| Objective To analyze the relationship between obesity with diabetes and urinary bisphenol A(BPA),adipose tissue ceramide(Cer)and adipose tissue inflammatory factor levels,and explore the relationship between obesity with diabetes and environmental BPA exposure,and the mediating role of ceramide in population research data for revealing the pathogenesis of metabolic syndrome caused by environmental BPA exposure.Methods From January 2018 to October 2020,200 obese patients who underwent metabolic bariatric surgery in the Gastrointestinal Department of the Second Affiliated Hospital of Anhui Medical University were recruited.The general information and clinical data,urine,blood,and visceral adiposed tissue surgically resected from the patients were collected.Fasting plasma glucose(FPG)and previous medical history divided the subjects into obese patients with diabetes(OD)and obese non-diabetic(OND)patients.Urinary BPA concentration was detected by liquid chromatography-mass spectrometer(LC-MS),and urinary BPA level(BPACr)was corrected by urinary creatinine(Cr).The concentration of Cer in adipose tissue was determined by liquid chromatography-tandem mass spectrometry(LC-MS/MS).The m RNA expressions of ceramide de novo synthetase SPTLC1 and SPTLC2 in adipose tissue of obese patients were detected by reverse transcription-polymerase chain reaction(RT-PCR).Enzyme linked immunosorbent assay(ELISA)was used to measure the levels of inflammatory factors in serum and adipose tissue of obese patients.To investigate the correlation between BPA exposure and inflammatory cytokines,Cers concentrations in adipose tissue,and m RNA expression of de novo synthases SPTLC1 and SPTLC2,we compared the OD and OND urinary BPA levels,serum and adipose tissue inflammatory cytokine levels,adipose tissue concentrations of Cers and synthetases m RNA expression in two groups.Logistic regression was used to analyze the correlation between diabetes and BPA exposure in obese patients.Results1.General information of subjectsAccording to the data of fasting blood glucose and previous medical history,200 obese patients undergoing metabolic bariatric surgery were divided into OD group(n =56)and OND group(n = 144).There were 26 males and 30 females in OD group,with an average age of(34.05±8.21)years,and 48 males and 96 females in OND group,with an average age of(31.43±7.56)years.There were significant differences in age distribution,BMI distribution,TC,TG,FPG and FINS between 2 groups(P<0.05).2.Detection of urinary BPACr,Cer in adipose tissue and inflammatory cytokines in obese patientsThe total detection rate of urinary BPA in 200 obese patients was 96%.The median urinary BPA concentration was 1.80(0.74,3.52)μg/L,and the median urinary BPACR level was 4.30(1.93,7.64)μg/g.The detection rate of Cer in adipose tissue of 200 obese patients was 100%.The content of C18-Cer was 9.35(6.05,12.94)ng/10 mg,the content of C20-Cer was 10.50(4.58,15.45)ng/10 mg,and the content of C24-Cer was 24.63(19.94,29.71)ng/10 mg.The C24:1-Cer content was 40.31(32.81,49.22)ng/10 mg,the C16-Cer content was46.10(39.39,58.37)ng/10 mg,and the total Cer content was 134.37(111.94,156.71)ng/10 mg.The median serum IL-1β concentration of 200 obese patients was 71.77(69.49,74.81)pg/m L,and the median serum IL-18 concentration was 267.52(261.75,278.05)pg/m L.The median IL-1β concentration in adipose tissue was 53.96(48.37,66.07)pg/m L,and the median IL-18 concentration in adipose tissue was 219.02(210.01,252.76)pg/m L.3.Urinary BPACr,adipose tissue Cer and inflammatory factors were correlated with insulin resistance phenotype in obese patientsThe correlation coefficients of urinary BPACr with FPG,FINS and HOMA-IR were 0.301,0.451 and 0.507,respectively(P<0.01).The correlation coefficients of Cer with FPG,FINS and HOMA-IR were 0.310,0.207,0.320,respectively(P<0.01).The correlation coefficients of IL-1β with FPG,FINS and HOMA-IR in adipose tissue were0.425,0.336 and 0.479(P<0.01),respectively.The correlation coefficients between IL-18 and FPG,FINS and HOMA-IR in adipose tissue were 0.353,0.325,0.418(P<0.01),respectively.4.Comparison of BPA exposure levels between obese patients complicated with diabetes and obese non-diabetic patientsThe results of Mann-Whitney rank sum test showed that urinary BPA concentration and urinary BPACr in OD