血管内皮生长因子基因多态性与糖尿病肾病的相关性研究

目的 探讨血管内皮生长因子(VEGF)-634G／C基因多态性与糖尿病肾病(DN)的关系。方法 运用PCR-限制性多态性片段长度(RFLP)技术检测98例健康对照者和216例2型糖尿病患者[其中DN患者104例，单纯2型糖尿病(DM)患者112例]的基因型，比较各组的基因型和等位基因频率。结果 (1)CC基因型者血清VEGFT水平高于CG及GG型者；(2)DN组CC基因型和C等位基因频率显著高于DM组和正常对照组；(3)与GG型和CG型组相比，CC型组DN的发生率明显上升；(4)Logistic回归分析显示VEGF、VEGF基因多态性、收缩压(SBP)、HbA1c、LDL-C、体重指数(BMI)是DN的危险因素。结论 VEGF-63gG／C多态性与2型DM伴发肾病的发生有关，C等位基因可能是DN的易感基因。     

维生素D受体基因BsmI多态性与2型糖尿病肾病的关系

目的 研究维生素D受体(VDR)基因BsmI位点多态性与汉族人群2型糖尿病肾病(DN)的关系.方法 应用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)技术检测304例2型糖尿病患者(DM组)及100例健康体检者(NC组)VDR Bsml位点基因型和等位基因频率.根据尿白蛋白情况将DM组分为非糖尿病肾病组(DN0组,122例)、微量白蛋白尿组(DN1组,87例)、大量白蛋白尿组(DN2组,95例).83例病程5年以上仍未出现肾病的DM患者纳入L-NDN组;64例起病1年内即出现肾病的DM患者纳入EDN组.结果 DM组BB+Bb基因型和B等位基因频率均高于NC组(x2=7.088,P=0.008;x2=5.865,P=0.015).DN2组BB+Bb基因型和B等位基因频率高于NC组(x2=14.287,P=0.000; x2=12.621,P=0.000)及DN0组(x2=8.063,P=0.005;x2=8.173,P=0.004).其余组间差异均无统计学意义.EDN组BB+Bb基因型和B等位基因频率均显著高于L-NDN组(x2=7.228,P=0.007;x2=5.853,P=0.016).B等位基因阳性DN患者的尿白蛋白排泄率显著高于B等位基因阴性DN患者,差异有统计学意义(P＜0.01).BsmI位点基因型与DN发生密切相关.B等位基因阳性是DN发生及早发的危险因素(OR=2.004;0R=2.394).结论 VDRBsmI基因多态性与DN易感性相关.B等位基因阳性患者更易出现大量白蛋白尿及早期发生肾病.     

血管紧张素Ⅰ转化酶及血管紧张素Ⅱ受体1型基因多态性与糖尿病肾病的关联研究

目的探讨血管紧张素I转化酶(angiotensin converting enzyme,ACE)基因I/D多态性及血管紧张素Ⅱ受体1型(angiotensinⅡtype 1 receptor,AT1R)基因A1166C多态性与糖尿病肾病的关系。方法选择2013年4月至2015年10月在广东医科大学附属医院、东莞市塘厦医院和东莞市大朗医院住院2型糖尿病患者,研究对象分为2型糖尿病组(T2DM组)97例和正常对照组50例,其中T2DM组根据有无肾病分为52例糖尿病肾病(DN)组和45例糖尿病非肾病(非DN)组;采用标准的酚-氯仿-蛋白酶K法提取外周血DNA;分别用聚合酶链反应(PCR)技术、PCR-限制性片段长度多态性技术(PCR-RFLP)对ACE I/D、ATlR Al166C多态性进行检测。遗传平衡吻合度检验用HardyWeinberg平衡法。采用χ~2检验分析基因多态性与DN的相关性。结果 DN组和非DN组基线资料无统计学差异(P0.05)。经χ~2检验,DN组DD基因型频率32.6%(17/52)明显高于非DN组DD基因型频率20.0%(9/45)(χ~2=6.62,P0.05);DN组D等位基因频率58.7%(61/104)明显高于非DN组D基因型频率41.1%(37/90)(χ~2=5.94,P0.05)。DN组与非DN组AT1R Al166C各基因型分布频率无显著性差异(χ~2=0.22,P0.05);各等位基因频率分布亦无明显差异(χ~2=0.21,P0.05)。对照组与T2DM组ACE I/D和ATlR Al166C基因多态的基因型分布和等位基因频率均无显著性差异(P0.05)。结论 ACE基因I/D多态性与DN有关,携带DD基因型和D等位基因的T2DM患者是DN的易感人群。AT1R基因A1166C多态性与T2DM并发DN无关。     

