Study On The Expression Level Of NF-κB Signaling Pathway In Acute-on-chronic Liver Failure

Objective: To investigate the expression of NF-кB in patients with acute-on-chronic liver failure(ACLF)and its relationship with the prognosis of patients.The animal model was used to study the changes of NF-кB p65 in the development and treatment of acute-on-chronic liver failure Its molecular mechanism,for the clinical immune regulation and treatment to provide more scientific basis.Objective:1.A total of 32 patients with chronic hepatitis B(ACLF)who were hospitalized in the Department of Infectious Diseases,First Affiliated Hospital of Soochow University from August 20 to January 2017 were enrolled in this study.Ten patients with healthy volunteers(HD)were treated with RT The expression of NF-кB in mononuclear cells(PBMCs)was detected by PCR,and the differences of NF-кB expression between the two groups were compared.The differences of ACLF expression between NF-кB and the survival group were analyzed.Its relationship with prognosis.2.Forty Balb / c mice were randomly divided into normal control group(group 1),chronic liver injury group(group 2),acute-on-chronic liver failure group(group 3),1 mg / kg methylprednisolone therapy group(group 4).Except that the normal control group,the other three groups were diluted with olive oil into 20% carbon tetrachloride(CCl4)5ml / Kg intraperitoneal injection,2 times / week for 8 weeks,the first 8 weeks later,group 3 and 4 to D-galactosamine(D-GalN)500mg / Kg and endotoxin(LPS)100ug / Kg intraperitoneal injection,the establishment of slow plus acute liver failure model,while the first 4 groups to 1mg / Kg methylprednisolone intraperitoneal injection once a day for 5 days.3.The expression of NF – кB p65 and Cleaved Caspase-3 were detected by immunohistochemistry and Western blot.The results were analyzed by statistical method.The results were analyzed by HE staining.Result:1.Results of clinical trials and prognosis: The expression level of NF-кBm RNA in ACLF patients was significantly higher than that in HD group(p<0.01).The expression level of NF-кBmRNA in ACLF survival group was higher than that in death group,but the difference has no significant statistical significance(p> 0.05).2.Physiological condition and HE staining results of mice: After 8 weeks,the first group of 10 mice without death,liver surface gloss,no swelling,liver cells arranged in the rules,the normal shape;group 2 no death,liver volume increases,rough surface,sand grain,mirror under the occurrence of steatosis and collagen deposition and even fibrosis;group 3 died 2,liver congestion and swelling,redness,a large number of liver cells in the fibrosis based on the emergence of sub-large and large necrosis,portal area visible inflammatory cells Infiltration;group 4 after death by hormone therapy 1,liver tissue congestion redness,microscopic see liver cell arrangement disorder,hepatocyte sub-large or large necrosis,collagen deposition and fibrosis,liver damage than the third Group slightly reduced.3.Immunohistochemical staining results: NF-κB p65 was expressed in all groups,and the positive staining was less common in the normal control group,and most of them were found in the cytoplasm.The positive staining of chronic liver injury group,liver failure group and methylprednisolone group was significant(p <0.01).The expression of NF-κBp65 in methylprednisolone group was lower than that in liver failure groups(p=0.02<0.05).Cleaved Caspase-3 was expressed in each group.The positive staining was found in the normal control group,and most of them were found in the cytoplasm.The positive staining of chronic liver injury group,liver failure group and methylprednisolone group was obvious(p<0.01).The expression of Cleaved Caspase-3 in methylprednisolone group was less than that in the liver failure group and the chronic liver injury group(P >0.05),which was light yellow or brownish or even tan,mostly located in the nucleus.4.Western blot results: NF-κB p65 was expressed in each group,and the expression level of NF-κB p65 was higher in the normal control group than that in the control group(P<0.01).And the expression of NF-κB p65 in methylprednisolone group was significantly lower than that in the group of liver failure(p=0.02<0.05).The expression of Cleaved Caspase-3 was significantly lower in the normal control group than in the control group(P <0.01).The expression level of Cleaved Caspase-3 mRNA in inmethylprednisolone group was no significantly lower than that in chronic group(P>0.05).However,the expression of Cleaved Caspase-3 in methylprednisolone group was significantly lower than that in the group of liver failure(p <0.01).Conclusion:1.Results of clinical trials and prognosis:NF-KB signaling pathway can be activated and take part in the progress of acute-on-chronic liver failure.2.HE pathology staining: CCl4 can successfully induce secondary liver injury in mice.Late combined with D-GalN and LPS can induce acute liver failure on the basis of chronic liver failure,and the use of hormone intervention can improve the degree of liver cell damage and affirm the effect of immunoregulatory therapy.2.Immunohistochemical staining and Western blotting: In the acute and chronic liver damage,the NF-кB signal transduction pathway is activated,and the steroid methylprednisolone can inhibit the conduction of NF-кB signal transduction pathway to a certain extent.In acute and chronic liver damage,exacerbated Caspase-3 to perform hepatocyte apoptosis,the hormone can effect on the apoptosis of hepatocytes to a certain extent.