血管内皮细胞生长因子基因多态性与糖尿病肾病的相关性

目的研究血管内皮细胞生长因子（VEGF）基因多态性与糖尿病肾病（DN）的关系。方法单纯2型糖尿病（DM）组76例,DN组81例,健康对照（NC）组60例。UNIQ-10柱提取全血基因组DNA。标本基因型的判断用聚合酶链反应-限制性酶切片断长度多态性技术。Hardy-Weinberg平衡法检验各组基因频率的群体代表性。结果（1）DN组VEGF-460和＋405CC基因型频率和C等位基因频率明显高于DM组和NC组。（2）-460位点CC基因型DN患病率明显高于CT和TT基因型。＋405位点CC基因型DN患病率明显高于CG和GG基因型。（3）显示VEGF-460和＋405基因多态性均为DN发生的独立危险因素。结论（1）VEGF-460C/T基因多态性与DN发生有关。C等位基因可能是DN易感基因。（2）VEGF＋405G/C基因多态性与DN发生有关。C等位基因可能是DN的易感基因。     

Expression of vascular endothelial growth factor in patients with acute myocardial infarction

OBJECTIVE: The purpose of this study was to investigate the clinical significance of vascular endothelial growth factor (VEGF) in acute myocardial infarction (AMI). We also examined the involvement of peripheral blood mononuclear cells (PBMCs), which are a possible source of VEGF in AMI. BACKGROUND: VEGF is a potent endothelial cell-specific mitogen and could affect the outcome of AMI. METHODS: Thirty patients with AMI were used for this study. Serum and PBMCs were isolated from peripheral blood on days 1, 7, 14 and 21 after the onset of AMI. PBMCs were cultured at a density of 5 x 10(6) cells/ml for 24 h. VEGF levels in serum and the culture media were measured by enzyme-linked immunosorbent assay using a specific anti-human VEGF antibody. RESULTS: Serum VEGF levels elevated gradually after the onset of AMI and reached a peak on day 14. VEGF levels in the culture medium of PBMCs after incubation for 24 h (PBMC-VEGF) were maximally elevated 7 days after the onset. Maximum serum VEGF levels showed significant positive correlations with maximum creatine phosphokinase (CPK) levels (r = +0.70, p < 0.001), but maximum PBMC-VEGF levels did not correlate with maximum CPK levels. Patients showing improvement in left ventricular systolic function during the course of AMI showed significantly higher PBMC-VEGF levels than patients without improvement. CONCLUSIONS: The extent of myocardial damage contributes to the elevation of serum VEGF levels in AMI. VEGF produced by PBMCs may play an important role in the improvement of left ventricular function by promoting angiogenesis and reendothelialization after AMI.     

Increased blood vascular endothelial growth factor levels in patients with acute myocardial infarction

Vascular endothelial growth factor (VEGF) is a growth factor for vascular endothelial cells in vitro. The present study was designed to determine whether serum VEGF levels increase in patients with acute myocardial infarction (AMI) compared with patients with stable exertional angina and control subjects, and to examine the serial changes of serum VEGF levels in patients with AMI. We examined serum VEGF levels by using antibody prepared from serum immunized with human VEGF(121). The serum VEGF level (pg/ml) was higher (p < 0. 0001) on admission in the patients with AMI (177 +/- 19) than in those with stable exertional angina (61 +/- 7) and control subjects (62 +/- 6). The serum VEGF level (pg/ml) of the patients with AMI was 177 +/- 19 on admission, 125 +/- 9 on day 3, 137 +/- 11 on day 5, 242 +/- 18 at 1 week, and 258 +/- 22 at 2 weeks after admission. The value was higher on admission than on day 3 after admission (p = 0.014), the values were higher at 1 week and 2 weeks than on admission, on day 3, and 5 (p < 0.01). Furthermore, there were correlations between peak VEGF levels at 1 week or 2 weeks after admission and peak creatine kinase levels. The increase of VEGF on admission in the patients with AMI may be due to the hypoxia of acute myocardial ischemia. The elevation at 1 week and 2 weeks from the onset may cause the development of collateral circulation in relation to the healing of the infarction site.     