大肠癌发病风险与细胞间黏附分子-1基因多态性的关系

目的 探讨中国华北地区汉族人群中细胞间黏附分子-1(ICAM-1)基因编码区单核苷酸多态性(SNPs)与大肠癌发病风险的关系.方法 采用序列特异-聚合酶链反应(PCR-SSP)方法检测87例大肠癌患者(大肠癌组)和102例正常对照者(对照组)ICAM-1基因型和等位基因频率的分布.结果 大肠癌组和对照组ICAM-1-241G/R多态的基因型均为G/G,未检测出R241等位基因.ICAM-1-469K/E多态3种基因型频率(K/K,K/E,E/E)在大肠癌组和对照组中分别是57.47%(50/87)、32.18%(28/87)、10.35%(9/87)和42.16%(43/102)、43.14%(44/102)、14.70%(15/102),与K/E+E/E基因型比较,K/K基因型罹患大肠癌的风险明显增加(OR=1.85,x2=4.406,95%CI:1.04～3.31,P＜0.05);与E等位基因比较,K等位基因携带者增加大肠癌的发病风险(OR=1.58,x2=4.194,95%CI:1.02～2.46,P＜0.05).ICAM-1-469K/E多态的K/K基因型在大肠癌组中与肿瘤分化程度有关(x2=4.564,P＜0.05);而与临床其他病理参数包括性别、年龄、肿瘤部位及Dukes分期无关(P＞0.05).结论 ICAM-1-241G/R多态在我国华北地区汉族人群中不存在多态性;ICAM-1-469K/E多态K等位基因或K/K基因型的存在可明显增加大肠癌的发病风险;ICAM-1-469K/E多态K/K基因型与肿瘤分化程度有关.     

肾素-血管紧张素系统基因多态性与2型糖尿病患者慢性肾脏病的相关性研究

目的探讨血管紧张素原(AGT)及血管紧张素Ⅱ1型受体(AT1R)基因多态性与2型糖尿病患者慢性肾脏病(CKD)的关系。方法将76例患CKD的2型糖尿病患者,根据肾穿刺活检病理分为糖尿病肾病(DN)组(28例)、非糖尿病性肾病(NDRD)组(30例)和DN合并NDRD组(18例);另外选择30名健康体检者作为正常对照组。采用聚合酶链反应-限制性片段长度多态性技术方法检测上述研究对象的AGT基因M235 T多态性和AT1R基因A1166C多态性。结果NDRD组的主要病理类型是为膜性肾病和IgA肾病,DN+NDRD组NDRD的主要病理类型为膜性肾病、IgA肾病和高血压性肾小动脉硬化,两组NDRD的病理类型均无显著性差异(P0.05)。DN组和DN+NDRD组AGT基因M235 T-TT基因型频率明显高于NDRD组和正常对照组(P0.05),T等位基因频率明显高于NDRD组(P0.05)和正常对照组(P0.01),NDRD组和正常对照组比较以及DN组和DN+NDRD组比较AGT-TT基因型和T等位基因频率均无明显差异(P0.05)。各组AT1R基因A1166C多态性无显著性差异。AGT基因M235 T-TT基因型为2型糖尿病肾脏疾病患者eGFR下降的危险因素。结论AGT基因M235 T-TT基因型以及T等位基因与2型糖尿病患者DN及DN合并NDRD的发生有关,与NDRD的发生无关。AGT基因M235 T-TT基因型是2型糖尿病CKD患者肾功能减退的易感因素。AT1R基因多态性与2型糖尿病患者肾脏疾病的发生发展无关。     

