| Background and Aims This study was to investigate the role of IL-22 in inhibiting hepatic fibrosis induced by HSC and to explore the role of Wnt/?-catenin pathway in the activation of hepatic stellate cells.It is further clarified whether IL-22 inhibits Wnt/?-catenin Pathways play a role in anti-hepatic fibrosis.Methods Rat HSC-T6 was activated by TGF-?1,The expression of?-catenin and ?-SMA mRNA in HSC was detected by RT-PCR and Western-blot respectively after 48 h of HSC was treated with TGF-?1 at 0,2.5 ng/mL and 5ng/mL protein levels.The proliferation of cells was detected by CCK8 method after intervention with HSC for 48 h at 0,250 pg/mL,500 pg/mL,750 pg/mL and 1000 pg/mL of IL-22,respectively.The proliferation and apoptosis of cells were detected by IL-22 at 1000 pg/mL for 12 h,24 h,36 h,48 h,CCK8 and flow cytometry.The HSC proliferation rate was measured after intervention with IL-22 for 15 h at a dose of 1000 pg/mL for 15 h.Then HSC was pretreated with5 ng/mL TGF-?1 for 24 h and then treated with IL-22 at a dose of 1000 pg/mL for HSC 48 H,the cell proliferation rate was detected again,and the cell proliferation rate was inhibited by IL-22 before and after activation of HSC.Theexpression of ?-catenin,?-SMA mRNA and protein in the cells were detected by IL-22 at a dose of 1000 pg/mL for 48 h.The cells were pretreated with 5 ng/mL TGF-?1 for 24 h and then treated with 1000 The expression of ?-catenin,?-SMA mRNA and protein were detected by IL-22 at a dose of pg/mL for 48 h.The changes of mRNA and protein expression of HSC ?-catenin and ?-SMA were compared before and after intervention,the mRNA and protein levels of?-catenin and ?-SMA were detected by q-PCR and western-blot,respectively.Results The expression of ?-catenin and ?-SMA mRNA and protein increased with the increase of TGF-?1 concentration(P <0.05),suggesting that activation of Wnt/?-catenin pathway and ?-SMA expression changes involved in the process of HSC activation.The proliferation rate of HSC decreased with the increase of IL-22 concentration,and the cell proliferation rate was significantly greater than 0 pg/mL at 750 pg/mL(P <0.05),and the same concentration of IL-22(P <0.05).The apoptosis rate of HSC increased with the increase of IL-22 concentration,but there was no significant difference in apoptosis rate between each group(P > 0.05).Similarly,there was no significant difference in the apoptosis rate between each time group(P > 0.05).Compared with the two groups of IL-22 intervention before and after,the proliferation rate of HSC was significantly decreased after intervention,and the difference was statistically significant(P <0.05).Compared with the two group of IL-22 intervention before and after TGF-?1 activation,the proliferation rate of HSC cells after TGF-?1activation was significantly inhibited by IL-22(P <0.05).The expression of?-catenin and ?-SMA mRNA and protein in HSC cells before and after TGF-?1activation were compared and the results showed that the inhibitory effect in the expression of ?-catenin and ?-SMA mRNA and protein in activated HSC cells was more significant(P <0.05).Conclusion This study indicates that the Wnt/?-catenin pathway may participates in the process of HSC activation and ?-SMA secretion,and IL-22 inhibits biological function of HSC in a dose-and time-dependent manner.This effect probably via inhibited the Wnt/?-catenin signal pathway.The results of this study deepen the understanding of the role of IL-22 against liver fibrosis,for the future development of new anti-liver fibrosis drugs provide experimental basis. |
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