Quantitave and qualitative changes in vascular endothelial growth factor gene expression in glomeruli of patients with type 2 diabetes

OBJECTIVE: Vascular endothelial growth factor (VEGF) exists in three main splice variants, characterized by 121, 165 and 189 amino acids (VEGF 121, VEGF 165 and VEGF 189) and acts via two specific receptors: VEGF-R1 or Flt-1 and VEGF-R2 or KDR. VEGF plays an important role in the pathogenesis of diabetic retinopathy. This study examined the relationship between VEGF and its isoforms and the severity of diabetic nephropathy in type 2 diabetes. DESIGN: We evaluated the glomerular gene expression of VEGF and its receptors and studied the relationships with renal functional and structural parameters in type 2 diabetic patients. METHODS: Glomeruli from 17 kidney biopsies were microdissected; 14 out of 17 biopsies were also subjected to electron microscopic morphometric analysis to estimate glomerular structural parameters. VEGF mRNA was studied by comparative kinetic RT-PCR and real-time RT-PCR in order to identify the three different isoforms and to quantify VEGF, VEGF-R1 and VEGF-R2 mRNA levels. RESULTS: (i) Glomerular VEGF mRNA levels were inversely related to albumin excretion rate (r=-0.66, P=0.004); (ii) both the degree of mesangial and mesangial matrix expansion were inversely related to VEGF 165 mRNA levels (r=-0.73, P=0.005 and r=-0.64, P=0.017), and directly to VEGF 121 mRNA levels (r=0.74, P=0.003 and r=0.73, P=0.004); and (iii) VEGF and VEGF-R2 mRNA levels were directly related (r=0.62, P=0.033). CONCLUSIONS: These findings suggested that quantitative and qualitative changes in VEGF expression are present in type 2 diabetic patients with nephropathy and might be involved in the pathogenesis and progression of diabetic glomerulopathy.     

Expression of vascular endothelial growth factor in human myocardial infarction

Summary To define mechanisms that may influence collateral circulation and angiogenesis, we investigated vascular endothelial growth factor (VEGF) mRNA expression in human hearts. In non-ischemic human hearts, VEGF mRNA was not detected in vessels, but was found in cardiomyocytes. In hearts with myocardial infarction, the intensity of the VEGF signal was much higher in smooth muscle cells of arterioles adjacent to necrosis and in infiltrating macrophages than in myocytes around the site of the necrosis. This study suggests that levels of VEGF expression are high in smooth muscle cells and macrophages around infarcted areas after myocardial infarction and that VEGF may play a role in promoting collateral circulation and angiogenesis in human ischemic hearts.     

Association of RAGE gene polymorphism with vascular complications in Indian type 2 diabetes mellitus patients

Aims

The study was designed to evaluate the association of −374T/A and −429T/C polymorphism in the promoter region and Gly82Ser polymorphism in exon 3 region of RAGE gene with diabetic vascular complications in Indian population.

Methods

We screened 603 subjects which includes 176 healthy controls, 140 type 2 diabetes mellitus (T2DM) subjects without any vascular complications (DM), 152 T2DM subjects with microvascular complications (DM-micro) and 135 T2DM subjects with macrovascular complications (DM-macro) for −374T/A, −429T/C and Gly82Ser polymorphisms of RAGE gene. DNA isolated from the enrolled subjects were genotyped by PCR-RFLP. Logistic regression analysis was used to evaluate the association of single nucleotide polymorphisms (SNPs).

Results

The −429 T/C and Gly82Ser RAGE polymorphisms were found to be significantly associated with the development of macrovascular and microvascular complications, respectively, in T2DM subjects while −374A allele showed reduced risk towards the development of macrovascular complications. Further, −429T/C, −374T/A and Gly82Ser haplotype analysis revealed association of CTG haplotype with development of macrovascular complications while haplotype TAG was observed to be significantly protective towards development of macrovascular complications in T2DM subjects (OR = 0.617, p = 0.0202).

Conclusions

Our data indicates significant association of RAGE SNPs and haplotypes with vascular complications in North Indian T2DM subjects.     