醛糖还原酶基因5＇端(A-C)n的多态性与Ⅱ型糖尿病患者糖尿病肾病易感性的关系

目的探讨Ⅱ型糖尿病（DM）醛糖还原酶（AR）基因5’端的（AC）n的多态性与糖尿病肾病（DN）易感性的关系。方法应用32PdCTP掺入聚合酶链反应（PCR）,PCR产物6％甲酰胺聚丙烯酰胺凝胶电泳及放射性自显影的方法,分析了113例Ⅱ型DM组和42例正常对照（CON）组醛糖还原酶（AR）基因5’端的（AC）n的多态性与糖尿病肾病易感性的关系。结果DM组和对照组均发现7种等位基因（Z－6～Z＋6）,Z－2等位基因频率在DN组高于无微血管并发症组（NDC）和CON组;Z＋2等位基因频率在DN组低于NDC和CON组,P＜0．05。结论中国北方汉族Ⅱ型DM患者AR基因5’端（AC）n存在7种等位基因（Z－6～Z＋6）。Z－2等位基因可能是DN的易感基因,Z＋2等位基因可能为其保护基因。     

Klotho基因多态性与糖尿病肾病中医证候的相关性分析

目的:研究Klotho基因多态性与2型糖尿病、糖尿病肾病发病风险及糖尿病肾病中医证候之间的关联,筛选可能的易感基因型,以期对潜在高危患者及时防治。方法:采用PCR联合直接测序技术检测56例T2DM,67例T2DN,55例健康对照组的Klotho G-395A位点单核苷酸多态性。结果:G-395A不同基因型及等位基因分布频率在病例组和健康对照组间差异无统计学意义(P=0.274;P=0.298);而在T2DM组和T2DN组间比较,差异有统计学意义(P=0.025;P=0.015);Logistic回归分析结果:携带A等位基因、高血压病史、糖尿病病程及糖化血红蛋白与DN的发生有统计学关联(P0.01)。T2DN组三种中医证候间GA+AA基因型及A等位基因频率分布有显著差别,具有统计学意义(P=0.013;P=0.037)。结论:本课题未发现G-395A位点多态性与T2DM的发病风险存在关联性;携带A等位基因可能是湖北地区汉族人群T2DM患者合并肾损害的独立危险因素之一,而GA及AA基因型可能是糖尿病肾病的遗传易感危险基因型。携带A等位基因可能是脾肾阳虚型糖尿病肾病的风险因素之一。     

内皮细胞固有型一氧化氮合酶基因内含子4插入/缺失多态性与糖尿病肾病的关系

目的：对天津地区汉族人内皮细胞固有型一氧化氮合酶（ecNOS)基因内含子4的插入／缺失多态性（ecNOS4b/a)与2型糖尿病肾病（DN）的关联性进行研究。方法：应用PCR－小卫星DNA多态性分析技术对ecNOS4b/a基因型分布进行检测。包括正常对照组70例，2型糖尿病无DN组48例，2型糖尿病有DN无慢性肾功能不全（CRF）组35例，2型糖尿病DN有CRF组45例和非DN导致的CRF组58例。结果：（1）发现1例罕见基因型（女性DN无CRF患者），为447bp 420bp杂合子。（2）2型糖尿病无DN组a等位基因频率高于正常对照组，差异无显著性意义（χ^2=1.672,P=0.196).(3)2型糖尿病伴DN组a等位基因频率低于2型糖尿病无DN组，差异无显著性意义（χ^2=1.082,P=0.298)。（4）DN无CRF组与DN伴CRF组等位基因频率相近（校正χ^2=0.002,P=0.967)。（5）DN伴CRF组a等位基因频率低于其它原因导致的CRF组，差异有显著性意义（χ^2=0.002,P=0.967)。（5）DN伴CRF组a等位基因频率低于其它原因导致的CRF组，差异有显著性意义（χ^2=4.360,P=06037)。结论：a等位基因可能不是天津汉族人DN的危险因素；天津地区汉族健康人群a等位基因频率低于日本；ecNOS在DN导致的CRF中的作用可能与非DN CRF不同。     

