| ObjectivesThe foundation of compound Phyllanthus UrinariaⅡ(CPUⅡ)was based on the theory of TCM and the experience of treating chronic hepatitis and liver fibrosis by screening and optimizing its components for many years.CUPⅡwas composed of ye xia zhu(Phyllanthus Urinaria),ku shen(Sophora flavescens Ait.L),hu zhang(Polygonum cuspidatum Sieb.etZucc),dan shen(radix salviae miltiorrhizae),tian san qi(radix notoginseng),tao ren(semen persecae),tai zi shen(Pseudostellaria heterophyla Pax ex Paxet Hoffm),cai hu(Bupleurum chinense DC)and so on.This compound recipe have been demonstrated that have the better effectiveness of anti-experimental liver injury and liver fibr -osis.This experimental research is to study the influence of CUPⅡon the TGF-β1mRNA Expression in the liver of rats with liver fibrosis,the serum levels of LN,PCⅢand the liver content of oxyproline hydr-oxyproline(Hyp), and study the influence of CUPⅡon the TIMP-1mRNA,BaxmRNA expression in the HSC-T6 and to explore the mechanism of protecting against liver injury at the molecular level by observing the effects of CUPⅡon rats with liver fibrosis induced by DMN and HSC-T6.It aimed to look for the target on inhibiting liver fibrosis of CUPⅡand lay a foundation of treating liver fibrosis with CUPⅡin clinic treatment.Experimental contents1.To study the effect and mechanism of CUPⅡon rats with experiment al liver fibrosis induced by DMN64 SD rats were randomly divided into five groups:normal group,model group,high dosage of CUPⅡ(37.5g/kg),low dosage of CUPⅡ(9.375g/kg),Gan Lixin group(correspond to 12 times of the clinical dose).Each group contained 14 rats except for the control group have 10 rats.On the first day,rats all in groups,except for in normal group,were given a peritoneal injection of 0.05%DMNwith 2ml/kg,consecutive 3 times per week,with 2/3 dose on the first week,thereafter,with whole dose for 3 weeks.All the groups were given the drugs daily for 4 weeks from the onset of the experiment.After that,Excise the eyes to get blood sample,and cut partial right liver lobe which would be fixed with formaldehyde or quick-frozen with liquid nitrogen.The liver function was detected by auto biochemist analytic machine;the serum levels of LN,PCⅢby immunity,and the liver content of oxyproline hydroxyproline(Hyp) were detected,the expression of TGF-β1mRNA in liver tissue were detected with RT-PCR method.We removed the left liver lobe to observe pathological structure with HE and V.G coloring method.The experimental results showed that serum levels of ALT,AST,GGT,TB in high and low dosage groups of CPUⅡwere all obviously lower to that in the model group(P＜0.01),contents of Hyp in liver tissue and LN,PCⅢin serum were remarkable lower than those of the model group(P＜0.01).Hepatic fibrosis histopathological changes were rema rkable alleviated in two dosage groups of CPUⅡcomparing with those of the modal group(P＜0.01)./he rats' liver TGFβ1mRNA expression of high and low dosage groups were all significantly lower to that of model group(P＜0.01). the experiment results indicated that CPUⅡhas a remarkable prophylactic effect aganist liver fibrosis in rats induced by DMN,probably by protecting hepatocyte,inhibiting hyperplasia of collagen and encouraging ECM degrade -ation.2.To study of the effect of Compound Phyllanthus Urinaria(CPUⅡ)on Inhibi -ting proliferation of HSC-T6 and the expression of TIMPmRNA and BaxmRNAHSC-T6 were cultured and divided into 4 groups randomly:The control group,high,middle and low dosage group(Smg/ml,2.5mg/ml,1.25 mg/ml resp -ectively),MTT colormetric assay was used to evaluate the inhibitive effect on HSC-T6,and the expression of TIMP-1 mRNA were detected by using semi-quaniti tative RT-PCR method.The results showed that high,middle and the low dosage groups of CPUⅡall have remarkable inhibitive effect on the proliferation of HSC-T6,the CPUⅡcan decreases the expression level of TIMP-1mRNA and Baxm RNA,comparing with the control group.The experiment results indicated that the proliferation of HSC-T6 can be inhibited by CUPⅡ.CUPⅡpossess the anti -liver fibrosis by inhibiting the proliferation of HSC-T6,accelerating the apoptosis of HSC-T6 and decreasing the expression level of TIMP1mRNA.Conclusion1.CUPⅡpossess the effect of the prophylactic effects of anti-liver fibrosis in rats induced by DMN,the mechanical is related to anti-hepatic cell necrosis inhibit the expression of TGF-β1mRNA.2.CUPⅡhas the strong inhibitive effect against the proliferation of HSC-T6. CUPⅡcan decrease the expression of TIMP-1mRNA to enhance the atabolism of ECM by MMP,and promote the expression of BaxmRNA to accelerate the apoptosis of HSC.
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