IGF-I gene promoter polymorphism is a predictor of survival after myocardial infarction in patients with type 2 diabetes

OBJECTIVE: Previously we observed that non-carriers of the most common alleles of an IGF-I promoter polymorphism have low circulating IGF-I levels and an increased risk of developing myocardial infarction (MI), particularly in patients with type 2 diabetes. DESIGN: We investigated whether this IGF-I promoter polymorphism is associated with survival of type 2 diabetes in a Caucasian population aged 55 years and older. METHODS: The study was embedded in the Rotterdam Study, a prospective population-based cohort study. At baseline, 668 patients with type 2 diabetes were diagnosed, among which, 55 incident MI were ascertained during follow-up. For the present study, we used two genotype groups: non-variant carriers (homozygous for 192, 194, or 192/194 bp genotypes), and variant carriers. RESULTS: During a median follow-up of 8.8 years, 396 out of the 668 patients with type 2 diabetes (59.3%) died of various causes. The frequency of type 2 diabetes variant carrier and non-variant carriers was 28.7 and 71.3% respectively. The survival in patients with type 2 diabetes without an MI did not differ between the IGF-I genotype groups (hazard ratio (HR) = 0.8, 95% confidence interval (CI): 0.7-1.1, P = 0.1). In contrast, in those who developed an MI, variant carriers had a 2.4 times higher risk of mortality than non-variant carriers (95% CI: 1.2-4.8, P = 0.01). CONCLUSION: Our study suggests that genetically determined low IGF-I activity is an important determinant of survival in patients with type 2 diabetes who developed an MI. The IGF-I promoter polymorphism, therefore, may help to predict the future mortality risk in this group of patients.     

Elevated circulating levels of basic fibroblast growth factor and vascular endothelial growth factor in patients with acute myocardial infarction

Basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) have both shown strong angiogenetic effects in ischemic animal models and it has been reported that these growth factors were increased after acute myocardial ischemia. However, there have been few reports on the serum levels of bFGF and VEGF after acute myocardial infarction (AMI), in particular there has not been a comparative study of bFGF and VEGF in human subjects. The time course of circulating levels of bFGF and VEGF was examined in 36 patients with AMI who were within 24h of the onset of the AMI. The serum bFGF and VEGF levels of 50 age- and sex-matched healthy volunteers served as the baseline value. All the patients had undergone coronary angiography on the day of admission (Day 0), but prior to that the serum bFGF and VEGF levels were examined by enzyme-linked immunoassay. The serum bFGF and VEGF levels were also evaluated on Days 7, 14 and 28. Creatine kinase, myosin light chain I and troponin-T were measured subsequently and radionuclide examinations were performed during the early phase of AMI to determine the infarct size. The serum bFGF levels were significantly increased at Day 0 and were maintained until Days 7 and 14. Although serum VEGF levels at Day 0 were similar to the baseline values, they showed a remarkable increase by Days 7 and 14. A high serum level of bFGF was detected in the acute phase of AMI, and a later increase in VEGF was determined in the sub-acute phase, which suggest that these 2 growth factors play an important role at different time points of the reconstructing process of infarcted myocardial tissue.     

静脉应用血管内皮生长因子治疗急性心肌梗死的实验研究

目的探讨静脉应用血管内皮生长因子（ＶＥＧＦ）是否对急性心肌梗死大鼠有利。方法３３只成年雄性ＳＤ大鼠,结扎左冠状动脉后２４ｈ随机接受ＶＥＧＦ－肝素（治疗组）或肝素－生理盐水（对照组）治疗。ＶＥＧＦ（２μｇ／ｍｌ）加肝素５０Ｕ）或肝素－生理盐水（５０Ｕ／ｍｌ）每天经尾静脉注射,共用７ｄ（治疗组ｎ＝８;对照组ｎ＝１０）和１４ｄ（治疗组ｎ＝７;对照组ｎ＝８）。于结扎后９ｄ和１６ｄ分别测定血认动力学参数、心     

Circulating vascular endothelial growth factor in the normo- and/or microalbuminuric patients with type 2 diabetes mellitus

