The Expression Of PRDM16 And CIDE-B Protein In Adipose Tissue And Its Relationship With Obesity

Objective To investigate the relationship between the relative expression level of protein PRDM16,CIDE-B and obesity,the protein PRDM16,CIDE-B in subcutaneous adipose and abdominal omental adipose tissues in normal weight and overweight obesity people were determined.And the relationship between the methylation level of PRDM16,CIDE-B gene promoter and obesity was also explored additionally in order to find the relationship between gene methylation and protein expression.Methods126 patients with 25 males and 101 females who accepted the abdominal operation were chosen in this study,patients with pregnant,physical disabilities,mentally disturbed,secondary obesity,malignant tumor and infectious diseases such as hepatitis B,AIDS,tuberculosis were excluded.The data of physiological and biochemical indexes including height,weight,age and blood glucose level were collected.We divided the subjects into normal weight group?18.5kg/??BMI<24kg/??and overweight or obesity group?BMI?24kg/ ??according to the diagnostic criteria for obesity in China.The total protein of subcutaneous adipose tissue and omental adipose tissue was extracted respectively,and the technology western blotting was applied in the following step 8% separation gel,5% concentrated gel for protein PRDM16 and 12%separation gel,5% concentrated gel for protein CIDE-B,and trarsmembrane was conducted after electrophoresis by 2 hours,constant voltage was set at 80 V.Finally the immune response of antibody and secondary antibody was conducted.Put ECL solution configured before on PVDF membrane in gel imaging system,and preserved the pictures of objective band.The gray value of objective band was measured by Gel pro Analyzer software,the relative expression of protein was expressed by the ratio of gray value of objective band and the corresponding gray value of GAPDH.Statistical analysis was conducted by IBM SPSS Statistics 21.0,the difference between two groups was conducted by two independent samples t-test or Mann-Whitney U test,and the difference among three or more groups was conducted by variance analysis or Kruskal-Wallis test,categorical data was analyzed by Chisquare test.The influence of physiological and biochemical factors on the relative expression was explored by multiple linear regression;The difference between subcutaneous and omental adipose tissue of one subject was conducted by Paired sample t test or Wilcoxon Signed rank sum test.the test level ?=0.05?Results1.The result of paired sample t showed the relative expression level of protein PRDM16 in subcutaneous adipose tissue is higher than abdominal omental adipose tissue,but the difference was not statistical significant?t=1.001,P=0.319?;the relative expression level of protein CIDE-B in subcutaneous adipose tissue is higher than abdominal omental adipose tissue,and the difference was statistical significant?t=2.340,P=0.021?.2.In subcutaneous adipose tissue,the difference of relative expression of PRDM16 between normal weight group and overweight group was statistical significant?t=2.587,P=0.011?,and the relative expression of normal weight group is higher than overweight group;In omental adipose tissue,the difference between normal weight group and overweight group was not statistical significant?t=1.671,P=0.097?.3.In subcutaneous adipose tissue,the difference of relative expression of CIDE-B between normal weight group and overweight group was not statistical significant?t=1.558,P=0.122?;In omental adipose tissue,the difference between normal weight group and overweight group was statistical significant?t=-2.253,P=0.026?,and the relative expression of CIDE-B in overweight group is higher than normal weight group.4.In subcutaneous adipose tissue,the relative expression level of CIDE-B with SBP<140mm Hg was lower than that with SBP?140mm Hg,and the difference was statistical significant?t=2.319,P=0.025?.In omental adipose tissue,the relative expression level of CIDE-B with DBP<90mm Hg is less than that with DBP?90mm Hg,and the difference was statistical significant?t=2.290,P=0.024?;In addition,the differences among three TC levels of CIDE-B were statistical significant?F=7.160,P=0.028?,and multiple comparisons showed that the relative expression level of 3.0<TC<5.2mmol/L was less than TC?5.2mmol/L,and the difference was statistical significant?P=0.026?.5.The result of multiple linear regression analysis showed that BMI had correlation with the relative expression of PRDM16 in subcutaneous adipose tissue??=-0.282,P=0.004?,and the correlation was statistical significant,the PRDM16 expression with a lower BMI was higher than that with a higher BMI;In addition,BMI had correlation with the relative expression of PRDM16 in subcutaneous adipose tissue??=0.213,P=0.025?,and the correlation was statistical significant,the CIDE-B expression with a higher BMI was higher than that with a lower BMI.6.The results of correlation analyses showed that the relative expression level of PRDM16 in subcutaneous adipose tissue had positive correlation with that in omental adipose tissue?r=0.482,P<0.001?,and the relative expression level of CIDE-B in subcutaneous adipose tissue had positive correlation with that in omental adipose tissue?r=0.363,P<0.001?.However,There was no linear relationship between PRDM16 and CIDE-B?P>0.05?.7.In subcutaneous adipose tissue,the methylation level of CPG₂8 site in PRDM16 gene had statistical relationship with its relative protein expression??=-3.762,P=0.032?.In omental adipose tissue,the methylation level of CPG₂8 site??=-4.679,P=0.004?in PRDM16 gene had statistical relationship with its relative expression.Conclusions The relationship between the expression of PRDM16 and CIDE-B and obesity in adults may exist.In subcutaneous adipose tissue,the expression of PRDM16 in normal weight group is higher than that in overweight obesity group;In omental adipose tissue,the expression of CIDE-B in normal weight group is lower than that in overweight obesity group;The expression of CIDE-B may have correlation with SBP,DBP and TC level;In addition,the methylation level of CPG₂8 site in PRDM16 gene promoter has correlation with the expression of protein,and high methylation level may depress the expression of protein.