Effect Of Jiajiansanjiasan On The WNT/?-catenin Pathway And LncRNA And MRNA In Rats With Hepatic Fibrosis Based On The Theory Of "Zhukejiao"

Objective: Observe the effect of Jiajiansanjiasan on hepatic fibrosis rats and its effect on WNT / ?-catenin pathway.The mechanism of its lnc RNA and m RNA is revealed.The modern biological connotation of the theory of "Zhukejiao" is enriched.Methods:1.Experiment one.There ara 40 SPF-level SD male rats,which were randomly divided into blank control group,model group,No.1 group(Jiajiansanjiasan No.1 group),No.2 group(Jiajiansanjiasan No.2 group),and positive control group(colchicine).Routine feeding 7D.Model group,No.1 group,No.2 group and positive control group used subcutaneous injection of carbon tetrachloride oil to manufacture liver fibrosis model.At the same time,each group of molds was given intragastric administration at a dose of 1ml/100 g.The blank control group and the model group was given ultra-pure water,No.1 group was given JJSJS prescription,No.2 group was given JJSJS prescription No.2 and the positive control group was given colchicine.After 56 days,take blood and liver tissue.The pathological morphology was observed by HE.ALT and AST were detected.Detected the expression of WNT2,?-catenin,and GSK-3? in blank control group,model group,No.1 group and positive control group by Western blot method.2.Experiment two.There ara 40 SPF-level SD male rats,which were randomly divided into blank control group,model group,Jiajiansanjiasan(JJSJS)group and positive control group(colchicine).Routine feeding 7D.Model group,JJSJS group,positive control group used subcutaneous injection of carbon tetrachloride oil to manufacture liver fibrosis model.At the same time,each group of molds was given intragastric administration at a dose of 1ml/100 g.The blank control group and the model group was given ultra-pure water,the JJSJS group was given JJSJS and the positive control group was given colchicine.After 56 days,take blood and liver tissue.The pathological morphology was observed by HE.and MASSON.Serum levels of HA,LN,PCIII and CIV were detected by ELISA.High-throughput second-generation sequencing was used to detect the expression profiles of lnc RNA and m RNA in liver tissue of rats in blank control group,model group and JJSJS group.Result: 1.Pathological section: HE staining light microscopy observation:Experiment one:compared with the blank control group,the degree of liver fibrosis in the model group,No.1 group,No.2 group and the positive control group increased significantly(P?0.01);compared with the model group,the degree of liver fibrosis in No.1 group and the positive control group reduced significantly(P ? 0.01)and there was no significant difference in No.2 group(P?0.05).Compared with No.1 group,the degree of liver fibrosis in No.2 group increased significantly(P?0.01).Experiment 2:compared with the blank control group,the degree of liver fibrosis in the model group,the JJSJS group and the positive control group increased significantly(P ? 0.01);compared with the model group,the degree of liver fibrosis in the JJSJS group and the positive control group was reduced significantly(P ? 0.01).Compared with the JJSJS group,the degree of liver fibrosis in the positive control group was no significant difference(P?0.05).MASSON staining light microscopy observation.Experiment 2:compared with the blank control group,the degree of liver fibrosis in the model group increased significantly(P?0.01),the JJSJS group and the positive control group increased(P?0.05);compared with the model group,the degree of liver fibrosis in the JJSJS group and the positive control group reduced significantly(P?0.01).Compared with the JJSJS group,the degree of liver fibrosis in the positive control group was no significant difference(P?0.05).2.Levels of liver function.Experiment 1:compared with the blank control group,the serum ALT and AST levels in the model group,the No.1 group and the No.2 group were significantly increased(P?0.01).The serum ALT and AST levels in positive control group were similar to those in the blank control group and there was no statistical significance(P?0.05).Compared with the model group,the ALT levels in No.1 group,No.2 group and the positive control group were significantly lower(P?0.01).Compared with No.1 and No.2 group,the AST level in No.1 group and No.2 group was reduced(P?0.05),and the AST level in the positive control group was significantly lower(P<0.01).Compared with No.1 group,the level of AST in the positive control group was significantly reduced(P?0.01).3.Pathway.Experiment 1:compared with the blank control group,the expression of WNT2,?-catenin and GSK-3? in the liver tissue of model group decreased significantly(P?0.05),while the expression of GSK-3? decreased significantly(P?0.01);compared with model group,the content of WNT2,?-catenin and GSK-3? in positive control group and No.1 group had no significant change(P?0.05).4.Indicators of hepatic fibrosis.Experiment 2:compared with the blank control group,the serum concentrations of HA,LN,PC III and CIV in the model group were significantly increased(P?0.01).Compared with the model group,the concentration of HA in the JJSJS group increased significantly(P ? 0.05).Compared with the model group,the concentration of LN,PCIII and CIV in the JJSJS group decreased(P ? 0.05).No significant difference in serum HA,LN,PCIII and CIV levels between the JJSJS group and the positive control group(P?0.05).5.Sequencing results.Experiment 2:1976 new lnc RNAs were predicted.There were 340 Inc RNA genes(187 up-regulated and 153 down-regulated)in blank control group and model group.There were 251 Inc RNA genes(139 up-regulated and 112 down-regulated)between the blank control group and the JJSJS group.There were 93 Inc RNA genes(48 up-regulated and 45 down-regulated)in model group and JJSJS group.The main functions of different target genes of lnc RNA are: natural killer cell mediated immunity,natural killer cell mediated cytotoxicity regulation and so on.The main pathways of differential lnc RNA target genes are pentose phosphate pathway and so on.There were 1635 differentially expressed genes(up 1451 and down 184)between the blank control group and the model group,80 differentially expressed genes(up 39 and down 41)between the model group and the JJSJS group,and 80 differentially expressed genes(up 39 and down 41)between the blank control group and JJSJS group.The common pathway of m RNA differential genes in the blank control group,JJSJS group and model group is PPAR signaling pathway.Conclusion:1.The rat model of hepatic fibrosis was successfully reproduced by CCl4 induction.2.The effect of JJSJS No.1 on reducing the degree of liver fibrosis in rats is better than that of in JJSJS No.2 which removing chuanshanjia.3.Jiajiansanjiasan(JJSJS No.1)does not act through WNT/?-catenin pathway in treating liver fibrosis in rats.4.Jiajiansanjiasan changed the expression profile of lnc RNA and m RNA in liver fibrosis rats,which may be part of the mechanism of JJSJS in treating liver fibrosis rats.