2型糖尿病患者抵抗素基因多态性与非酒精性脂肪肝的关系
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摘要
目的：通过检测T2DM患者抵抗素基因5’调控区的-420C/G多态性和内含子区的+299G/A多态性,探讨中国汉族人群中2型糖尿病患者抵抗素基因-420C/G多态性及+299G/A多态性与NAFLD的相关性。
方法：按1999年世界卫生组织制定的糖尿病诊断标准,选取滨州医学院附属医院内分泌科T2DM患者738人,年龄26-72岁,所有2型糖尿病患者均排除1型糖尿病、继发性糖尿病、医源性糖尿病,并排除急性应激状态,如急性心肌梗死、脑梗塞、感染、外伤、手术、糖尿病酮症酸中毒、糖尿病非酮症高渗性昏迷等；甲状腺功能异常；1月内应用影响肾功能的药物,1月内用过降脂药物者。按肝脏超声分为2个亚组：(1)无脂肪肝组343例(男性162例,女性181例),(2)NAFLD组395例(男性183例,女性212例)。NAFLDl临床诊断标准按中华医学会肝脏病学分会脂肪肝和酒精性肝病学组2010年修订的非酒精性脂肪肝病诊疗指南,要求患者无饮酒史或折合乙醇摄入量男性<140g/周,女性<70g/周,并除外药物性肝病、病毒性肝炎、自身免疫性肝病、肝豆状核变性和全胃肠外营养等可致脂肪肝的特定疾病,肝脏彩超表现符合弥漫性脂肪肝的影像学诊断标准,且无其他原因可解释。正常对照组279例(男性126例,女性153例),年龄22-73岁,均为同期滨州医学院附属医院体检中心的健康人群,并除外原发高血压或继发高血压；内分泌系统疾病,如糖耐量减低或糖尿病、库欣综合征、甲状腺功能亢进症或甲状腺功能减退症、肢端肥大症等；急性或慢性感染；肿瘤；病毒性肝炎等,且无饮酒史或折合乙醇摄入量男性<140g/周,女性<70g/周。所有研究对象均清晨空腹,测量身高、体重、腰围、臀围,计算体重指数、腰臀比。采用ELISA法测定血浆抵抗素,采用全自动生化仪测定空腹血糖、谷丙转氨酶、谷草转氨酶、γ-谷氨酰转肽酶、血甘油三酯、血总胆固醇、血高密度脂蛋白及低密度脂蛋白。采用液相平衡竞争放射免疫分析法测定空腹胰岛素。采用TIANGEN公司提供的基因组纯化试剂盒提取基因组DNA。采用聚合酶链反应一限制性片断多态性(PCR-RFLP)方法分析抵抗素420C/G及+299G/A多态性基因型。采用SPSS13.0进行统计学分析。计量资料用mean±SD表示,非正态分布者取自然对数,多样本均数比较采用方差分析,计数资料采用X2检验,以比值比(OR)和95%CI表示相对风险度,相关性分析采用多因素非条件Logistic回归分析。
结果：
1.T2DM合并NAFLD组BMI及WHR的平均值较正常对照组及无脂肪肝组高(P<0.05)。 T2DM合并NAFLD组TG、TC、non-HDL及LDL较正常对照组及无脂肪肝组高(P<0.05),HDL较正常对照组及无脂肪肝组低(P<0.05)。T2DM合并NAFLD组FBG明显高于正常对照组(P<0.05)。T2DM合并NAFLD组的FINS明显较正常对照组及无脂肪肝组高(P<0.05)。T2DM合并NAFLD组的HOMA-IR明显较正常对照组及无脂肪肝组高(P<0.05)。T2DM合并NAFLD组的ALT、 AST及Y-GT较正常对照组及无脂肪肝组高(P<0.05)。
2.T2DM合并NAFLD组抵抗素较正常对照组及无脂肪肝组高(P<0.05)。无脂肪肝组的抵抗素也高于正常对照组(P<0.05)。抵抗素基因-420位点SNP影响血浆抵抗素水平,抵抗素基因-420位点为GG基因型的人群血浆抵抗素水平较高(P<0.05)。
3.正常对照组、T2DM合并NAFLD组及T2DM无脂肪肝组抵抗素基因-420位点均存在CC、CG和GG三种基因型,其中GG基因型频率较低。T2DM合并NAFLD组抵抗素-420C/G基因型分布与正常对照组相比有统计学差异(P<0.01),与无脂肪肝组相比也有统计学差异(P<0.05),G等位基因频率高于正常对照组(P<0.01)及无脂肪肝组(P<0.05)。抵抗素-420位点GG基因型增加了T2DM的风险,是CC基因型的1.79倍(P<0.01),是CG基因型的1.59倍(P<0.05),是C等位基因携带者(CC+CG)的1.68倍(P<0.05)。GG基因型也增加了T2DM合并NAFLD的风险(P<0.01),是CC基因型的2.25倍,是CG基因型的1.88倍,是C等位基因携带者(CC+CG)的2.05倍。抵抗素-420位点为GG基因型的T2DM患者更易并发NAFLD (P<0.05),其发生风险是C等位基因携带者(CC+CG)的1.60倍,是CC基因型的1.71倍。抵抗素-420位点GG基因型发生腹型肥胖的风险高(P<0.01),是CC基因型的2.73倍。抵抗素-420位点为G等位基因的人群发生腹型肥胖的风险升高(P<0.01),是C等位基因的1.65倍。抵抗素-420位点为GG基因型的T2DM合并NAFLD患者更易出现肝功损害(P<0.01),其风险是CC基因型的3.97倍。T2DM合并NAFLD患者中G等位基因携带者发生肝功异常的风险升高(P<0.01),是C等位基因携带者的2.02倍,抵抗素-420位点G等位基因突变增加了T2DM合并NAFLD患者肝功损害发生的风险。多因素非条件Logistc回归分析结果显示：抵抗素-420C/G位点GG基因型与T2MD患者合并NAFLD患病相关,GG基因型的人群,T2DM合并NAFLD患病的风险增加(OR=2.95,95%CI1.15-5.06, P<0.05)。
