The Expression Of Glucocoticoid Receptor In Liver Failure And The Preliminary Discussion Of Its Mechanism

Objective:To study the expression of glucocorticoid receptor(GR) subtype in acute-on-chronic liver failure and the relationship with the prognosis of patients with ACLF;Through establishment of animal model,to study the molecular mechanism of GR in response to liver failure,and provide ideas for clinical treatment.Methods:1.Collected EDTA anticoagulants from the 26 patients with ACLF who were patients from the first hospital affiliated to soochow university infection department during June 2015 to March 2016,and 10 cases of healthy physical.Using RT-PCR detection the expression of GRa and GRβ in PBMCs,comparing the differences of GRa and GRβin ACLF and HCs,the death and survival groups.Analysis the relationship of GR and prognosis.2.To establish the animal model,35 Balb/c mice were randomly divided into normal control group,chronic liver damage group,hepatic failure group,1mg/Kg methylprednisolone therapy group,2mg/Kg methylprednisolone therapy group.In addition to the normal group,the rest four sets were injected with 20% CCl4 olive dilution intraperitoneally of 5ml/Kg. 2 times per week for 8 weeks.Then the group of 3、4、5 were injected with D-GalN 500mg/Kg and LPS 100 ug/Kg intraperitoneally after 8 weeks.Then the model of ACLF is established.At the same time,the group of 4、5 were treated with 1mg/Kg methylprednisolone and 2mg/Kg methylprednisolone respectively for 5 days.3.Identifiy whether the model is success or not through observeing the liver pathological after HE staining.Using Immunohistochemical and Western blot to detect the expression of GRa and GRβ.At the same time,using Western blot to detect the activation of p38 MAPK signaling pathway,to analysis the relationship of p38 MAPK signaling pathway with the change of GR subtype,and the signaling pathway’s regulation of GR.Result:1.Compared with healthy people,the expression of GRa mRNA is lower than that of the patients of ACLF(P<0.05),while the expression of GRa mRNA in patients of ACLF is higher than that of healthy people(P<0.05).2.The expression of GRa mRNA in the survives of ACLF is higher than that non-survives(P<0.05),while the expression of GRβ mRNA has no obvious difference(P>0.05).3.HE staining:after 8 weeks,there is no death in the 7 rats of group 1,the surface of liver is smooth,the liver cells is complete.There is 2 deaths in group 2,the surface of the liver is rough,the liver cells changed to fatty and fibrosis under the microscopic.There is 3 deaths in group 3,liver tissue is hyperemia swelling,the liver cells changed to large necrosis on the basis of fibrosis,and there is inflammatory cell infiltration around the central vein and bile duct.The liver injury alleviate in group 4 and 5 after hormone therapy.4.Immunohistochemical:GRa of mice liver cells were expressed in 5 groups,and the positive cells of GRa in group of liver failure is the least,group1>group2、4、5>group3,and the positive cells of GRβ in group of liver failure is the maximum.5.Western blot:The trend of GRa and GRβexpression is consistent with immunohistochemical detection in the 5 groups.The expression of phosphorylated p38 MAPK increased in liver failure group,then after hormone treatment,the expression of phosphorylated p38 MAPK decreased,and in paraller with the changes in the GRa.Conclusion:1.The decrease of GRa and increase of GRβ in liver failure group,may be related to inflammatory cytokines in liver failure.The inflammatory cytokines increase the expression of GRβ higer than GRa.The GRβ can combine with GRa into dimers to interfere GRa’s biological activity.2.The expression of GRa mRNA in the survives of ACLF is higher than that non-survives,while the expression of GRβ mRNA has no obvious difference.Then GRa can be used to predict the prognosis of ACLF.3.From the result of HE staining,CCl4 can induce chronic liver damage in mice,combine with D-GalN and LPS can induce acute liver failure,and hormone therapy can improve the degree of liver damage.4.The p38 MAPK signaling pathway activated in liver failure,and participate in the regulation of GR change.The hormone treatment can inhibit the pathway activation to play the role of anti-inflammatory.