| Objective:This research by studying the expression of LXRs and related factors SREBP-1,Foxp3,RORyt,IL-17,TNF-α in liver tissue of patients with primary biliary cholangitis(PBC)and their relationship with intrahepatic inflammation,to preliminarily explore the role of LXRs in the regulation of PBC inflammation,and by analyzing the levels of Foxp3 and RORyt,the characteristic molecular markers of Treg and Thl7 cells,we explore the correlation between the LXRs and Th17/Treg ratios in liver tissue of PBC patients,to provides new ideas for the study of early cell and molecular events of PBC and the development of innovative treatment strategiesMethods:Collecting liver biopsy specimens from 29 patients who were diagnosed as PBC by clinical,serum biochemical,autoantibody examination and liver puncture pathology examination in the Second Affiliated Hospital of Kunming Medical University from October 2015 to May 2019(PBC group),while collecting 15 liver biopsy specimens from patients who were hospitalized and diagnosed with chronic hepatitis B during the same period(CHB group)and 17 cases of normal liver tissues from the Forensic Medicine School of Kunming Medical University were identified to exclude other medical behaviors and liver diseases(NC group).Immunohistochemical technique was used to detect the expression of LXRs,SREBP-1,RORyt,Foxp3,IL-17,TNF-α,RORyt/Foxp3 in the liver tissues of PBC group,CHB group and NC group;according to the results of HE staining,PBC patients were staged according to different histopathological manifestations,and the levels of the above factors in PBC patients with different pathological stages were compared;the relationship between the expression of LXRs and SREBP-1,RORyt,Foxp3,IL-17,TNF-α,RORyt/Foxp3 was analyzed;the correlation between the expression of LXRs,SREBP-1,RORγt,Foxp3,IL-17,TNF-α,RORyt/Foxp3 and the serum biochemical indexes reflecting the liver function level was analyzed to further explore the relationship between LXRs and intrahepatic inflammation of PBC.Results:1.HE staining resultsAccording to the results of HE staining,the PBC group was staged by histopathology.The results showed that there were 7 cases in phase Ⅰ(cholangitis)in the PBC group,4 cases in phase Ⅱ(inflammation around the junction),and stage Ⅲ(progressive fibrosis)a total of 14 cases,a total of 4 cases of stage Ⅳ(cirrhosis).2.Results of immunohistochemical detection of LXRα,LXRβ,SREBP-1,RORγt,Foxp3,IL-17,TNF-α,RORγt/Foxp3 expression in the liver tissues of PBC group,CHB group and NC group1)The expression of LXRa and LXRβ:LXRa and LXRβ are positive expression in the three groups of subjects,mainly nuclear expression.Compared with NC group and CHB group,the expression levels of LXRa and LXRβ in liver tissues of PBC group are significantly lower(P<0.05),and there is no significant difference between CHB group and NC group(P>0.05);2)The expression of SREBP-1:SERBP-1 is positively expressed in the three groups of subjects,mainly cytoplasmic expression.There was no statistically significant difference in the level of SREBP-1 in liver tissues of PBC group,CHB group and NC group(P>0.05);3)The expression of RORyt:RORyt showed weak positive expression or even no expression in the NC group,and is strongly expressed in the CHB group and PBC group,mainly nuclear expression.Compared with the NC group,the expression of RORyt in the PBC group and the CHB group showed an upward trend.the difference was significant(P<0.01).The difference in the RORyt level between the CHB group and the PBC group was not significant(P>0.05);4)The expression of Foxp3:Foxp3 is expressed in the liver tissues of the three groups of subjects.The cytoplasm or nucleus of cells expressing Foxp3 positive is stained brown or brownish yellow.The expression of Foxp3 in liver tissues of PBC group and CHB group is significantly lower than that of NC group,and the difference is statistically