Relationship Between Nonalcoholic Fatty Liver Disease And The Single Nucleotide Polymorphism Of Resistin In Type2Diabetes Mellitus Patients

Objective:To investigate the relationship between the resistin gene polymorphism and nonalcoholic fatty liver disease(NAFLD) in type2diabetes mellitus (T2DM) in Han Chinese subjects by analyzed the resistin gene intron+299G/A and5’promoter region-420C/G genotype.Methods:We recruited Han Chinese subjects——738type2diabetes and165control——from the Affiliated Hospital of Binzhou Medical College. The diagnostic standard of T2DM accords with diabetes formulated by WHO(1999). Type1diabetes, secondary diabetes, hyperthyroidism and acute stress state were excluded.All type2diabetes, aged from26to72years,were divided into two groups:395NAFLD, including183males and212females,and343without fatty liver, including162males and181females by ultrasound of liver. The diagnostic standard of NAFLD accords with Clinical Practice Guidelines for NAFLD formulated by_Chinese Association for the Study of Liver Disease[15](2010) and lists as follow:(1)there is no history of alcohol drinking habit or the ethanol intake per week was less than140g in men (70g in women) in the past12months;(2)results of liver imaging study meet the diagnostic criteria of diffuse fatty liver and cannot be explained by other reasons; or patients with metabolic syndrome-related components show a persistent elevation of serum ALT or AST and GGT of unknown causes for more than6months, or both.(3)specific diseases that could lead to steatosis, such as viral hepatitis, drug-induced liver disease, total parenteral nutrition, Wilson’s disease and autoimmune liver disease, can be excluded.The control subjects were composed of126males and153females and aged from26to72years. Endocrinopathy such as diabetes mellitusm, hyperthyroidism,hypothyroidism, Cushing syndrome, acromegaly were excluded. Hypertension, infection, tumour and viral hepatitis were excluded,too. All subjects were measured tall,weight,waist and hip, calculated BMI and waist to hip (WHR). The plasma samples were immediately separated, frozen, and stored at-80℃. Plasma resistin was measured using a human resistin enzyme-linked immunosorbent assay kit following the manufacturer’s protocol as described. The DNA sequence of human resistin was retrievaled in Genebank. The SNP+299G/A and-420C/G genotype was analyzed by polymerase chain reaction-restriction fragment polymorphism. All analyses were performed with SPSS version13.0(SPSS, Chicago, IL). The measurement data of multiple group was compared with single factor analysis of variance,and categorical data compared with Chi-square test. A multivariate non-conditional logistic regression model analysis was used to examine relativity of SNP and T2DM combined with NAFLD. Null hypotheses were rejected at P<0.05.Result:1. Compared with the other two groups, the BMI and WHR are higher in the group of T2DM combined with NAFLD (P<0.05). The TG, TC,non-HDLand LDL are more higher and the HDL is lower in the group of T2DM combined with NAFLD than the controls and the group of not combined with fatty liver in T2DM(P<0.05). The FBG of the group of T2DM combined with NAFLD is higher than the controls(P<0.05).Compared with the other two groups,the FINS and HOMA-IR are higher in the group of T2DM combined with NAFLD(P<0.05). Compared with the other two groups,The ALT,AST and y-GT are more higher in the group of T2DM combined with NAFLD(P<0.05).2. Plasma resistin is the highest in T2DM combined with NAFLD in the three groups(P<0.05). Compared with the controls,the level of Plasma resistin is higher in the group of T2DM not combined with NAFLD(P<0.05). The lever of resistin in-420GG genotype is higher than CC genotype.3. There are CC,CG and GG genotype in resistin gene5’promoter region-420. The frequency of-420GG genotype is lower than other two genotype.The frequency of-420GG genotype in the group of T2DM combined with NAFLD is different from the controls(P<0.01), and different from the group of T2DM not combined with NAFLD(P<0.05),too.The allele frequency of G in the group of T2DM combined with NAFLD is more higher than the group of T2DM not combined with NAFLD and the controls (P<0.05, P<0.01).The risk of incidence of T2DM is higher in-420GG genotype. The risk of incidence of T2DM