group were higher than those in OND group,with statistical significance(P<0.01),with statistical values of-3.566 and-4.086,respectively.5.Comparison of Cers content in adipose tissue between obese patients complicated with diabetes and obese non-diabetic patientsMann-Whitney rank sum test showed that the levels of 5 types of Cer and total Cer of adipose tissue in OD group were statistically significant compared with those in OND group,except that there was no significant difference in C20-Cer between the other two groups(P<0.05).6.Comparison of SPTLC m RNA expression in adipose tissue between obese patients complicated with diabetes and obese non-diabetic patientsThe results of Mann-Whitney rank sum test showed that the expression of SPTLC1 in OD group was higher than that in OND group(Z=-2.566,P<0.05),and there was no statistical difference in the expression of SPTLC2 between the two groups.7.Comparison of serum and adipose tissue inflammatory factors between obese patients complicated with diabetes and obese non-diabetic patientsThe difference in serum IL-1β between OD group and OND group was statistically significant(P<0.01).The difference in serum IL-18 between OD group and OND group was statistically significant(P<0.01).The difference of IL-1β in adipose tissue between OD group and OND group was statistically significant(P<0.01).The difference of IL-18 in adipose tissue between OD group and OND group was statistically significant(P<0.01).8.Correlation between BPACr and Cer and m RNA expression in adipose tissue in OD and OND groupsThe rank correlation coefficients of urinary BPACr,C16-Cer,C18-Cer,C20-Cer,C24-Cer,C24: 1-Cer and total Cer in OD group were 0.481,0.396,0.165,0.277,0.569 and 0.590,respectively.The rank correlation coefficients between urinary BPACr and C16-Cer,C18-Cer,C20-Cer,C24-Cer,C24:1-Cer and total Cer in OND group were0.405,0.353,0.286,0.183,0.470 and 0.526,respectively.The rank correlation coefficient between BPACr and SPTLC1 expression in OD group was 0.718(P<0.01),and the rank correlation coefficient between BPACr and SPTLC1 expression in OND group was 0.633(P<0.01).There was no statistical correlation between BPACr and SPTLC2 expression in two groups.9.Associations of BPACr with inflammatory cytokines in adipose tissue in OD and OND groupsThe rank correlation coefficients of urinary BPACr and adipose tissue IL-1β and IL-18 in OD group were 0.307 and 0.603,respectively(P<0.01).The rank correlation coefficients between urinary BPACr and adipose tissue IL-1β and IL-18 in OND group were 0.494 and 0.443,respectively(P<0.01).10.Path analysis of the relationship between urinary BPACr and HOMA-IR in obese patientsPath analysis results showed that BPACr had direct and indirect effects on HOMA-IR in obese patients,Cer and inflammatory factors in adipose tissue were the mediator of the association between BPACr and HOMA-IR,and the content of IL-1β in adipose tissue was positively correlated with HOMA-IR.The direct effect of environmental BPA exposure on HOMA-IR in obese patients was 0.328.The effect on the content of IL-1β in adipose tissue by promoting the synthesis of Cer was 0.066 and0.044,respectively.The total effect of BPACr on HOMA-IR was 0.438.11.Logistic regression analysis of BPA exposure and the incidence of diabetes in obese patientsThe incidence of diabetes in obese patients was taken as the dependent variable(non-diabetes =0,diabetes =1),and age,BMI,TC,TG and BPACr were included as independent variables.Multivariate Logistic regression analysis showed that BPACr was one of the risk factors for the development of diabetes in obese patients.Compared with the group with the lowest BPACr level,BPACr was one of the risk factors for diabetes in obese patients.The OR value(95%CI)of BPACr > 7.64μg/g group was 5.44(1.46,20.29).Conclusion1.BPA exposure might be a risk factor for insulin resistance and type 2 diabetes in the population.2.BPA exposure was correlated with ceramide synthase expression and ceramide content in adipose tissue of obese patients.BPA exposure may increase ceramide de novo synthase expression in adipose tissue,leading to the accumulation of ceramide in adipose tissue of obese patients3.BPA exposure promoted insulin resistance in obese patients,which may be related to inflammation of adipose tissue caused by ceramides... |
