| Part ?.S3I-201 exerts antifibrotic effect in CCl4-induced hepatic fibrosis modelObjective:To establish the CCl4-induced hepatic fibrosis model and to investigate the anti-fibrotic effects of S3I-201 in liver.Method:Female C57BL/6J mice with 6-7 weeks were randomly divided into normal control group?CCl4 group?CCl4+2mg/kgS3I-201 group?CCl4+10mg/kgS3I-201 group.From the 5th week of the modeling,CCl4+2mg/kgS3I-201 group ?CCl4+10mg/kgS3I-201 group were injected intraperitoneally of 2,10 mg S3I-201/kg body weight Daily,2 weeks totally.normal control group and CCl4 group were injected intraperitoneally of an equal volume of DMSO.Six weeks later,the mice were killed,and the liver index was calculated.The histological examination(HE staining,Masson staining,and immunohistochemical staining)was used to determine the liver injury and extracellular matrix deposition in the liver.The serum inflammatory cytokines(IFN-?,IL-1?,IL-6,and TNF-?)in each group were detected by ELISA to reflect intrahepatic inflammation.Detection of liver biochemical markers(ALT,AST,ALP)was to define the degree of liver damage in mice.The expression of extracellular matrix(?-SMA,fibronectin)and signal pathway molecules(STAT3,p-STAT3,p65,p-p65)expression in mouse liver were detected by Western blot.To investigate the effects of S3I-201 on STAT3 and NF?B signaling pathways.Result:In the CCl4-induced liver fibrosis model,HE staining showed that the steatosis of the Cl4+10 mg/kg S3I-201 group was significantly improved compared with the CCl4 group.Masson staining showed that collagen fibers in liver tissue of CCl4+2mg/kgS3I-201 group and CCl4+10mg/kgS3I-201 group were significantly reduced compared with CCl4 group.Compared with CCL4 group,the liver body index of 10 mg/kg S3I-201 group was significantly lower.Compared with CCL4 group,the serum transaminase in 10 mg/kg S3I-201 group was significantly decreased.ELISA showed that the four inflammatory factors in the 10mg/kg S3I-201 group were significantly lower than those in the CCL4 group.In addition,the levels of IFN-? and IL-6 in the 2mg/kg S3I-201 group were significantly lower than those in the CCL4 group.Immunohistochemistry showed that the deposition of ?-SMA in the liver of CCl4 group was significantly more than that in the normal control group.Western blot also showed that the content of ?-SMA and fibronectin in CCl4 liver of CCl4 group was significantly higher than that of normal control group.It was also found that the intrahepatic fibronectin levels in the CCl4+2mg/kgS3I-201 group and CCl4+10mg/kgS3I-201 group were significantly lower than those in the CCl4 group.Western blot analysis of hepatic STAT-3 and p65 signaling pathways revealed that both CCl4+2mg/kgS3I-201 and CCl4+10mg/kgS3I-201 groups attenuated p-STAT3 and p-p65 signaling.Conclusions:As a STAT3 inhibitor,S3I-201 can effectively slow down the progression of hepatic fibrosis and at the same time inhibit NF?B to exert anti-inflammatory effects.Part ?.S3I-201 exerts anti-tumor effect on models of cell-line-derived xenografts of Hep G2Objective:To establish models of cell-line-derived xenografts to study the effect of S3I-201 in inhibiting hepatocellular carcinomaMethod:Female BALB/C-nu/nu nude mice with 4-5 weeks were randomly divided into CDX + equal PBS group,CDX + 2 mg/kg S3I-201 group,and CDX + 10 mg/kg S3I-201 group.CDX+2mg/kg S3I-201 group and CDX +10mg/kg S3I-201 group were intraperitoneally injected with 2,10 mg S3I-201/kg body weight once a day from the 6th day after cell transplantation.2 weeks totally.The control group was intraperitoneally injected with PBS and DMSO.The volume of transplanted tumors in each group was observed.All mice were anesthetized on the 24 th day after Hep G2 inoculation.The expression of STAT3,p-STAT3,p65,p-p65 was detected by Western blot,and the effects of S3I-201 on STAT3 and NF?B signaling pathways were investigated.Result:On day 18 of inoculation,10 mg/kg S3I-201 treatment significantly inhibited tumor growth relative to the CDX+equal PBS group.On the 24 th day of inoculation,2mg/kg S3I-201 and 10mg/kg S3I-201 could significantly inhibit tumor growth.Western blot analysis showed that the expression of p-STAT-3 was significantly down-regulated in CDX+2mg/kg S3I-201 group and CDX+10mg/kg S3I-201 group,while the down-regulation of STAT3 was not obvious.In CDX+2mg/kg S3I-201 group and CDX +10 mg/kg S3I-201 group,the expression of p-p65 was significantly up-regulated.Conclusions:As a STAT3 inhibitor,S3I-201 can effectively inhibit the increase of tumor volume and exert an anti-tumor effect by inhibiting the STAT3 signaling pathway. |
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