4.正常对照组、T2DM合并NAFLD组及T2DM无脂肪肝组抵抗素基因+299位点均存在GG、GA和AA三种基因型,其中AA基因型频率较低。T2DM合并NAFLD组抵抗素+299G/A基因型分布与正常对照组相比有统计学差异(P<0.05),与无脂肪肝组相比也有统计学差异(P<0.05),A等位基因频率高于正常对照组(P<0.01)及无脂肪肝组(P<0.05)。抵抗素基因+299位点AA及AA+GA基因型与GG基因型相比T2DM合并NAFLD的风险均有统计学差异(P<0.05), AA、 AA+GA基因型的人群发生T2DM合并NAFLD的风险比GG基因型高,分别是GG基因型的1.80倍及1.45倍。在T2DM人群中,抵抗素+299AA基因型者更易出现NAFLD,与GG及GA+GG基因型相比发生NAFLD的风险均有统计学差异(P<0.05),是GG基因型的1.79倍,是GA+GG基因型的1.64倍。AA基因型比GG基因型更易出现肥胖及腹型肥胖(P值均<0.01),其发生肥胖的风险是GG基因型的2.05倍,发生腹型肥胖的风险是GG基因型的2.37倍。在肝功正常组与肝功异常组两组之间相比抵抗素+299G/A位点基因型分布无统计学差异。多因素非条件Logistc回归分析结果显示：抵抗素+299G/A位点基因型作为独立变量与T2MD患者合并NAFLD患病相关,其中AA基因型是T2DM合并NAFLD患病的危险因素,+299G/A位点AA基因型使T2MD患者合并NAFLD患病的风险增加(OR=2.32,95%CI1.05-4.68, P<0.05)。
结论：
中国汉族人群中抵抗素基因-420位点存在CC、CG和GG三种基因型,+299位点存在GG、GA和AA三种基因型。抵抗素基因-420位点GG基因型升高了血浆抵抗素水平,增加了T2DM的发病风险,增加了腹型肥胖的风险,也增加T2DM并发NAFLD的风险,2型糖尿病人群中抵抗素-420位点为GG基因型的人更易患NAFLD,而如果T2DM并发NAFLD患者的抵抗素基因-420位点为GG基因型则更易出现肝功损害。抵抗素-420位点GG基因型与T2MD患者合并NAFLD患病相关。
抵抗素基因+299位点AA基因型增加了肥胖和腹型肥胖的风险,也增加了T2DM并发NAFLD的风险。T2DM患者如抵抗素基因+299位点为AA基因型则发生NAFLD的风险升高。抵抗素+299位点AA基因型与T2MD患者合并NAFLD患病相关。
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.
方法：按1999年世界卫生组织制定的糖尿病诊断标准,选取滨州医学院附属医院内分泌科T2DM患者738人,年龄26-72岁,所有2型糖尿病患者均排除1型糖尿病、继发性糖尿病、医源性糖尿病,并排除急性应激状态,如急性心肌梗死、脑梗塞、感染、外伤、手术、糖尿病酮症酸中毒、糖尿病非酮症高渗性昏迷等；甲状腺功能异常；1月内应用影响肾功能的药物,1月内用过降脂药物者。按肝脏超声分为2个亚组：(1)无脂肪肝组343例(男性162例,女性181例),(2)NAFLD组395例(男性183例,女性212例)。NAFLDl临床诊断标准按中华医学会肝脏病学分会脂肪肝和酒精性肝病学组2010年修订的非酒精性脂肪肝病诊疗指南,要求患者无饮酒史或折合乙醇摄入量男性<140g/周,女性<70g/周,并除外药物性肝病、病毒性肝炎、自身免疫性肝病、肝豆状核变性和全胃肠外营养等可致脂肪肝的特定疾病,肝脏彩超表现符合弥漫性脂肪肝的影像学诊断标准,且无其他原因可解释。正常对照组279例(男性126例,女性153例),年龄22-73岁,均为同期滨州医学院附属医院体检中心的健康人群,并除外原发高血压或继发高血压；内分泌系统疾病,如糖耐量减低或糖尿病、库欣综合征、甲状腺功能亢进症或甲状腺功能减退症、肢端肥大症等；急性或慢性感染；肿瘤；病毒性肝炎等,且无饮酒史或折合乙醇摄入量男性<140g/周,女性<70g/周。所有研究对象均清晨空腹,测量身高、体重、腰围、臀围,计算体重指数、腰臀比。采用ELISA法测定血浆抵抗素,采用全自动生化仪测定空腹血糖、谷丙转氨酶、谷草转氨酶、γ-谷氨酰转肽酶、血甘油三酯、血总胆固醇、血高密度脂蛋白及低密度脂蛋白。采用液相平衡竞争放射免疫分析法测定空腹胰岛素。采用TIANGEN公司提供的基因组纯化试剂盒提取基因组DNA。采用聚合酶链反应一限制性片断多态性(PCR-RFLP)方法分析抵抗素420C/G及+299G/A多态性基因型。采用SPSS13.0进行统计学分析。计量资料用mean±SD表示,非正态分布者取自然对数,多样本均数比较采用方差分析,计数资料采用X2检验,以比值比(OR)和95%CI表示相对风险度,相关性分析采用多因素非条件Logistic回归分析。
结果：
1.T2DM合并NAFLD组BMI及WHR的平均值较正常对照组及无脂肪肝组高(P<0.05)。 T2DM合并NAFLD组TG、TC、non-HDL及LDL较正常对照组及无脂肪肝组高(P<0.05),HDL较正常对照组及无脂肪肝组低(P<0.05)。T2DM合并NAFLD组FBG明显高于正常对照组(P<0.05)。T2DM合并NAFLD组的FINS明显较正常对照组及无脂肪肝组高(P<0.05)。T2DM合并NAFLD组的HOMA-IR明显较正常对照组及无脂肪肝组高(P<0.05)。T2DM合并NAFLD组的ALT、 AST及Y-GT较正常对照组及无脂肪肝组高(P<0.05)。
2.T2DM合并NAFLD组抵抗素较正常对照组及无脂肪肝组高(P<0.05)。无脂肪肝组的抵抗素也高于正常对照组(P<0.05)。抵抗素基因-420位点SNP影响血浆抵抗素水平,抵抗素基因-420位点为GG基因型的人群血浆抵抗素水平较高(P<0.05)。
