June Green Saponins Hepatic Fibrosis And Its Experimental Study Of The Molecular Mechanisms

Objective: To establish the method of determination of saponins in Liuyueqing (LYQS). Methods: The LYQ was extracted with the 60% ethanol, liposoluble impurity was removed by petroleum ether, then extracted by water-saturated butanol. Panaxsaponin Re was used as standard in the preparation of calibration curve. the content of LYQS was detected by Vanillin-CH3COOH-HClO4. Results: The RSD of precision and constancy were 1.35% and 1.25%,the recovery was 100.42%; the yield of LYQS was 4.57%, and the purity was 46.06%.Conclusions: The method is simple, accurate, high precision and constancy. It can be used for the quantitative determination of LYQS. The yield and purity of LYQS are good. Objective: To study the effect of LYQS against CCl4-induced liver fibrosis in rats and its mechanism.Methods: Wistar rats were divided into two groups randomly: hepatic fibrosis model group and normal control group.The rats of Hepatic fibrosis model group were established by intragastric administration (i.g.) of 50%CCl4 for 7 weeks, and the rats in the normal control group, normal saline (NS) were given too. 50 Wistar rats of hepatic fibrosis group that confirmed by the pathological inspection were divided into 5 groups randomly.All rats were treated with drugs or NS for four consecutive weeks by i.g,one time a day.24 hours after the last administration of drugs, all rats were killed.(1)The blood serum and hepatic tissue were taken, and the activities of ALT, AST in serum and the contents of Hyp, SOD, MDA, GSH-PX in hepatic tissue were measured.(2) Pieces of hepatic tissues in the identical spot of liver, size 1×1×l cm3, were taken, were soaked in 10% neutral formalin solution for the light microscope observation.(3) The expressions of Col-I, MMP-2, TIMP-1, TIMP-2, and TGF-β1 mRNA in hepatic tissue were determined by RT-PCR.Results:(1)Compared with the model control group, the activity of ALT in serum was decreased significantly in all LYQS groups and positive control group (P <0.01) ;and the activity of AST was decreased in the positive control group , high dose group and middle dose group (P<0.01); the content of SOD,GSH-PX in hepatic tissue was increased significantly in all LYQS groups and positive control group (P<0.05 or P<0.01) ;and the content of Hyp,MDA was decreased significantly in the meantime (P<0.05 or P<0.01).(2) The pathological results of HE staining showed that, compared with the model control group, the rank of hepatic fibrosis in rats of LYQSH, LYQSM and colchicines was significantly lower(P<0.05 or P<0.01).(3) The expressions of Col-I, TIMP-1 and TIMP-2 mRNA were down-regulated by all dose of LYQS; TGF-β1 mRNA were down-regulated and MMP-2 mRNA was up-regulated by LYQH, LYQM and colchicine (P<0.05 or P<0.01). Conclusions: LYQS might have certain curative effect on CCl4-induced liver fibrosis in rats,its mechanism may be related to attenuating free radical, inhibiting the lipid peroxidation, reducing the collagen synthesis and the deposition, decreasing the sedimentation of ECM and increasing the degradation of ECM. Objective: To investigate the effects and mechanism of LYQS on the proliferation, cytotoxicity and inhibition of fiber synthesis of HSC-T6. Methods: inhibitive effect of LYQS. Three different concentrations of LYQS in which 90 % HSC-T6 cells were survival were prepared. HSC-T6 were treated with LYQS, LDH assay was used to evaluate the cytotoxicity of LYQS, the content of Hyp in the supernatant of cells was detected by Hyp kit, and the expressions of Col-I,TGF-β1,MMP-2,TIMP-1and TIMP-2 mRNA were determined by RT-PCR.Results: LYQS could obviously inhibit the proliferation of HSC-T6 (P<0.05), and the cytotoxicity to the HSC-T6 cell is low; the content of Hyp in the supernatant of cells was reduced by each concentration of LYQS; the expressions of TGF-β1, TIMP-1 and TIMP-2 mRNA were down-regulated by each concentration of LYQS; Col-I mRNA were down-regulated and MMP-2 mRNA was up-regulated by LYQH, LYQM (P<0.05 or P<0.01). Conclusions: LYQS could inhibit the proliferation of HSC-T6, reduce the collagen synthesis, decrease the sedimentation of ECM and increase the degradation of ECM, it might be one of its mechanism of anti-hepatic fibrosis.