【摘要】：目的： 研究湖南地區(qū)2型糖尿病視網(wǎng)膜病變(diatetic retinopathy,DR)遺傳及環(huán)境易感因素；篩選糖尿病視網(wǎng)膜病變發(fā)生的高危人群。 (1)采用薈萃分析方法,搜尋黃種人中與糖尿病視網(wǎng)膜病變發(fā)生相關(guān)的血管內(nèi)皮生長(zhǎng)因子(vascular endothelial growthfactor,VEGF)單核苷酸多態(tài)性位點(diǎn)。 (2)探討湖南地區(qū)人群中VEGF單核苷酸多態(tài)性位點(diǎn)與糖尿病視網(wǎng)膜病變易感性的關(guān)聯(lián)性。 (3)探討血清中VEGF蛋白總量,亞型VEGF165及其異構(gòu)體VEGF165b含量與糖尿病視網(wǎng)膜病變的相關(guān)性。 (4)采用病例對(duì)照及l(fā)ogistic回歸分析比較VEGF單核苷酸多態(tài)性位點(diǎn)、VEGF蛋白及其他人口學(xué)特征和生化指標(biāo),探討糖尿病視網(wǎng)膜病變發(fā)生的高危因素。 方法： (1)回顧2012年3月前有關(guān)VEGF單核苷酸多態(tài)性與糖尿病視網(wǎng)膜病變的所有英文文獻(xiàn)。系統(tǒng)性檢索及獲取原始文獻(xiàn)、制定納入及排除標(biāo)準(zhǔn)；將單核苷酸多態(tài)性位點(diǎn)與糖尿病視網(wǎng)膜病變相關(guān)的實(shí)驗(yàn)數(shù)據(jù)進(jìn)行薈萃分析。 (2)根據(jù)薈萃分析顯示黃種人中有意義的VEGF單核苷酸多態(tài)性位點(diǎn),采用病例對(duì)照方法,使用限制性片段長(zhǎng)度多態(tài)性聚合酶鏈反應(yīng)(PCR-RFLR)檢測(cè)其在湖南地區(qū)正常人、糖尿病無(wú)視網(wǎng)膜病變患者(diabetic without retinopathy, DWR)、鐕糖尿病視網(wǎng)膜病變患者(diabetic retinopathy, DR)的分布,通過(guò)卡方檢驗(yàn)統(tǒng)計(jì)分析其在各組間的分布差異。 (3)采用病例對(duì)照方式,使用酶聯(lián)免疫吸附測(cè)定法檢測(cè)湖南地區(qū)正常人,糖尿病無(wú)視網(wǎng)膜病變患者,糖尿病視網(wǎng)膜病變患者的血清中VEGF總量,亞型/EGF165及其異構(gòu)體VEGF165b含量,通過(guò)成組T檢驗(yàn)及線性回歸分析統(tǒng)計(jì)分析其含量在各組間的分布差異,及與、/EGF單核苷酸多態(tài)性位點(diǎn)及環(huán)境因素間的關(guān)聯(lián)性。 (4)采用病例對(duì)照方式,統(tǒng)計(jì)糖尿病無(wú)視網(wǎng)膜病變患者,糖尿病視網(wǎng)膜病變患者的人口學(xué)特征及生化指標(biāo)的相關(guān)數(shù)據(jù)及VEGF單核苷酸多態(tài)性位點(diǎn)及VEGF蛋白表達(dá),進(jìn)行成組T檢驗(yàn)及l(fā)ogistic回歸分析,探討糖尿病視網(wǎng)膜病發(fā)生的高危易感因素。 結(jié)果： (1)薈萃分析結(jié)果： VEGF rs699947共顯性模型-雜合型／野生型(CAVs CC)與DR發(fā)病風(fēng)險(xiǎn)存在相關(guān)性(I2=64%, OR=1.2795%CI [1.05,1.54], P=0.02),尤其在黃色人種中(I2=38%, OR=1.4695%CI [1.17,1.82], P=0.0007)。其它模型與DR發(fā)病無(wú)顯著相關(guān)性(P0.05) VEGF rs1570360各模型在各色人種中與DR的發(fā)病無(wú)顯著相關(guān)性(P0.05)。 VEGF rs2010963亞組分析：黃色人種組隱性模型(CC V s GC+GG)顯示與DR的發(fā)病相關(guān)(I2=60%, OR=1.2895%CI [1.01,1.63], P=0.04); PCR-RFLR測(cè)序方式組隱性模型(CC V s GC+GG)與DR的發(fā)生明顯相關(guān)(I2=46%, OR=1.5095%CI [1.05,2.14], P=0.03) VEGF rs3025039共顯性模型-純和型／野生型(TT Vs CC)與DR發(fā)病風(fēng)險(xiǎn)存在相關(guān)性(I2=50%, OR=2.4795%CI [1.11,5.51], P=0.03),特別是在黃色人種中(I2=51%, OR=3.5395%CI [1.35,9.26], P=0.01)。隱性模型(TT Vs CT+CC)白色人種與DR發(fā)病無(wú)明顯相關(guān)性(P=0.05),在黃色人種中示與DR的發(fā)生相關(guān)(I2=33%, OR=3.1995%CI [1.21,8.42], P=0.01);其它各模型與DR的發(fā)病無(wú)明顯相關(guān)性(P0.05)。 (2)本省人群中DWR組與DR組VEGF基因多態(tài)性位點(diǎn)基因型頻率及等位基因頻率： VEGF基因rs699947位點(diǎn)：DR組AA基因型分布頻率為14.8%,DWR組為4.3%；CC基因型DR組為48.1%,DWR組為60.9%(P=0.04,0.05),其等位基因A頻率在DR組為33.3%,DWR組為21.7%(P=0.016,0.05)。