3.正常对照组、T2DM合并NAFLD组及T2DM无脂肪肝组抵抗素基因-420位点均存在CC、CG和GG三种基因型,其中GG基因型频率较低。T2DM合并NAFLD组抵抗素-420C/G基因型分布与正常对照组相比有统计学差异(P<0.01),与无脂肪肝组相比也有统计学差异(P<0.05),G等位基因频率高于正常对照组(P<0.01)及无脂肪肝组(P<0.05)。抵抗素-420位点GG基因型增加了T2DM的风险,是CC基因型的1.79倍(P<0.01),是CG基因型的1.59倍(P<0.05),是C等位基因携带者(CC+CG)的1.68倍(P<0.05)。GG基因型也增加了T2DM合并NAFLD的风险(P<0.01),是CC基因型的2.25倍,是CG基因型的1.88倍,是C等位基因携带者(CC+CG)的2.05倍。抵抗素-420位点为GG基因型的T2DM患者更易并发NAFLD (P<0.05),其发生风险是C等位基因携带者(CC+CG)的1.60倍,是CC基因型的1.71倍。抵抗素-420位点GG基因型发生腹型肥胖的风险高(P<0.01),是CC基因型的2.73倍。抵抗素-420位点为G等位基因的人群发生腹型肥胖的风险升高(P<0.01),是C等位基因的1.65倍。抵抗素-420位点为GG基因型的T2DM合并NAFLD患者更易出现肝功损害(P<0.01),其风险是CC基因型的3.97倍。T2DM合并NAFLD患者中G等位基因携带者发生肝功异常的风险升高(P<0.01),是C等位基因携带者的2.02倍,抵抗素-420位点G等位基因突变增加了T2DM合并NAFLD患者肝功损害发生的风险。多因素非条件Logistc回归分析结果显示：抵抗素-420C/G位点GG基因型与T2MD患者合并NAFLD患病相关,GG基因型的人群,T2DM合并NAFLD患病的风险增加(OR=2.95,95%CI1.15-5.06, P<0.05)。
4.正常对照组、T2DM合并NAFLD组及T2DM无脂肪肝组抵抗素基因+299位点均存在GG、GA和AA三种基因型,其中AA基因型频率较低。T2DM合并NAFLD组抵抗素+299G/A基因型分布与正常对照组相比有统计学差异(P<0.05),与无脂肪肝组相比也有统计学差异(P<0.05),A等位基因频率高于正常对照组(P<0.01)及无脂肪肝组(P<0.05)。抵抗素基因+299位点AA及AA+GA基因型与GG基因型相比T2DM合并NAFLD的风险均有统计学差异(P<0.05), AA、 AA+GA基因型的人群发生T2DM合并NAFLD的风险比GG基因型高,分别是GG基因型的1.80倍及1.45倍。在T2DM人群中,抵抗素+299AA基因型者更易出现NAFLD,与GG及GA+GG基因型相比发生NAFLD的风险均有统计学差异(P<0.05),是GG基因型的1.79倍,是GA+GG基因型的1.64倍。AA基因型比GG基因型更易出现肥胖及腹型肥胖(P值均<0.01),其发生肥胖的风险是GG基因型的2.05倍,发生腹型肥胖的风险是GG基因型的2.37倍。在肝功正常组与肝功异常组两组之间相比抵抗素+299G/A位点基因型分布无统计学差异。多因素非条件Logistc回归分析结果显示：抵抗素+299G/A位点基因型作为独立变量与T2MD患者合并NAFLD患病相关,其中AA基因型是T2DM合并NAFLD患病的危险因素,+299G/A位点AA基因型使T2MD患者合并NAFLD患病的风险增加(OR=2.32,95%CI1.05-4.68, P<0.05)。
结论：
中国汉族人群中抵抗素基因-420位点存在CC、CG和GG三种基因型,+299位点存在GG、GA和AA三种基因型。抵抗素基因-420位点GG基因型升高了血浆抵抗素水平,增加了T2DM的发病风险,增加了腹型肥胖的风险,也增加T2DM并发NAFLD的风险,2型糖尿病人群中抵抗素-420位点为GG基因型的人更易患NAFLD,而如果T2DM并发NAFLD患者的抵抗素基因-420位点为GG基因型则更易出现肝功损害。抵抗素-420位点GG基因型与T2MD患者合并NAFLD患病相关。
抵抗素基因+299位点AA基因型增加了肥胖和腹型肥胖的风险,也增加了T2DM并发NAFLD的风险。T2DM患者如抵抗素基因+299位点为AA基因型则发生NAFLD的风险升高。抵抗素+299位点AA基因型与T2MD患者合并NAFLD患病相关。
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.
引文
1. Ruhl CE,Everhart JE.Epidemiology of nonalcoholic fatty liver.Clin Liver Dis, 2004,8(3):501-519.
2. Fan JG, Farrell GC.Epidemiology of non-alcoholic fatty liver disease in China. J Hepatol,2009,50(1):204-210.
3. Day CP.Steatohepatitis:a tale of two "hits"?Gustroenteroligy,1998,114(4):824-825.
4. Zhang Y, Proenca R,Maffei M,et al.Positional cloning of the mouse obese gene and its human homologue.Nature,1994,372(6505):425-432.
5. Uygun A,Kadayifci A,Yesilova Z,et al.Serum leptin levels in patients with nonalcoholic steatohepatitis. Am J Gastroenterol,2000,95(12):3584-3589.
6. Tobe K, Suzuki R, Aoyama M,et al.Increased expression of the sterol regulatory element-binding protein-1 gene in insulin receptor substrate-2(-/-)mouse liver.J Biol Chem,2001,276(42):3837-3840.