2型糖尿病肾病患者p22phox基因多态性的研究

目的研究NADPH氧化酶p22phox亚基基因多态性与中国上海汉族人群2型糖尿病肾病（DN）相关性。方法应用限制性片断长度多态性（RFLP）-PCR方法对105例健康对照组和194例2型糖尿病（DM）患者（其中71例DN）进行p22phox亚基C242T、A640G基因型检测。同时检测其身高、体重、血压、血脂、空腹血糖及胰岛素、HbAlc的水平。结果DN组CT＋TT基因型频率明显高于2型DM和对照组（26．76％比17．07％、3．81％，P=0．0002）；DN组T等位基因频率明显高于2型DM和对照组（22．54％比13．42％、2．86％，P=0．0001）；3组间AA基因型频率与A等位基因频率差异无统计学意义。多元回归分析显示，T242等位基因、收缩压、空腹血糖、HbAlc、β细胞功能指数（Homa-IS）是DN的危险因素。结论p22phox亚基T242等位基因变异可能是中国上海地区汉族人群DN的易感基因：而p22phox亚基A640G基因多态性与上述人群DN无相关性。T242等位基因、收缩压、空腹血糖、HbAlc、Homa-IS是DN的危险因素。     

MTHFR基因多态性与2型糖尿病肾病相关性的研究

目的 研究亚甲基四氢叶酸还原酶（MTHFR）基因多态性与2型糖尿病肾病（DN）易感性的关系.方法 选择111例山西地区汉族人2型糖尿病患者,其中糖尿病肾病（DN＋）组56例,糖尿病非肾病（DN-）组55例,运用聚合酶链式反应-限制性片段长度多态性（PCR-RFLP）技术并结合琼脂糖凝胶电泳的方法检测111例患者的MTHFR基因多态性,测定各组间基因型频率和等位基因频率.结果 纯合基因型TT、T等位基因在DN＋组（21.43％,46.43％）的频率均明显高于DN-组（7.27％,29.09％）,差异均具有统计学意义（P＜0.05）.无论是在DN＋组还是DN-组中,TT基因型患者血同型半胱氨酸（Hcy）平均水平均大于CC基因型和CT基因型患者,DN＋组血浆Hcy水平明显高于DN-组,差异均具有统计学意义（P＜0.05）.在叶酸浓度≤6.92nmol/L时,DN＋组（24.24％,48.49％） TT型发生率及T等位基因频率明显高于DN-组（3.70％,25.93％）（P＜0.05）,当叶酸浓度＞6,92nmol/L时,DN＋组TT型发生率及T等位基因频率与DN-组无差异（P＞0.05）.结论 MTHFR基因C677T多态性与糖尿病肾病（DN）发生具有相关性,突变的T等位基因是DN易感基因,但其影响效果受叶酸浓度的影响.     

Higher frequency of apolipoprotein E2 allele in type 2 diabetic patients with nephropathy in Taiwan

BACKGROUND: Diabetes is one of the major causes of end stage renal failure in Taiwan. Previous studies have indicated that lipid abnormalities might contribute to the development and progression of diabetic nephropathy (DN). Apolipoprotein E (apo E) regulates the metabolism of lipoproteins. Three different apo E alleles (epsilon2, epsilon3 and epsilon4) produce apo E isoproteins, apo E2, apo E3 and apo E4. Thus, the apo E gene polymorphism may be associated with DN. METHODS: To investigate the distribution of apo E genotype in type 2 diabetic Taiwanese, we examined 314 patients with type 2 diabetes (100 without nephropathy and 214 with nephropathy) and 150 age-matched normal controls. The diagnosis of nephropathy was made when daily protein loss exceeded 500 mg. The apo E gene polymorphism was analyzed by polymerase chain reaction. RESULTS: Our study found that the frequency of apo E2 allele was significantly higher in the diabetics with nephropathy- non-dialysis group (7.7%) and diabetics with nephropathy- dialysis group (7.7%) than in normal controls (1.7%) (p < 0.001) and diabetics without nephropathy (2.0%) (p < 0.01). The E3 and E4 allele frequencies were not significantly different among groups. CONCLUSION: These findings imply that apo E polymorphism is apparently related to the development of DN in type 2 diabetes in Taiwan.     

