Molecular Mechanism Of Association Between SNRK And SHBG,An Independent Predictor Of Non-Alchoholic Fatty Liver Disease

Background:It is critical to explore noval serological markers and screening tools for Nonalcoholic fatty liver disease(NAFLD).Bioinformatics techniques have deepened our understanding of the molecular mechanisms and pathophysiological changes of human liver disease.NAFLD is now being defined using a new set of diagnostic criteria and has been designated the novel nomenclature metabolic-dysfunction associated fatty liver disease(MAFLD)in 2020.Sucrose non-fermentative 1-related kinase(SNRK)is an emerging field of human energy metabolism and inflammation.At present,there are no reports on the molecular mechanism of the association between SNRK and sex hormone binding globulin(SHBG),an independent predictor of NAFLD,and the role of SNRK in the regulation of liver energy metabolism and inflammation.This topic is divided into three parts to discuss the molecular mechanism of association between NAFLD/MAFLD and NAFLD independent predictor SHBG and SNRK.1.Correlation between MAFLD and SHBG and the optimal anthropometric indicator for predicting MAFLDObjective:This study intended to validate and compare the MAFLD predictive and diagnostic capability of 8 anthropometric indicators and SHBG.Methods:The study involved a population-based retrospective cross-sectional design.The Fangchenggang area male health and examination survey(FAMHES)was used to collect data of SHBG and eight anthropometric indicators,involving body mass index(BMI),waist-to-height ratio(WHtR),waist-hip ratio(WHR),body adiposity index(BAI),cardiometabolic index(CMI),lipid accumulation product(LAP),visceral adiposity index(VAI),and abdominal volume index(AVI).Receiver operating characteristics(ROC)curves and the respective areas under the curves(AUCs)were utilized to compare the diagnostic capacity of each indicator for MAFLD and to determine the optimal cutoff points.Binary logistic regression analysis was applied to identify the odds ratios(OR)with 95%confidence intervals(95%CI)for all anthropometric indicators and MAFLD.The Spearman rank correlation coefficients of anthropometric indicators,sex hormones,and MAFLD were also calculated.Results:All selected anthropometric indicators and SHBG were significantly associated with MAFLD(P<0.001),with an AUC above 0.785.WHtR(90.68%)and SHBG(77.28%)have the highest diagnostic sensitivity and specificity,respectively.Conclusion:Obesity and SHBG levels were associated with the occurrence and development of MAFLD in males.WHtR was identified as the strongest predictor of MAFLD in young Chinese males.2.Bioinformatics analysis of the relevancy between SNRK and NAFLDObjective:This study aimed to explore the mechanism of SNRK in NAFLD by bioinformatics technology.Methods:R Studio software and R packages were applied to analyze Gene Expression Omnibus database(Data sets including GSE61260,GSE37031 and E-MEXP-3291)by microarray sequencing and single cell RNA sequencing(scRNAseq)transcriptome analysis.Results:SNRK has the highest expression in liver endothelial cells and increased in NAFLD and NASH pathological conditions(Kruskal-Wallis test,P<0.01);SNRK was mainly related to lipid metabolism,extracellular matrix,receptor activity of immune and virus,antigen presentation,chemokine release,threonine phosphorylation,and the migration of leukocyte and monocyte.Conclusion:SNRK is an up-regulated gene in NAFLD and NASH,which related to disease progression.3.The molecular mechanism of association between SNRK and SHBG,an independent predictor of NAFLDObjective:To explore the molecular mechanism of association between SNRK and SHBG,an independent predictor of NAFLD,and the functional role of SNRK in the occurrence and development of NAFLD.Methods:HepG2 cells and paraffin sections were used to determine the expression of SNRK in hepatocytes by immunofluorescence and immunohistochemistry techniques.The NAFLD cell model was successfully established by the intervention of oleic acid in HepG2 cells.SNRK overexpression vector and SNRK shRNA interference vector were used to transfect HepG2 NAFLD model.Western blot and PCR techniques were used to explore the molecular mechanism of association between SNRK and SHBG,and the function and possible mechanism of SNRK in NAFLD.Results:The expression of SNRK was positively correlated with SHBG and PI3K,while negatively correlated with NF-κB,p-mTOR and mTOR.Meanwhile,SNRK was negatively correlated with TNF-α,IL-6,FASN,SCD1,CD36 and PPARγ.Conclusion:SNRK may response to the upregulation of HNF-4α and SHBG by downregulating NF-κB(SNRK/mTOR/NF-κB pathway),inhibit hepatocyte inflammation and de novo hepatic lipogenesis...