7. Kim KO,Park SH,Han TH,et al.Relationship between the severity of liver damage and the serum leptin level for nonalcoholic fatty liver disease.Korean J Hepatol,2005,11(1):51-58.
8. Yamauchi T,Kamon J,Ito Y,et al.Cloning of adiponectin receptors that mediate antidiabetic metabolic effects. Nature,2003,423(6941):762-769.
9. Anania FA.Adiponectin and alcoholic fatty liver:is it, after all, about what you eat? Hepatology,2005,42(3):530-532.
10. You M, Considine R, Leone T,et al. Role of adiponectin in the protective action of dietary saturated fat against alcoholic fatty liver in mice.Hepatology, 2005,42(3):568-577.
11. Shklyaev S, Aslanidi G, Tennant M, et al.Sustained peripheral expression of transgene adiponectin offsets the development of diet-induced obesity in rats. Proc Natl Acad Sci U S A,2003,100(24):14217-14222.
12. Lopez-Bermejo A, Botas P, Funahashi T,et al. Adiponectin, hepatocellular dysfunction and insulin sensitivity. Clin Endocrinol (Oxf),2004,60(2):256- 263.
13. Targher G, Bertolini L, Scala L,et al. Decreased plasma adiponectin concentrations are closely associated with nonalcoholic hepatic steatosis in obese individuals. Clin Endocrinol (Oxf),2004,61 (6):700-703.
14. Hui JM, Hodge A, Farrell GC,et al. Beyond insulin resistance in NASH: TNF-alpha or adiponectin? Hepatology,2004,40(1):46-54.
15. Xu A,Wang Y,Keshaw H,et al.The fat-derived hormone adiponectin alleviares alcoholic and nonalcoholic fatty liver diseases in mice.J Clin Invest,2003,112(1):91-100.
16. Miura A, Ishizuka T, Kanoh Y,et al. Effect of tumor necrosis factor-alpha on insulin signal transduction in rat adipocytes:relation to PKCbeta and zeta translocation. Biochim Biophys Acta,1999,1449(3):227-238.
17. Brindley DN, Wang CN, Mei J,et al.Tumor necrosis factor-alpha and ceramides in insulin resistance. Lipids,1999,34 (Suppl):S85-S88.
18. Lewis G F, Carpentier A, Adeli K. Disordered fat storage and mobilization in the pathogenesis of insulin resistance and type 2 diabetes.Endocr Rev,2002,23(2):201-229.
19. Feldstein AE, Werneburg NW, Canbay A,et al. Free fatty acids promote hepatic lipotoxicity by stimulating TNF-alpha expression via a lysosomal pathway. Hepatology,2004,40(1):185-194.
20. Alwayn IP, Andersson C, Lee S, et al.Inhibition of matrix metallop roteinases increases PPAR-alpha and IL-6 and prevents dietary-induced hepatic steatosis and injury in a murine model.Am J Physiol Gastrointest Liver Physiol,2006,291(6):G1011-G1019.
21. Lutchman G, Promrat K, Kleiner D E, et al.Changes in serum adipokine levels during pioglitazone treatment for nonalcoholic steatohepatitis:relationship to histological imp rovement. Clin Gastroenterol Hepatol,2006,4 (8):1048-1052.
22. Everett L, Galii A, Crabb D. The role of hepatic peroxisome proliferator-activated receptors (PPARs) in health and disease. Liver,2000, 20(3):191-199.
23. Steppan CM,Bailey ST,Bhat S, et al. The hormone resistin links obesity to diabetes. Nature,2001,409(1):307-312.
24. Steppan CM,Brown EJ,Wright CM,et al. A family of tissue-specific resistin-like molecules. Proc Natl Acad Sci U S A,2001,98(2):502-506.
25. Banerjee RR,Lazar MA. Dimerization of resistin and resistin-like molecules is determined by a single cysteine. J Biol Chem,2001,276(28):25970-25973.
26. Kim KH,Lee K,Moon YS, et al. A cysteine-rich adipose tissue-specific secretory factor inhibits adipocyte differentiation. J Biol Chem,2001, 276(14):11252-11256.
27. Nagaev I,Smith U. Insulin resistance and type 2 diabetes are not related to resistin expression in human fat cells or skeletal muscle. Biochem Biophys Res commun,2001,285(2):561-564.
28. Savage DB,Sewter CP,Klenk ES,et al. Resistin/Fizz3 expression in relation to obesity and peroxisome proliferator-activated receptor-gamme action in humans. Diabetes,2001,50(10):2199-2202.
29. Janke J,Engeli S,Gorzelniak K, et al. Resistin gene expression in human adipocytes is not related to insulin resistance. Obes Res,2002,10(9300):1-5.
30. Mcternan CL,Mcternan PG,Harte AL, et al. Resistin,central obesity,and type 2 diabetes. Lancet,2002,359(9300):46-47.
31. Mcternan PG,Mcternan CL,Chetty R, et al. Increased resistin gene and protien in human abdominal adipose tissue. J Clin Endocrinol Metab,2002, 87(5):2407.
32. Silswal N,Singh AK,Aruna B,et al.Human resistin stimulates the pro-inflammalory cytokines TNF-alpha and IL-12 in macrophages by NF-kappaB-dependent parhway. Biochem Biophys Res Commun,2005,334(4): 1092-1101.
33. Bokarewa M,Nagaev I,Dahlberg L, et al. Resistin, an adipokine with potent proinflammatory properties.J immunol,2005,174(9):5789-5795.
34. Lee J H,Chan J L,Yiannakouris N,et al.Circulating resistin levels are not associated with obesity or insulin resistance in humans and are not regulated by fasting or leptin administration:cross-sectional and interventional studies in normal,insulin-resistant,and diabetic subjects.J Clin Endocrinol Metab,2003,88(10):4848-4856.
35. Kralisch S,Sommer G,Stangl V,et al.Secretory products from human adipocytes impair endothelial function via nuclear factor kappaB.Atherosclerosis,2008,196(2):523-531.