Increased frequency of angiotensin-converting enzyme DD genotype in patients with type 2 diabetes in Taiwan.

BACKGROUND: Diabetes is one of the major causes of end-stage renal failure in the Taiwanese population. Previous studies have shown that angiotensin-converting enzyme (ACE) inhibitor can improve glucose utilization and suppress hepatic glucose production and the renin-angiotensin system may play an important role in the initiation and progression of diabetic nephropathy. Thus, ACE gene polymorphism may be associated with type 2 diabetes and diabetic nephropathy. METHODS: To investigate the distribution of ACE-I/D genotype in type 2 diabetes and diabetic nephropathy, we examined 336 patients with type 2 diabetes (157 without nephropathy and 179 with nephropathy) and 263 age-matched normal controls. The diagnosis of nephropathy was made when daily protein loss exceeded 500 mg. ACE gene polymorphism was analysed by use of polymerase chain reaction. RESULTS: Our study revealed that the frequency of the D allele of the ACE gene was 29.3% in normal controls. The frequency of ACE DD genotype was significantly higher in type 2 diabetics compared with normal controls (18.2 vs 9.1%, P<0.01). The frequency of ACE DD genotype in patients with diabetic nephropathy was significantly higher than in patients without nephropathy (22.3 vs 13.4%, P<0.05). To determine whether ACE gene polymorphism was associated with the severity of diabetic nephropathy, we divided patients with diabetic nephropathy into dialysis and non-dialysis groups. The frequency of ACE DD genotype in the dialysis group was significantly higher than in non-dialysis group (28.7 vs 15.3%, P<0.05). CONCLUSION: Our results indicate that the frequency of ACE DD genotype is markedly higher in patients with type 2 diabetes, and the ACE DD genotype is significantly associated with diabetic nephropathy.     

Angiotensinogen gene M235T polymorphism is not associated with diabetic nephropathy

BACKGROUND.: There is agreement that a family history of hypertension (HT),is a predictor for the risk of diabetic nephropathy (DN) inpatients with type 2 diabetes, and possibly also type 1 diabetes.It follows that genes related to the risk of hypertension mustalso be considered candidate genes for DN. The 235T allele ofthe angiotensinogen gene was found to be related to primaryHT. METHODS.: To examine whether it is predictive for DN as well, we examinedthe angiotensinogen gene polymorphism in 230 healthy local controls,423 patients with type 1 diabetes (n=180 with DN; n=243 withoutDN) and 663 patients with type 2 diabetes (n=310 with DN; n=353without DN). The angiotensinogen gene M235T polymorphism wasdetermined using PCR amplification. RESULTS.: The following results were obtained (i) no significant differenceof genotype distribution (type 1: MM/MT/TT(%) 27.6/57.2/15.2vs. 27.2/56.1/16.7 (P=0.92); type 2: MM/MT/TT (%) 31.7/48.2/20.1vs. 32.9/46.8/20.3 (P=0.93)) or allele frequencies (type 1:M 0.56 vs. 0.55 (P=0.795); type 2: M 0.56 vs. 0.56 (P=0.86))was found, between diabetic patients with or without DN, (ii)no difference was found between normotensive and hypertensivediabetic patients. CONCLUSION.: The data argue against a role of the angiotensinogen gene M235Tpolymorphism in the manifestation of diabetic nephropathy orhypertension in diabetic patients.     

