respectively after nine weeks of CCl4 injection. Model control group were treated with sodium chloride. Preventive group were treated with GFK 312.5mg/kg.d for two week as soon as the model is established. At the end of the twelfth week, rats in model and preventive groups were put to death, reserve the serum to detect four ECM targets. Also fetch out the liver as quickly as possible and reserve in the -70℃refrigeratory for the RT-PCR. The mRNA expression of MMP-2 and TIMP-2 were measured by semi-quantitation reverse transcriptase polymerase chain reaction (RT-PCR). The RT-PCR results was input into the image analysis software to detect the integrated option density(IOD)to show expression level and so to analysis and compare. At the end of the twenties week, the left rats were treated with the same methods as the above. RIA technique were applied to detect the serum levels of CIV,PCⅢ,HA,LN.Results 1) The serum level of HA,LN,CIV,PCⅢwere higher in the model and model control groups, which has significant difference compare with normal control group (P<0·05). There is no significant difference between preventive and normal control group (P>0·05). The collagen fibrosis decreased significantly in GFK group, this groups has significant difference compare to model group (P<0·05). And the effect of medium dose GFK is the most effective. 2) RT-PCR demonstrated that expression of MMP-2 and TIMP-2 in model group increased significantly compared with normal control group,While the expression of MMP-2 and TIMP-2mRNA in the therapeutic groups reduced significantly compared with model group. The same result we can get is that the effect of medium dose GFK is the most effective.Conclusion 1) GFK has preferable preventive and therapy effects on rats with hepatic fibrosis. And the effect of medium dose GFK is the most effective. It's the reasonable dosage.2) GFK can significantly relieve the degree of hepatic fibrosis. This may be through the way that GFK inhibit synthesis of collagen and reduce generation of ECM. 3) GFK can probably remove the MMP-2 inhabitor so to improve the bioactivity of MMP-2 to promote the ECM degradation. 4) GFK may play its anti-fibrosis function through regulates gene expression, repress the expression of MMP-2 and TIMP-2 mRNA to make MMP-2 and TIMP-2 tend to be balanced.