36. Muse ED,Obici S,Bhanot S,et al.Role of resistin in diet-in-duced hepatic insulin resistance.J Clin Ivest,2004,114(2):232-239.
37. Bajaj M, Suraamornkul S, Hardies LJ, et al. Plasma resistin concentration,hepitic fat content,and hepatic and peripheral insulin resistance in pioglitazone treated type II diabetic patients.J Obes Relat Matab Disord,2004, 28(6):783-789.
38. Song H, Shojima N, Sakoda H, et al. Resistin is regulated by C/EBPa and PPARy, and signal-transducing molecules. J Bio-chem Biophys Res Commun, 2002,299(2):291-298.
39. Zhou L, Li Y, Xia T, et al.Resistin overexpression impaired glucose tolerance in hepatocytes. Eur Cytokine Netw,2006,17(3):189-195.
40.范建高,方继伟,陆元善,等.高脂饮食非酒精性脂肪性肝病大鼠肝脏成脂相关基因表达增强.中华消化杂志,2006,26(5)：320-324.
41. Fu Y, Luo L, Luo N, et al.Proinflammatory cytokine production and insulin sensitivity regulated by over expression of resistin in 3T3-L1 adipocytes.Nutr Metab,2006,3:28.
42. Jiang LL,Li L,Hong XF,et al.Patients with nonalcoholic fatty liver disease increased serum resistin levers and decreased adiponectin levels.Eur J Gastroenterol,2009,21(6):662-666.
43. Lebensztejn DM,Wojtkowaka M,Skiba E,et al.Serum concentration of adiponectin,leptin and resistin in obese chindren with non-alcoholic fatty liver disease.Adv Med Sci,2009,54(2):177-182.
44. Sentinelli F,Romeo S,Arca M,et al.Human resistin gene,obesity,and type 2 diabetes mutation analysis and population study.Diabetes,2002,51(3):860-862.
45. Ma X,Warram JB,Trischitta V,et al.Genetic variants at the resistin locus and risk of type 2 diabetes in Caucasians. J Clin Endocrinol Metab,2002,87(9):4407-4410.
46. Engert JC,Vohl MC,Williams SM,et al.5'flanking varians of resistin are associated with obesity. Diabetes,2002,51(5):1629-1634.
47. Osawa H,Onuma H,Murakami A,et al.Systematic search for single nucleotide polymorphisms in the resistin gene:the absence of evidence for the association of three identified single nucleotide polymorphisms with Japanese type 2 diabetes.Diabetes,2002,51(5):863-866.
48.梁梅花,张中超,陈峰,等.中国汉族人群resistin基因5'调控区与2型糖尿病的关系.哈尔滨医科大学学报,2003,37(4)：321-323.
49.董艳,李果,骆天红,等.抵抗素基因多态性与华东地区2型糖尿病的相关性.上海第二医科大学学报,2003,23(2)：97-99.
50. Baba T, Endo T, Sata F,et al.The contributions of resistin and adiponectin gene single nucleotide polymorphisms to the genetic risk for polycystic ovary syndrome in a Japanese population. Gynecol Endocrinol.2009,25(8):498-503.
51. Tsukahara T, Nakashima E, Watarai A,et al. Polymorphism in resistin promoter region at-420 determines the serum resistin levels and may be a risk marker of stroke in Japanese type 2 diabetic patients. Diabetes Res Clin Pract. 2009,84(2):179-186.
52.张维,毕娅欣.抵抗素基因多态性与2型糖尿病及肥胖相关性研究.实用全科医学,2007,5(6)：478-479.
53.董惠萍,闫胜利,赵世华,等.抵抗素基因299G/A多态性与2型糖尿病的相关性研究.中华糖尿病杂志,2004,12(6)：431-432.
54.邹彩艳,李茵茵,张红霞,等.抵抗素基因+299G/A多态性与T2DM并高血压病的相关性.山东医药,2007,47(10)：3-4.
55. Tan MS, Chang SY, Chang DM,et al. Association of resistin gene 3'-untranslated region +62G-->A polymorphism with type 2 diabetes and hypertension in a Chinese population. J Clin Endocrinol Metab.2003,88(3):1258-1263.
56.柳亢宗,林梅,黄浩,等.抵抗素基因外显子4第1308G/A多态性与2型糖尿病相关性研究.临床内科杂志,2005,22(9)：619-621.
57. McCullough AJ. Pathophysiology of nonalcoholic steatohepatitis. J Clin Gastroenterol,2006,40 (Suppl 1):S17-S29.
58. Sreekumar R, Rosado B, Rasmussen D, et al. Hepatic gene expression in histologically progressive nonalcoholic steatohepatitis. Hepatology,2003,38(1): 244-251.
59.石胜利,聂玉强,李喻元.脂联素基因Gly15Gly多态性与非酒精性脂肪肝病的关系.广州医学院学报,2005(6),33：1-3.
60. Tokushige K, Hashimoto E, Noto H,et al.Influence of adiponectin gene polymorphisms in Japanese patients with non-alcoholic fatty liver disease. J Gastroenterol.2009;44(9):976-982.
61.丁百静,王震,杨勇,等.2型糖尿病患者LEP-2548位点SNP与非酒精性脂肪肝的关系.世界华人消化杂志,2009,17(26)：2732-2737。
62.赖晓波,聂玉强,李瑜元,等.非酒精性脂肪肝患者瘦素基因G2548A位点多态性与胰岛素抵抗的关系-附107例报告.新医学,2008,39(12)：781-782.
63. Valenti L, FracanzaniAL, Dongiovani P, et al. Tumor necrosis factor alpha promoter polymorphisms and insulin resistance in nonalcoholic fatty liver disease. Gastroenterology,2002,122(2):274-280.
64.黄健,李瑜元.非酒精性脂肪肝患者肿瘤坏死因子2α基因多态性研究.广州医学院学报,2004,32(4)：24-26.