Relationship between serum vascular endothelial growth factor (VEGF) level and parameters of endothelial injury and/or dysfunction in patients with diabetes mellitus type 2 with or without microalbuminuria was investigated. Eighty-four diabetic patients were divided in two subgroups (42 each): normoalbuminuric (NAU) and microalbuminuric (MAU). Forty-two blood donors were in control group. Serum VEGF and plasma von Willebrand factor, soluble thrombomodulin, plasminogen activator inhibitor 1, thrombin-activatable fibrinolysis inhibitor (TAFI) and tissue plasminogen activator (t-PA) were measured using enzyme-linked immunosorbent assay in all subjects. VEGF was significantly higher in NAU compared to controls. The difference between MAU and controls was not statistically significant, but there was a trend toward significance. Only TAFI correlated with VEGF in MAU. An observed significant increase of serum VEGF level already in NAU suggests that serum VEGF could be a sensitive predictor of endothelial dysfunction in type 2 diabetes.     

血管内皮生长因子基因启动子区多态性与血管性痴呆的关系

目的研究血管内皮生长因子（VEGF）基因启动子区多态性与血管性痴呆（VD）发病的关系。方法选择中国北方汉族139例VD患者和150名健康对照者。用直接测序的方法对VEGF基因启动子区测序；用聚合酶链反摩限制性片段长度多态（PCR—RFLP）和直接电泳PCR产物的方法对VEGF基因启动子区多态性进行分型，并对所有标本进行载脂蛋白Ee4（ApoEe4）基因分型，将两组受试者分为携带和不携带ApoEe4基因亚组[ApoEe4（＋）和ApoEe4（-）组]。结果①中国北方汉族人群中的VEGF基因启动子区存在-2578C／A、-2549I／D和-1154G／A三个多态性位点。其中-2578C／A和-2549I／D存在显著的连锁不平衡，-2549I／D位点为18个碱基的插Ⅳ缺失。②多态性位点-2578C／A、-2549I／D的基因型频率、等位基因频率分布，VD组患者和健康对照组间差异无统计学意义（P〉0．05）。经过ApoEe4分层后，VD组和对照组基因型频率、等位基因频率分布差异也无统计学意义。-1154G／A位点，VD组患者的GG基因型频率、G等位基因频率分布明显高于健康对照组（P=0．037，P=0．018）。ApoEe4（-）的亚组中，这种差异同样存在（P=0．021，P=0．010）。用Logistic回归校正年龄、性别和ApoEe4基因后，显示-1154G／A位点GG基因型患者VD的发病风险是AA基因型的1．58倍（OR=1．58，95％CI：1．073—2．338，P=0．020）。（3）ApoEe4（-）的亚组中，-2549I／-1154A单体型在VD组中少于健康对照组（OR=0．536，95％CI．O．328～0．877，P=0．012）。-2549I／-1154G单体型在VD组中明显多于健康对照组（OR=1．785，95％CI：1．018～3．131，P=0．041）。在ApoEe4（＋）的亚组中差异无统计学意义。结论VEGF基因启动子区-1154G／A位点的GG基因型是VD发病的危险因素。此危险因素是独立于ApoEe4基因而影响VD发病的。     

急性心肌梗死患者外周血液单核细胞中血管内皮生长因子的表达

目的　探讨急性心肌梗死 ( AMI)后外周血单核细胞中血管内皮生长因子 ( VEGF)的表达及其与 AMI左心室收缩功能的关系。方法　分别抽取 2 5例行急诊经皮冠状动脉介入 ( PCI)治疗的 AMI患者发病后第 1、5、10和15 d的外周静脉血 ,分离、培养单核细胞 ,用酶联免疫吸附法 ( ELISA)测定 VEGF浓度。结果　单核细胞分泌的VEGF在 AMI后第 5 d即达高峰 ,显著高于对照组 ( 3 43 .2± 82 .5 pg/ml vs 14 3 .3± 2 4.2 pg/m l,P<0 .0 5 )。单核细胞分泌的 VEGF峰植与肌酸磷酸激酶 ( CK)峰值无显著相关。 PCI术后左心室收缩功能改善的 AMI患者 ,其单核细胞分泌的 VEGF水平显著高于左心室收缩功能未改善者 ( 867.6± 113 .1pg/ml vs2 3 4.8± 82 .4pg/ml,P<0 .0 5 )。结论　AMI患者 PCI术后单核细胞产生的 VEGF与左心室收缩功能改善有关     

