Effect Of Dendnobium Mixture In Order Cycle Method On IKK/NF-κB/IκB Signaling Pathway In Diabetic Rats With Hepatic Fibrosis

ObjectiveTo observe the effect of DMOC on the expression of IKK/NF-κ B/I κ B signaling pathway related proteins in diabetic rats with hepatic fibrosis,and to provide experimental and theoretical basis for the prevention and treatment of diabetes with hepatic fibrosis.MethodsAfter 7 days adaptive feeding,70 SPF Wistar rats(weight 200 ± 10g,male)randomly divided into Normal control group(NC,n=10,fed with normal diet)and model estabalishded group(n=60,according to the random number table.The model estabalishded group was fed with high fat and high sugar diet.After 6 weeks,the model estabalishded group was given intraperitoneal injection of STZ {25 mg/(kg + m2),72 h intervals},twice.And the normal control group Intraperitoneally injected of normal saline water.Selecting 50 T2DM-formulated rats were randomly divided into Model group,DMBG group.DMOC group,DMOC1 group,and DMOC2 group and each group were 10 rats.They were injected subcutaneously with CCl4(2 ml/kg twice a week),when gavage simultaneously,the first dose doubled.On the third day afer the diabetes model success.each group was start intragastrically administered.Each group was weighed once a week,and the injection and gavage doses were adjusted according to body weight every week.At the 8 weeks of CCl4 injection and intragastric administration.blood was drawn from the abdominal aorta.ALT and AST were detected by an automatic biochemical analyzer.HA,LN,PC-Ⅲ,and C-Ⅳ were detected by radioimmunoassay.HE and Masson staining were performed on the liver,and the amount of collagen-Ⅳ in liver tissue was measured by immunohistochemistry.Western blot was used to detect the protein expression of IKK β,NF-κ B P50,NF-κ B P65,I κ B β and p-I κ B β in the liver.Results1 General observation of rats:NC group rats were generally in good condition,Model group was generally in poor condition,and the rest of the treatment groups were significantly improved compared to the Model group.2 Fasting blood glucose changes in rats:Compared with the NC group before the drug intervention,the fasting blood glucose of all groups increased significantly,and and ≥11.1 mmol/L(P<0.01);Model group,DMBG group.DMOC group,and DMOC1 group There was no difference in fasting blood glucose between the two groups(P>0.05).After drug intervention,there was no significant difference between the drug groups(P>0.05).Compared with the Model group,each drug group had fasting blood glucose decreased significantly(P<0.01).3 Liver function changes in rats:After treatment.AST and ALT were higher in each treatment group than in NC group(P<0.01).Compared with Model group,AST and ALT were significantly lower in each treatment group,and there were significant differences(P<0.01),there was no significant difference between the treatment groups(P>0.05);and the ALT decreased more significantly in the DMOC group.4 Four changes of liver fibrosis in rats:After treatment.HA,LN.PC-Ⅲ,and C-Ⅳ in the Model group were significantly higher than those in the NC group(P<0.01).Compared with the Model group,HA,LN.and There ere significant differences between PC-Ⅳ and C-Ⅳ.Four total curative effects on liver fibrosis.5 HE staining of rat liver:After treatment,compared with the NC group,the liver tissue structure of the Model group changed significantly,there was a large number of connective tissue proliferation and collagen deposition,the normal structure was almost completely destroyed,the liver cells were arranged disorderly,and the liver Cellular fatty degeneration is more serious,there are a lot of fat vacuoles,the hepatic cord structure almost disappears,with a large number of inflammatory cell infiltration;compared with the Model group,the hepatic cord structure in the DMBG group has been restored,the hepatocyte structure has been improved,and the Shiqi mixture has been improved.The degree of steatosis,inflammatory changes and structural damage in the group were significantly improved.The connective tissue hyperplasia and collagen deposition in the DMOC group were more improved than those in other groups.The hepatic sinusoids were significantly reduced.6 Masson staining of rat liver tissue:After treatment,compared with the Model group,the collagen fibers in the treatment groups were significantly reduced,and the mediastinum was significantly thinner.And the fibrous tissue deposition was the least,the mediastinum was the smallest,and the fibrosis curative effect was reduced optimal in the DMOC group.7 Collagen-IV immunohistochemistry of rat liver tissue:In the Model group,a large number of Collagen-IV(light yellow)deposits were found in extracellular hepatocytes,which were reduced in all treatment groups,and the DMOC group had the least yellowish area,in the DMOC pair decreased.Collagen-IV deposition has the best therapeutic effect.8 Expression of IKK β,I κ B β,P-I κ B β,NF-κ B P65,and NF-κ B P50 in liver tissue of rats:Compared with the NC group,expression levels of NF-κ B P65,NF-κB P50,and IKKβ in the Model group increased,and I κ B β,P-The expression of I κ B β was decreased,and there was a significant difference(P<0.01).Compared with the Model group,the expression of NF-κ B P65,NF-κ B P50 and IKK β was decreased in all treatment groups,and the expression of I κ B and P-I κ B β was increased.Difference(P<0.01).Conclusion1 High fat and high glucose + STZ combined with CCl4 can successfully establish an animal model of diabetes with liver fibrosis.2 DMOC can improve blood glucose,liver function index(ALT.AST),liver fibrosis index(HA.LN.PC-III,C-IV),and can reduce collagen and collagen fiber deposition in rat liver of diabetic liver fibrosis rats induced by high fat and high glucose + STZ combined with CCl43 DMOC may inhibit the hepatic fibrosis in diabetic rats by down-regulating the expression of NF-κ B P65,NF-κ B P50 and IKK β and up-regulating the expression of Iκ Bβ and p-I κ Bβ by acting on IKK/NF-κ B/I κ B signaling pathway.