65.马瑞宏,黄颖秋,西元利治.抵抗素基因启动子-420C/G多态性与日本高知地区非酒精性脂肪肝炎的相关性.世界华人消化杂志,2007,15(30)：3204-3209.
66.中华医学会肝脏病学分会脂肪肝和酒精性肝病学组.非酒精性脂肪肝病诊疗指南.胃肠病学,2010,15(11)：676-680.
67. Farrell GC, Larter CZ.Nonalcoholic fatty liver disease:from steatosis to cirrhosis. Hepatology,2006,43(2 suppl 1):S99-S112.
68. de Alwis NM,Day CP. Nonalcoholic fatty liver disease:the mist gradually clears.J Hepatol,2008,48(suppl 1):S104-S112.
69. Angulo P.GI epideminology:nonalcoholic fatty liver disease.Aliment Pharmcol Ther,2007,25(8):883-889.
70. Fan JQSaibara T,Chitturi S,et al.What are the risk factors and settings for non-alcoholic fatty liver disease in Asia-Pacific?J Gastroenterol Hepatol,2007,22(6):794-800.
71. Yang RZ,Huang Q,Xu A,et al.Comparative studies of resistin expression and phylogenomica in human and mouse.Biochem Biophys Res Commun, 2003,310(3):927-935.
72. Wiegand J,Mossner J,Tillmann HL. Non-alcoholic fatty liver disease and non-alcoholic steatohepatitis.Internist(Berl),2007,48(2):154-163.
73. Chitturi S,Abeygunasekera S,Farrell GC, et al. NASH and insulin resistance:Insulin hypersecretion and specific association with the insulin resistance syndrome.Hepatology,2002,35(2):373-379.
74. Farese RV,Sajan MP,Standaert ML.Insulin-sensitive protein kinases(atypical protein kinase C and protein kinase B/Akt):actions and defects in obesity and type Ⅱ diabetes.Exp Biol Med(Maywood),2005,230(9):593-605.
75. Tilg H,Diehl AM.Cytokines in alcoholic and nonalcoholic steatohepatitis.N Engl J Med,2000,343(20):1467-1476.
76. Vernochet C,Peres SB,Davis KE,et al.C/EBPalpha and the corepressors CtBP1 and CtBP2 regulate repression of select visceral white adipose genes during induction of the brown phenotype in white adipocytes by peroxisome proliferator-activated receptor gamma agonists. Mol Cell Biol, 2009,29(17):4714-4728.
77. Tomaru T, Steger DJ, Lefterova MI, et al. Adipocyte-specific expression of murine resistin is mediated by synergism between peroxisome proliferator-activated receptor gamma and CCAAT/enhancer-binding proteins. J Biol Chem,2009,284(10):6116-6125.
2. Fan JG, Farrell GC.Epidemiology of non-alcoholic fatty liver disease in China. J Hepatol,2009,50(1):204-210.
3. Day CP.Steatohepatitis:a tale of two "hits"?Gustroenteroligy,1998,114(4):824-825.
4. Zhang Y, Proenca R,Maffei M,et al.Positional cloning of the mouse obese gene and its human homologue.Nature,1994,372(6505):425-432.
5. Uygun A,Kadayifci A,Yesilova Z,et al.Serum leptin levels in patients with nonalcoholic steatohepatitis. Am J Gastroenterol,2000,95(12):3584-3589.
6. Tobe K, Suzuki R, Aoyama M,et al.Increased expression of the sterol regulatory element-binding protein-1 gene in insulin receptor substrate-2(-/-)mouse liver.J Biol Chem,2001,276(42):3837-3840.
7. Kim KO,Park SH,Han TH,et al.Relationship between the severity of liver damage and the serum leptin level for nonalcoholic fatty liver disease.Korean J Hepatol,2005,11(1):51-58.
8. Yamauchi T,Kamon J,Ito Y,et al.Cloning of adiponectin receptors that mediate antidiabetic metabolic effects. Nature,2003,423(6941):762-769.
9. Anania FA.Adiponectin and alcoholic fatty liver:is it, after all, about what you eat? Hepatology,2005,42(3):530-532.
10. You M, Considine R, Leone T,et al. Role of adiponectin in the protective action of dietary saturated fat against alcoholic fatty liver in mice.Hepatology, 2005,42(3):568-577.
11. Shklyaev S, Aslanidi G, Tennant M, et al.Sustained peripheral expression of transgene adiponectin offsets the development of diet-induced obesity in rats. Proc Natl Acad Sci U S A,2003,100(24):14217-14222.
12. Lopez-Bermejo A, Botas P, Funahashi T,et al. Adiponectin, hepatocellular dysfunction and insulin sensitivity. Clin Endocrinol (Oxf),2004,60(2):256- 263.
13. Targher G, Bertolini L, Scala L,et al. Decreased plasma adiponectin concentrations are closely associated with nonalcoholic hepatic steatosis in obese individuals. Clin Endocrinol (Oxf),2004,61 (6):700-703.
14. Hui JM, Hodge A, Farrell GC,et al. Beyond insulin resistance in NASH: TNF-alpha or adiponectin? Hepatology,2004,40(1):46-54.
15. Xu A,Wang Y,Keshaw H,et al.The fat-derived hormone adiponectin alleviares alcoholic and nonalcoholic fatty liver diseases in mice.J Clin Invest,2003,112(1):91-100.
16. Miura A, Ishizuka T, Kanoh Y,et al. Effect of tumor necrosis factor-alpha on insulin signal transduction in rat adipocytes:relation to PKCbeta and zeta translocation. Biochim Biophys Acta,1999,1449(3):227-238.
17. Brindley DN, Wang CN, Mei J,et al.Tumor necrosis factor-alpha and ceramides in insulin resistance. Lipids,1999,34 (Suppl):S85-S88.
18. Lewis G F, Carpentier A, Adeli K. Disordered fat storage and mobilization in the pathogenesis of insulin resistance and type 2 diabetes.Endocr Rev,2002,23(2):201-229.
19. Feldstein AE, Werneburg NW, Canbay A,et al. Free fatty acids promote hepatic lipotoxicity by stimulating TNF-alpha expression via a lysosomal pathway. Hepatology,2004,40(1):185-194.