in-420GG genotype is1.79times to CC genotype,1.59times to CG genotype and1.68times to CC+CG genotype(P<0.01,P<0.05and P<0.05). Resistin-420GG genotype increases the risk of incidence of T2DM.Compared with CC,CG,CC+CG genotype, the risk of incidence of T2DM combined with NAFLD is higher in-420GG genotype(P<0.01,P<0.01and P<0.01). The risk of incidence of T2DM combined with NAFLD in-420GG genotype is2.25times to CC genotype,1.88times to CG genotype and2.05times to CC+CG genotype. Resistin-420GG genotype increases the risk of incidence of T2DM combined with NAFLD.Compared with CC and CC+CG genotype, the risk of incidence of NAFLD in T2DM is higher in-420GG genotype(P<0.05and P<0.05). The risk of incidence of NAFLD in T2DM in-420GG genotype is1.71times to CC genotype and1.60times to CC+CG genotype. Resistin-420GG genotype increases the risk of incidence of NAFLD in T2DM.Compared with CC genotype, the risk of obesity of abdominal type is higher in-420GG genotype(P<0.01). The risk of obesity of abdominal type in-420GG genotype is2.73times to CC genotype. The risk of obesity of abdominal type in the allele G is1.65times to the allele C(P<0.01). The allele G increases the risk of obesity of abdominal type.Compared with CC genotype, the risk of liver dysfunction is higher in-420GG genotype(P<0.01). The risk of liver dysfunction in-420GG genotype is3.97times to CC genotype. The risk of liver dysfunction in the allele G is2.02times to the allele C(P<0.01). The allele G increases the risk of liver dysfunction in T2DM combined with NAFLD.The multivariate non-conditional logistic regression model analysis result showed that-420GG genotype, BMI,GGT,FINS are the risk factors of T2DM combined with NAFLD. The genotype GG increases the risk of incidence of T2DM combined with NAFLD (OR=2.95,95%CI1.15-5.06, P<0.05)4. There are GG,GA and AA genotype in resistin gene intron+299. The frequency of+299AA genotype is lower than other two genotype. The frequency of+299AA genotype in the group of T2DM combined with NAFLD is different from the controls(P<0.05), and different from the group of T2DM not combined with NAFLD(P<0.05),too.The allele frequency of G in the group of T2DM combined with NAFLD is more higher than the group of T2DM not combined with NAFLD and the controls (P<0.05, P<0.01).Compared with GG,GA,GG+GA genotype, the risk of incidence of T2DM is not higher in+299AA genotype.The risk of incidence of T2DM in GG and GA, AA+GA genotype are not different,too. Compared with GG genotype, the risk of incidence of T2DM combined with NAFLD is higher in+299AA and AA+GA genotype(P<0.05, P<0.05). The risk of incidence of T2DM combined with NAFLD in+299AA and AA+GA genotype is1.80and1.45times to GG genotype.In T2DM,the risk of incidence of NAFLD in AA and GG genotype is different (P<0.05). The risk of incidence of NAFLD in AA and GA+GG genotype is different,too (P<0.05). The risk of incidence of NAFLD in+299AA and AA+GA genotype is1.79and1.64times to GG genotype.Compared with GG genotype, the risk of obesity and obesity of abdominal type is higher in+299AA genotype(P<0.01, P<0.01). The risk of obesity and obesity of abdominal type in+299AA genotype is2.05and2.37times to GG genotype. The risk of obesity and obesity of abdominal type in the allele A is1.33and1.44times to the allele G(P<0.01, P<0.01). The multivariate non-conditional logistic regression model analysis result showed that+299AA genotype,BMI,GGT,FINS are the risk factors of T2DM combined with NAFLD. The AA genotype increases the risk of incidence of T2DM combined with NAFLD (OR=2.32,95%CI1.05-4.68, P<0.05)Conclusion:There are CC,CG and GG genotype in resistin gene5’promoter region-420and GG,GA and AA genotype in resistin gene intron+299in Han Chinese subjects.The resistin-420GG genotype increases the risk of incidence of T2DM, increases the risk of incidence of obesity of abdominal type, increases the risk of incidence of NAFLD in T2DM and increases the risk of incidence of T2DM combined with NAFLD. The resistin-420GG genotype related to T2DM combined with NAFLD.The resistin+299AA genotype increases the risk of obesity and obesity of abdominal type, increases the risk of incidence of NAFLD in T2DM and increases the risk of incidence of T2DM combined with NAFLD. The resistin+299AA genotype related to T2DM combined with NAFLD.