Association between plasma visfatin and vascular endothelial function in patients with type 2 diabetes mellitus

Visfatin is a newly identified adipocytokine that mimics insulin action. However, the pathophysiological role of visfatin in diabetic patients is not fully understood. The main purpose of this study was to investigate the association of plasma visfatin with endothelial function in patients with type 2 diabetes mellitus. In addition, the relationships of visfatin with oxidative stress, low-grade inflammation, atherosclerosis, adiponectin, plasma renin activity, and aldosterone were also explored, and the effect of pioglitazone on visfatin was examined. Visfatin levels were measured in 80 patients with type 2 diabetes mellitus and in 28 age-matched healthy subjects. Endothelial function was evaluated by using flow-mediated vasodilatation (FMD), oxidative stress was assessed by the level of urinary 8-iso-prostaglandin F2alpha, and atherosclerosis and inflammation were measured by using the intimal-medial complex thickness and the levels of high-sensitivity C-reactive protein and fibrinogen. Pioglitazone was administered for 12 weeks at a dose of 30 mg/d in a further 20 patients with type 2 diabetes mellitus. There was a significant negative correlation between the log10-transformed (log) plasma visfatin concentration and FMD or creatinine clearance (R=-0.2672, P=.0167; R=-0.2750, P=.0136). Log visfatin was also positively correlated with log urinary albumin excretion (R=0.2305, P=.0397). In addition, it was also found that visfatin had a significant negative correlation with plasma aldosterone (R=-0.2432, P=.0297). In stepwise regression analysis, creatinine clearance, log aldosterone, FMD, and sex showed a significant association with log visfatin (P=.0040, P=.0069, P=.0444, and P=.0487, respectively), and log 8-iso-prostaglandin F2alpha showed a tendency for an association (P=.0515). Pioglitazone therapy did not affect the visfatin concentration in the 20 pioglitazone-treated patients with diabetes, although a significant elevation of visfatin was obtained in a subgroup of 11 female patients (P=.0381). In conclusion, the current study showed that visfatin is negatively associated with vascular endothelial function evaluated by FMD and creatinine clearance, and positively associated with log urinary albumin excretion. Visfatin was also negatively correlated with circulating aldosterone. Pioglitazone therapy for 12 weeks did not affect the plasma visfatin concentration significantly in all diabetic patients, but a significant elevation in visfatin was obtained in women only.     

Association of endothelial lipase Thr111Ile polymorphism with proliferative retinopathy in type 2 diabetes patients

AimOur previous study demonstrated that the endothelial lipase (EL) C.584C>T polymorphism (rs2000813, p.Thr111Ile) was significantly associated with diabetic retinopathy (DR). The present work was conducted to see if this specific variant of the EL gene was more specifically linked to the severity of DR.MethodsThis retrospective cohort study was based on a review of the institutional charts of 287 type 2 diabetes patients (mean age = 59.7 years; mean BMI = 29.0 kg/m²; mean HbA_1c = 8.4%) genotyped for the EL C.584C>T polymorphism (rs2000813, p.Thr111Ile). The stage of DR was also determined for each genotype (CC, CT, TT).ResultsOn univariate analysis, the minor allele homozygote TT variant was significantly associated with severe DR (OR: 4.3; 95% CI: 1.4, 13.1) compared with the major CC homozygote. No significant result was found for the CT heterozygote. Multivariate analysis revealed an increased risk for TT homozygotes to present with severe non-proliferative DR (OR: 8.09; 95% CI: 1.23, 53.1) or proliferative DR. Other associations were not significant.ConclusionMinor allele homozygosity for this EL variant (c.584C>T) could be a significant risk factor for developing severe, sight-threatening disease due to proliferative DR. Further prospective studies of this EL polymorphism in a larger population sample are needed to confirm these results.     