20. Alwayn IP, Andersson C, Lee S, et al.Inhibition of matrix metallop roteinases increases PPAR-alpha and IL-6 and prevents dietary-induced hepatic steatosis and injury in a murine model.Am J Physiol Gastrointest Liver Physiol,2006,291(6):G1011-G1019.
21. Lutchman G, Promrat K, Kleiner D E, et al.Changes in serum adipokine levels during pioglitazone treatment for nonalcoholic steatohepatitis:relationship to histological imp rovement. Clin Gastroenterol Hepatol,2006,4 (8):1048-1052.
22. Everett L, Galii A, Crabb D. The role of hepatic peroxisome proliferator-activated receptors (PPARs) in health and disease. Liver,2000, 20(3):191-199.
23. Steppan CM,Bailey ST,Bhat S, et al. The hormone resistin links obesity to diabetes. Nature,2001,409(1):307-312.
24. Steppan CM,Brown EJ,Wright CM,et al. A family of tissue-specific resistin-like molecules. Proc Natl Acad Sci U S A,2001,98(2):502-506.
25. Banerjee RR,Lazar MA. Dimerization of resistin and resistin-like molecules is determined by a single cysteine. J Biol Chem,2001,276(28):25970-25973.
26. Kim KH,Lee K,Moon YS, et al. A cysteine-rich adipose tissue-specific secretory factor inhibits adipocyte differentiation. J Biol Chem,2001, 276(14):11252-11256.
27. Nagaev I,Smith U. Insulin resistance and type 2 diabetes are not related to resistin expression in human fat cells or skeletal muscle. Biochem Biophys Res commun,2001,285(2):561-564.
28. Savage DB,Sewter CP,Klenk ES,et al. Resistin/Fizz3 expression in relation to obesity and peroxisome proliferator-activated receptor-gamme action in humans. Diabetes,2001,50(10):2199-2202.
29. Janke J,Engeli S,Gorzelniak K, et al. Resistin gene expression in human adipocytes is not related to insulin resistance. Obes Res,2002,10(9300):1-5.
30. Mcternan CL,Mcternan PG,Harte AL, et al. Resistin,central obesity,and type 2 diabetes. Lancet,2002,359(9300):46-47.
31. Mcternan PG,Mcternan CL,Chetty R, et al. Increased resistin gene and protien in human abdominal adipose tissue. J Clin Endocrinol Metab,2002, 87(5):2407.
32. Silswal N,Singh AK,Aruna B,et al.Human resistin stimulates the pro-inflammalory cytokines TNF-alpha and IL-12 in macrophages by NF-kappaB-dependent parhway. Biochem Biophys Res Commun,2005,334(4): 1092-1101.
33. Bokarewa M,Nagaev I,Dahlberg L, et al. Resistin, an adipokine with potent proinflammatory properties.J immunol,2005,174(9):5789-5795.
34. Lee J H,Chan J L,Yiannakouris N,et al.Circulating resistin levels are not associated with obesity or insulin resistance in humans and are not regulated by fasting or leptin administration:cross-sectional and interventional studies in normal,insulin-resistant,and diabetic subjects.J Clin Endocrinol Metab,2003,88(10):4848-4856.
35. Kralisch S,Sommer G,Stangl V,et al.Secretory products from human adipocytes impair endothelial function via nuclear factor kappaB.Atherosclerosis,2008,196(2):523-531.
36. Muse ED,Obici S,Bhanot S,et al.Role of resistin in diet-in-duced hepatic insulin resistance.J Clin Ivest,2004,114(2):232-239.
37. Bajaj M, Suraamornkul S, Hardies LJ, et al. Plasma resistin concentration,hepitic fat content,and hepatic and peripheral insulin resistance in pioglitazone treated type II diabetic patients.J Obes Relat Matab Disord,2004, 28(6):783-789.
38. Song H, Shojima N, Sakoda H, et al. Resistin is regulated by C/EBPa and PPARy, and signal-transducing molecules. J Bio-chem Biophys Res Commun, 2002,299(2):291-298.
39. Zhou L, Li Y, Xia T, et al.Resistin overexpression impaired glucose tolerance in hepatocytes. Eur Cytokine Netw,2006,17(3):189-195.
40.范建高,方继伟,陆元善,等.高脂饮食非酒精性脂肪性肝病大鼠肝脏成脂相关基因表达增强.中华消化杂志,2006,26(5)：320-324.
41. Fu Y, Luo L, Luo N, et al.Proinflammatory cytokine production and insulin sensitivity regulated by over expression of resistin in 3T3-L1 adipocytes.Nutr Metab,2006,3:28.
42. Jiang LL,Li L,Hong XF,et al.Patients with nonalcoholic fatty liver disease increased serum resistin levers and decreased adiponectin levels.Eur J Gastroenterol,2009,21(6):662-666.
43. Lebensztejn DM,Wojtkowaka M,Skiba E,et al.Serum concentration of adiponectin,leptin and resistin in obese chindren with non-alcoholic fatty liver disease.Adv Med Sci,2009,54(2):177-182.
44. Sentinelli F,Romeo S,Arca M,et al.Human resistin gene,obesity,and type 2 diabetes mutation analysis and population study.Diabetes,2002,51(3):860-862.
45. Ma X,Warram JB,Trischitta V,et al.Genetic variants at the resistin locus and risk of type 2 diabetes in Caucasians. J Clin Endocrinol Metab,2002,87(9):4407-4410.
46. Engert JC,Vohl MC,Williams SM,et al.5'flanking varians of resistin are associated with obesity. Diabetes,2002,51(5):1629-1634.
47. Osawa H,Onuma H,Murakami A,et al.Systematic search for single nucleotide polymorphisms in the resistin gene:the absence of evidence for the association of three identified single nucleotide polymorphisms with Japanese type 2 diabetes.Diabetes,2002,51(5):863-866.
