| Aims 1.To explore the relationship between circulating VEGF-B and NAFLD in the general public.2.To explore the relationship between plasma VEGF-B level and blood pressure,renal function,glucose and lipid metabolism.Methods 1.We recruited NAFLD subjects and non-NAFLD subjects from the Physical Examination Center of Wuhan Union Hospital.The diagnosis of NAFLD is mainly based on liver ultrasound results,supplemented by other examination results and disease history.2.Homeostasis model assessment of insulin resistance(HOMA-IR),body mass index(BMI),HbA1 c,liver function,kidney function and indexes of metabolic syndrome(blood pressure,fasting plasma glucose,fasting lipids)were evaluated.Plasma VEGF-B was detected by enzyme-linked immunosorbent assay(ELISA)assay.3.We use T test and covariance analysis to analyze the differences between the two groups.We use Spearman correlation analysis and multivariate analysis to analyze the correlation between indicators.Results1.Plasma VEGF-B levels were significantly higher(P = 0.022)in subjects with NAFLD(40.11±16.93pg/ml)compared to those without NAFLD(34.73±14.69pg/ml),and analysis of covariance confirmed this result.2.VEGF-B showed a positive correlation with ?-glutamyl transpeptidase(?-GT)and HOMA-IR in univariate analysis(q = 0.242;P = 0.001;q =0.174;P = 0.019,respectively).Multiple linear regression analysis indicated that ALT and ?-GT were independently correlated with VEGF-B even after adjustment for age and gender(q = 0.286;P = 0.01;q =0.237;P = 0.033,respectively).3.Plasma VEGF-B showed a powerful correlation with blood pressure and renal dysfunction.Conclusions 1.Plasma VEGF-B might be a novel clinical parameter associated with NAFLD.2.VEGF-B could be a suitable biomarker for the early detection of hypertension and renal dysfunction.Aims 1.To explore the expression of VEGF-B in human,mouse and cell models of non-alcoholic fatty liver disease(NAFLD).2.To explore the role of VEGF-B in NAFLD.Methods 1.We download the GSE89632 data set from the GEO database,which contains 24 subjects in the control group and 39 subjects in the NAFLD group.Use the Network Analyst online website to analyze the differential genes between the two groups,and analyze the difference of VEGF-B between the two group.2.db/db mice and control mice were fed with normal diet,each group have 5 subjects.We sacrificed the mice at 16-week-old and take their liver tissue.We stained the liver tissue with Oil Red O and H&E staining to detect the lipid deposition and TG content in liver.We explored the expression of VEGF-B in liver tissue and the difference between the two groups by Immunofluorescence staining.qPCR and WB were used to detect the differences of lipid metabolism,endoplasmic reticulum stress,apoptosis and other related indicators,as well as VEGF-B expression between the two groups.3.Human hepatoma cells HepG2 and human monocytic leukemia cells THP-1 were cultured in vitro and treated with palmitic acid(PA)to establish fatty liver model.We detected lipid deposition by Oil red O staining and intracellular TG detection.qPCR and WB were used to detect the differences of lipid metabolism,endoplasmic reticulum stress,apoptosis and other related indicators,as well as VEGF-B gene expression between the two groups.4.We constructed VEGF-B siRNA and interfered HepG2 cells with VEGF-B siRNA for 48 h.qPCR and WB were used to detect whether the expression of VEGF-B is decreased.We detected lipid deposition by Oil red O staining and intracellular TG detection.qPCR was used to detect the differences of lipid metabolism,endoplasmic reticulum stress,apoptosis and other related indicators between the two groups.5.Intervention of HepG2 cells with recombinant human VEGF-B protein(rh-VEGF-B)for 48 h.We detected lipid deposition by Oil red O staining and intracellular TG detection.qPCR was used to detect the differences of lipid metabolism,endoplasmic reticulum stress,apoptosis and other related indicators between the two groups.Results 1.Analyzing the GSE89632 data set,there are a total of 353 different genes between the healthy control population and the NAFLD population.The expression of VEGF-B gene in the NAFLD population was higher than that of the normal control population.2.Liver lipid deposition in db/db mice was more obvious than that in control mice at 16-weeks-old.Immunofluorescence staining results showed that hepatocytes and macrophages in liver can express VEGF-B,and the expression of VEGF-B in hepatocytes and macrophages in db/db mice was higher than that in the control mice.The qPCR and WB results showed that the expression of PPARr?SREBP1?ACO?FASN?CD36?FATP4 and VEGF-B genes in the liver of db/db mice was higher than that of the control mice,while the BCL2 expression was lower than that of the control mice.3.We established fatty liver cells model with PA treatment in vitro,the lipid deposition of HepG2 was significantly increased in PA treatment group.The qPCR and WB results showed that the expressions of PPARr?SREBP1?FASN?ACC?CD36?ATF6?BAX and VEGF-B were higher in the PA treatment group than the control group,and the expression of BCL2 was lower than the control group.In THP-1 cells,the expression of VEGF-B in the PA treatment group was higher than that in the control group.4.The intracellular lipid deposition was reduced in HepG2 which intervened by VEGF-B siRNA.The qPCR and WB results showed that siRNA intervention was success and the expression of VEGF-B was significantly reduced.The expression of PPARr,SREBP1,CD36,FATP1,FATP3 and BAX were decreased in the VEGF-B siRNA intervention group.5.The intracellular lipid deposition was increased in HepG2 which interfered with rh-VEGF-B.The qPCR results showed that the expression of PPARr,FATP1,FABP1 and ATF6 was higher in HepG2 treated with rh-VEGF-B than that of the control group,while the BCL2 expression was lower than that of the control group.Conclusions 1.The expression of VEGF-B is higher in the NAFLD population,mice and cell models than the control group.2.VEGF-B mainly participates in the pathogenesis of non-alcoholic fatty liver disease by regulating lipogenesis and lipid transport,endoplasmic reticulum stress and apoptosis.Aims 1.To explore the expression of VEGF-B in peripheral blood mononuclear cells(PBMC)of subjects with NAFLD.2.To explore the epigenetic regulation mechanism of VEGF-B in PBMC of subjects with NAFLD.Methods 1.Recruiting NAFLD and health controls in the physical examination center of our hospital.Collecting subjects' blood and separate PBMC.qPCR and WB were used to detect the expression of VEGF-B in PBMC.2.Detecting DNA methylation and histone modification changes in the promoter region of VEGF-B gene in PBMC.Results 1.The expression of VEGF-B in PBMC of NAFLD population was higher than that of health controls.2.DNA methylation and histone modification H3K4me3,H3K9me2,H3K14 ac,H3K18ac in the promoter region of VEGF-B gene in PBMC have no difference between the NAFLD group and the control group,and histone modification H3K27me3 were lower in subjects with NAFLD than the controls.Conclusions 1.The expression of VEGF-B in PBMC of NAFLD subjects was higher than that of controls.2.The expression of VEGF-B may be mediated,at least in part,by epigenetic modifications on VEGF-B promoter histone modification of H3K27me3. |
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