血管内皮生长因子基因多态性与糖尿病视网膜病变的相关性

目的探讨血管内皮生长因子（VEGF）-460C/T基因多态性与糖尿病视网膜病变（DR）的相关性。方法病史超过10年的2型糖尿病（T2DM）患者204例,分为非增殖型视网膜病变组（NP-DR,65例）、增殖型视网膜病变组（PDR,64例）及单纯2型糖尿病组（DM,75例）。用PCR-RFLP方法检测各组基因型,比较各组基因型和等位基因的频率。结果DR组VEGF-460位点TT基因型频率显著低于DM组（P〈0.01）,C等位基因频率显著高于DM组（P〈0.01）;NPDR组与PDR组基因型和等位基因频率差异无统计学意义（P〉0.05）。DM中CC、CT、TT基因型的DR发生率分别为69.8%、68.9%和42.2%,CC和CT基因型的DR发生率显著高于TT基因型（P〈0.01）。结论VEGF-460C/T多态性与DR的发生发展有关,C等位基因可能是DR的易感基因。     

乙酰辅酶A羧化酶β基因多态性与2型糖尿病易感性关系的初步研究

目的 研究上海地区汉族人群中乙酰辅酶A羧化酶B（ACC-β）的2个单核苷酸多态性（SNP）位点与2型糖尿病（T2DM）易感性的关系。方法 采用等位基因专一性实时PCR的方法对上海地区438例T2DM患者及328名正常对照者（NC）ACC-β基因的2个SNP位点进行分析。结果 （1）ACC-β基因16号内含子区存在SNP位点116／73C＞T，T2DM组和NC组两组间基因型频率比较，差异有统计学意义（P=0．031）。且CC型＋CT型与TT型的分组比较在两组间的差异也有统计学意义（P=0．027）。（2）ACC-B基因16号内含子区还存在另-SNP位点：116／288A〉G，T2DM组G等位基因的频率高于NC组，但无统计学意义。结论 在上海地区的汉族人群中，ACC-β基因可能是T2DM的易感基因之一，其16号内含子区的116／73C＞T多态性（rs2268393）可能与T2DM的发病相关。     

磷酸烯醇式丙酮酸羧激酶基因多态性与2型糖尿病的相关性研究

目的　研究华东地区汉族人群中磷酸烯醇式丙酮酸羧激酶 (PCK1)基因的单核苷酸多态性(SNP)与 2型糖尿病的相关性。方法　采用直接测序法对PCK1基因作SNP筛查 ,并按病例 对照研究选用直接测序法对改变氨基酸编码的cSNP作进一步的基因分型。结果　共检出 11个SNPs ,其中出现在启动子区 3个 (P/-10 99G→A ,P/-969G→A ,P/-2 3 4G→C) ,内含子区 2个 (I3 /3 79A→T ,I5/85C→G) ,外显子区 6个SNP(E2 /69C→T ,E4/47C→T ,E4/56C→T ,E4/83C→T ,E4/14 4G→C ,E7/179G→A) ,而改变氨基酸编码的cSNP只有E4/14 4G→C ,即Leu184Val。进一步基因分型后显示 ,2型糖尿病组中GG :10 6,GC :77,CC :8,而正常对照组中GG :13 0 ,GC :58,CC :3 ,这两组间的基因型频率差异有显著性 (P <0 .0 5) ,而且两组间等位基因频率差异也有显著性 (P <0 .0 1)。结论　PCK1基因的Leu184Val多态性与华东地区汉族人群 2型糖尿病的发生可能存在一定相关性     

对氧磷酶2基因多态性与2型糖尿病肾病的相关性研究

采用PCRRFLP法检测了327例不同程度白蛋白尿的2型糖尿病患者对氧磷酶2(PON2)基因第9外显子的C311S多态性。结果显示,PON2基因C311S多态性与糖尿病早期肾病的发生显著相关,C等位基因可能是2型糖尿病早期肾病一个独立的危险因素。