48.梁梅花,张中超,陈峰,等.中国汉族人群resistin基因5'调控区与2型糖尿病的关系.哈尔滨医科大学学报,2003,37(4)：321-323.
49.董艳,李果,骆天红,等.抵抗素基因多态性与华东地区2型糖尿病的相关性.上海第二医科大学学报,2003,23(2)：97-99.
50. Baba T, Endo T, Sata F,et al.The contributions of resistin and adiponectin gene single nucleotide polymorphisms to the genetic risk for polycystic ovary syndrome in a Japanese population. Gynecol Endocrinol.2009,25(8):498-503.
51. Tsukahara T, Nakashima E, Watarai A,et al. Polymorphism in resistin promoter region at-420 determines the serum resistin levels and may be a risk marker of stroke in Japanese type 2 diabetic patients. Diabetes Res Clin Pract. 2009,84(2):179-186.
52.张维,毕娅欣.抵抗素基因多态性与2型糖尿病及肥胖相关性研究.实用全科医学,2007,5(6)：478-479.
53.董惠萍,闫胜利,赵世华,等.抵抗素基因299G/A多态性与2型糖尿病的相关性研究.中华糖尿病杂志,2004,12(6)：431-432.
54.邹彩艳,李茵茵,张红霞,等.抵抗素基因+299G/A多态性与T2DM并高血压病的相关性.山东医药,2007,47(10)：3-4.
55. Tan MS, Chang SY, Chang DM,et al. Association of resistin gene 3'-untranslated region +62G-->A polymorphism with type 2 diabetes and hypertension in a Chinese population. J Clin Endocrinol Metab.2003,88(3):1258-1263.
56.柳亢宗,林梅,黄浩,等.抵抗素基因外显子4第1308G/A多态性与2型糖尿病相关性研究.临床内科杂志,2005,22(9)：619-621.
57. McCullough AJ. Pathophysiology of nonalcoholic steatohepatitis. J Clin Gastroenterol,2006,40 (Suppl 1):S17-S29.
58. Sreekumar R, Rosado B, Rasmussen D, et al. Hepatic gene expression in histologically progressive nonalcoholic steatohepatitis. Hepatology,2003,38(1): 244-251.
59.石胜利,聂玉强,李喻元.脂联素基因Gly15Gly多态性与非酒精性脂肪肝病的关系.广州医学院学报,2005(6),33：1-3.
60. Tokushige K, Hashimoto E, Noto H,et al.Influence of adiponectin gene polymorphisms in Japanese patients with non-alcoholic fatty liver disease. J Gastroenterol.2009;44(9):976-982.
61.丁百静,王震,杨勇,等.2型糖尿病患者LEP-2548位点SNP与非酒精性脂肪肝的关系.世界华人消化杂志,2009,17(26)：2732-2737。
62.赖晓波,聂玉强,李瑜元,等.非酒精性脂肪肝患者瘦素基因G2548A位点多态性与胰岛素抵抗的关系-附107例报告.新医学,2008,39(12)：781-782.
63. Valenti L, FracanzaniAL, Dongiovani P, et al. Tumor necrosis factor alpha promoter polymorphisms and insulin resistance in nonalcoholic fatty liver disease. Gastroenterology,2002,122(2):274-280.
64.黄健,李瑜元.非酒精性脂肪肝患者肿瘤坏死因子2α基因多态性研究.广州医学院学报,2004,32(4)：24-26.
65.马瑞宏,黄颖秋,西元利治.抵抗素基因启动子-420C/G多态性与日本高知地区非酒精性脂肪肝炎的相关性.世界华人消化杂志,2007,15(30)：3204-3209.
66.中华医学会肝脏病学分会脂肪肝和酒精性肝病学组.非酒精性脂肪肝病诊疗指南.胃肠病学,2010,15(11)：676-680.
67. Farrell GC, Larter CZ.Nonalcoholic fatty liver disease:from steatosis to cirrhosis. Hepatology,2006,43(2 suppl 1):S99-S112.
68. de Alwis NM,Day CP. Nonalcoholic fatty liver disease:the mist gradually clears.J Hepatol,2008,48(suppl 1):S104-S112.
69. Angulo P.GI epideminology:nonalcoholic fatty liver disease.Aliment Pharmcol Ther,2007,25(8):883-889.
70. Fan JQSaibara T,Chitturi S,et al.What are the risk factors and settings for non-alcoholic fatty liver disease in Asia-Pacific?J Gastroenterol Hepatol,2007,22(6):794-800.
71. Yang RZ,Huang Q,Xu A,et al.Comparative studies of resistin expression and phylogenomica in human and mouse.Biochem Biophys Res Commun, 2003,310(3):927-935.
72. Wiegand J,Mossner J,Tillmann HL. Non-alcoholic fatty liver disease and non-alcoholic steatohepatitis.Internist(Berl),2007,48(2):154-163.
73. Chitturi S,Abeygunasekera S,Farrell GC, et al. NASH and insulin resistance:Insulin hypersecretion and specific association with the insulin resistance syndrome.Hepatology,2002,35(2):373-379.
74. Farese RV,Sajan MP,Standaert ML.Insulin-sensitive protein kinases(atypical protein kinase C and protein kinase B/Akt):actions and defects in obesity and type Ⅱ diabetes.Exp Biol Med(Maywood),2005,230(9):593-605.
75. Tilg H,Diehl AM.Cytokines in alcoholic and nonalcoholic steatohepatitis.N Engl J Med,2000,343(20):1467-1476.
76. Vernochet C,Peres SB,Davis KE,et al.C/EBPalpha and the corepressors CtBP1 and CtBP2 regulate repression of select visceral white adipose genes during induction of the brown phenotype in white adipocytes by peroxisome proliferator-activated receptor gamma agonists. Mol Cell Biol, 2009,29(17):4714-4728.
77. Tomaru T, Steger DJ, Lefterova MI, et al. Adipocyte-specific expression of murine resistin is mediated by synergism between peroxisome proliferator-activated receptor gamma and CCAAT/enhancer-binding proteins. J Biol Chem,2009,284(10):6116-6125.
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