对氧磷酯酶基因多态性与糖尿病肾病的关系

目的探讨对氧磷酯酶2(PON2)基因A148G多态性与糖尿病肾病的关系.方法(1)用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)分析法探查PON2基因A148G多态性在正常对照组、单纯2型糖尿病组、糖尿病肾病组中的基因频率分布;(2)放射免疫法检测血清免疫反应性胰岛素(IRI)、C肽(C-P)水平.结果(1)糖尿病肾病组GG基因型和G等位基因频率明显高于单纯2型糖尿病组(X2=4.26 P＜0.05,X2=4.89P＜0.05)和正常对照组(X2=4.79 P＜0.05,X2=5.49P＜0.01);(2)基因型为GG的糖尿病患者空腹血糖浓度高于基因型为GA和AA的糖尿病患者的空腹血糖浓度(F=3.90 P＜0.05,F=4.23 P＜0.05);(3)Logistic回归分析表明GG基因型是糖尿病肾病的独立变异危险因素(P＜0.05).结论PON2基因多态性与糖尿病肾病的发生有关.     

A polymorphism in the gene for the atrial natriuretic peptide and diabetic nephropathy

Background: Atrial natriuretic peptide is involved in blood pressure regulation via its vasodilating and natriuretic actions. Since diabetic nephropathy and hypertension are closely related, ANP is a reasonable candidate gene for diabetic nephropathy (DN). Methods: We genotyped 410 patients with type I diabetes (without DN n=307; with DN n=103) and 658 patients with type II diabetes (without DN n=464; with DN n=194). In the patients the duration of diabetes was at least 10 years. Diabetic nephropathy was defined as urinary albumin excretion of ⩾30 mg/24 h. The Hpa II polymorphism in intron 2 of the ANP gene was determined using PCR amplification followed by restriction digest. Alleles were separated on agarose gels stained with ethidium bromide. Results: We compared genotype distribution and allele frequencies between patients with and without nephropathy. No significant difference was observed either in type I (allele frequency without DN H1, 0.02/H2, 0.98 vs with DN H1, 0.05/H2, 0.95; P=0.132) or in type II diabetes (allele frequency without DN H1, 0.04/H2, 0.96 vs with DN H1, 0.05/H2, 0.95; P=0.551). Conclusions: The polymorphism in the gene for the atrial natriuretic peptide does not seem to play a major role in the development of diabetic nephropathy in either type I or in type II diabetes.     

Relationship of p22phox C242T polymorphism with nephropathy in type 2 diabetic patients

BACKGROUND: In this case-control study, we investigated the possible involvement of the p22phox C242T polymorphism in the development and progression of diabetic nephropathy (DN) in 535 Caucasian Brazilians with type 2 diabetes. We also evaluated the effects of the interaction of the C242T polymorphism with smoking and hypercholesterolemia on the susceptibility to nephropathy. METHODS: Genotype analysis was performed using polymerase chain reaction (PCR) followed by digestion with restriction enzyme. Logistic regression analysis was used to control for independent risk factors associated with nephropathy. RESULTS: The genotype frequencies in patients with overt DN (CC/CT/TT: 0.36/0.47/0.17) were not significantly different from those of diabetic individuals with normoalbuminuria (0.47/0.41/0.12) or microalbuminuria (0.42/0.48/0.10) (p=0.214). Likewise, there were no differences in the T allele frequency among patients with normoalbuminuria, microalbuminuria or overt DN (0.33, 0.34 and 0.40, respectively; p=0.111). However, the T allele was found to be more frequent among smokers with overt nephropathy (macroalbuminuria and/or in dialysis) than those who had normoalbuminuria (43 vs. 32%, p=0.045). The multiple logistic regression analysis confirmed that the CT+TT genotypes were independently associated with a higher risk of having overt nephropathy among smokers [odds ratio (OR)=6.76, 95% confidence interval (95% CI) 1.83-25.02]. CONCLUSIONS: Our study shows a gene-environment interaction associated with the increased risk of DN progression in Caucasian Brazilian smokers with type 2 diabetes. Further studies should be performed to clarify whether it exists, and to what extent there is a relationship between the p22phox C242T polymorphism and DN.