significant(P<0.05),but the difference of Foxp3 level between CHB group and PBC group is not statistically significant(P>0.05);5)The expression of IL-17 and TNF-α:IL-17 and TNF-α are expressed in the three groups of subjects,but weakly or not expressed in the NC group,strongly expressed in the PBC group and CHB group,mainly based on cytoplasmic expression.The expressions of IL-17 and TNF-α in liver tissues of PBC group and CHB group are significantly higher than that of NC group,and the differences are significant(P<0.001).The difference between PBC group and CHB group is not significant(P>0.05);6)The expression of RORyt/Foxp3:The expression of RORyt/Foxp3 in liver tissues of PBC group and CHB group are significantly higher than that of NC group,and the differences are significant(P<0.05).There is no significant difference in the RORγt/Foxp3 level between the PBC group and the CHB group(P>0.05).3.Comparison of levels of LXRα,LXRβ,SREBP-1,RORγt,Foxp3,IL-17,TNF-α,RORyt/Foxp3 in liver tissues of PBC patients with different pathological stages29 patients with PBC were divided into 2 groups according to HE staining results,a total of 11 patients in the early group(including stages Ⅰ and Ⅱ),and a total of 18 patients in the late group(including stages Ⅲ and Ⅳ).Comparing the expression of LXRα,LXRβ,SREBP-1,RORγt,Foxp3,IL-17,TNF-α and RORyt/Foxp3 in different disease stages of PBC,the results showed that compared with the early group,LXRa,LXRβ,Foxp3 expression are significantly reduced,IL-17 and RORyt/Foxp3 expression are significantly increased,the difference is significant(P<0.05).The difference of other factors in the early and late PBC groups are not significant.4.Correlation analysis between LXRs and SREBP-1,RORyt,Foxp3,IL-17,TNF-α,RORyt/Foxp3 in the liver of PBC patientsThe correlation between LXRs and SREBP-1,RORyt,Foxp3,IL-17,TNF-α,RORyt/Foxp3 is analyzed,the results showed that LXRa is related to Foxp3、RORyt/Foxp3(P<0.05),among them,LXRa has a positive correlation with Foxp3,correlation coefficient r=0.460;LXRa has a negative correlation with RORyt/Foxp3,correlation coefficient r=-0.381.There have a correlation between LXRβ and TNF-α(P<0.05),showing a negative correlation,with a correlation coefficient of r=-0.416.6.The correlation between the expression of LXRa,LXRβ,SREBP-1,RORyt,Foxp3,IL-17,TNF-α,RORγt/Foxp3 and the level of liver function in patients with PBCIn order to study the correlation between the expression of LXRa,LXRβ,SREBP-1,RORyt,Foxp3,IL-17,TNF-α,RORγt/Foxp3 and the level of liver function of PBC patients,we also collected clinical biochemical indicators such as ALT,AST,ALP,GGT,TBIL,DBIL,ALB,GLB,A/G,etc.The analysis results showed that only SREBP-1 have a correlation with A/G(P<0.05),showing a positive correlation,the correlation coefficient r=0.380.Conclusions:1.Compared with the control group,the expression of LXRs in liver tissues of PBC patients is low,and the expression in early patients is significantly higher than that in late patients,suggesting that LXRs may be involved in the occurrence and development of PBC.2.In PBC,the expression of Foxp3 in liver tissues is decreased,the expression of RORyt,RORyt/Foxp3 and IL-17 are increased,and the balance of Th17/Treg is disordered.3.The expression of Foxp3 in the liver tissues of PBC patients is reduced,and the expression in advanced patients is significantly lower than that in early patients,suggesting that with the increase of inflammatory activity,Treg cells are suppressed.4.In PBC,the expression of LXRa decreases,the ratio of RORyt/Foxp3 increases,and there have a negative correlation between LXRa and RORyt/Foxp3,suggesting that LXRα may be an important factor affecting Th17/Treg balance in PBC patients,but the specific mechanism needs further study.5.Compared with the control group,the expression of SREBP-1 in the liver tissue of PBC patients did not change significantly. |