攜帶rs699947AA基因型的糖尿病患者發(fā)生DR的危險(xiǎn)性較rs699947CC基因型及CA基因型攜帶者高3.83倍(OR=3.83)。 VEGF基因rs833061位點(diǎn)：DR組CC基因型分布頻率為16.0%,DWR組為4.3%；TT基因型DR組為45.7%,DWR組為58.7%(P=0.025,0.05),其等位基因C頻率在DR組為35.2%,DWR組為22.8%(P=0.011,0.05)。攜帶rs833061CC基因型的糖尿病患者發(fā)生DR的危險(xiǎn)性較rs833061TT基因型及TC基因型攜帶者高4.21倍(OR=4.21)。 VEGF基因rs13207351位點(diǎn)：基因型頻率及等位基因頻率在研究組和對(duì)照組間的差異無(wú)明顯統(tǒng)計(jì)學(xué)意義(P0.05)。攜帶rs13207351GG基因型、GA基因型及AA基因型的糖尿病患者發(fā)生DR的危險(xiǎn)性沒(méi)有明顯差別。 VEGF基因rs2010963位點(diǎn)：DR組CC基因型分布頻率為21.0%,DWR組為9.8%。GG基因型DR組為28.4%,DWR組為42.4%(P=0.049,0.05),其等位基因C頻率在DR組為46.3%,DWR組為33.4%(P=0.017,0.05)。攜帶rs2010963CC基因型的糖尿病患者發(fā)生DR的危險(xiǎn)性較rs2010963GG基因型及GC基因型攜帶者高2.45倍(OR=2.45)。 VEGF基因rs3025039位點(diǎn)：基因型頻率及等位基因頻率在研究組和對(duì)照組間的差異無(wú)明顯統(tǒng)計(jì)學(xué)意義(P0.05)。攜帶rs3025039CC基因型、CT基因型及TT基因型的糖尿病患者發(fā)生DR的概率沒(méi)有明顯統(tǒng)計(jì)差異。 (3) VEGF蛋白總量、VEGF165含量和VEGF165b含量： 1)正常對(duì)照組中血清VEGF總量為44.47±3.42(pg/ml), DWR組血清VEGF總量為57.74±11.23(pg/ml), DR組59.25±11.62(pg/ml),DWR組及DR組的VEGF總量都較正常對(duì)照組升高(P0.05),但DWR組與DR組之間VEGF總量沒(méi)有明顯的統(tǒng)計(jì)學(xué)差異(P0.05)。 2)正常對(duì)照組中VEGF165含量為15.89±1.31(pg/ml), DWR組為22.68±4.52(pg/ml); DR組為27.43±7.29(pg/ml),與正常對(duì)照組比具有統(tǒng)計(jì)學(xué)意義(P0.05)；且VEGF165在D WR組及DR組之間也有顯著差異(P=0.025)。 3)VEGF165b在血中的含量在正常對(duì)照組為9.25±0.82(pg/ml); DWR組為6.97±1.46(pg/ml); DR組為5.43±2.35(pg/ml).其含量在三組之間呈現(xiàn)逐漸降低的趨勢(shì),差異具有統(tǒng)計(jì)學(xué)意義(P0.05)。VEGF165及VEGF165b的比值在三組間出現(xiàn)變化,比值逐漸增大(1.72、3.25、5.05)。 4)蛋白總量與基因位點(diǎn)：攜帶rs2010963CC基因型人群其血中VEGF蛋白含量較GG基因型及GA型顯著增高(CC基因型VEGF蛋白含量為62.59±13.71(pg/ml),GA基因型蛋白含量為53.27±10.37(pg/ml),GG基因型蛋白含量為50.85±10.59(pg/ml));CC基因型與GG基因型及GC基因型比較均具有統(tǒng)計(jì)學(xué)上的顯著性差異,P0.05),其他各SNP位點(diǎn)各基因型之間無(wú)明顯統(tǒng)計(jì)學(xué)差別(P0.05)。 (4)糖尿病視網(wǎng)膜病變易感因素分析： 1)DWR組及DR組之間的人口學(xué)特征及生化指標(biāo)的相關(guān)數(shù)據(jù)進(jìn)行成組T檢驗(yàn),發(fā)現(xiàn)收縮壓、舒張壓、糖化血紅蛋白及肌酐值在兩組之間存在顯著的統(tǒng)計(jì)學(xué)差異(P0.05)。 2)logistic回歸分析：收縮壓高于160mmHg的糖尿病患者發(fā)生DR的危險(xiǎn)性較低于130mmHg的高7.5倍,舒張壓高于90mmHg的糖尿病患者發(fā)生DR的危險(xiǎn)性較低于90mmHg的高2倍。HbAIC含量高于14.0%的糖尿病患者發(fā)生DR的危險(xiǎn)性較低于8.0%的高11.50倍。肌酐值高于100mmol/L的糖尿病患者發(fā)生DR的危險(xiǎn)性較低于60mmol/L的高出5.5倍。攜帶rs2010963CC基因型糖尿病患者發(fā)生DR的危險(xiǎn)性較GG基因型患者高3.2倍。VEGF165含量高于28.0mmol/L的糖尿病患者發(fā)生DR的危險(xiǎn)性較低于28.0mmol/L的患者高9.3倍,而VEGF165b含量高于5.0mmol/L的糖尿病患者發(fā)生DR的危險(xiǎn)性較低于5.0mmol/L的患者明顯降低(OR=0.013),VEGF165b對(duì)于糖尿病患者有明顯保護(hù)作用。 結(jié)論： (1)黃色人種中VEGF rs699947的共顯性模型(C A Vs C C)、 VEGF rs2010963隱性模型(CC Vs GC+GG)及VEGF rs3025039的共顯性模型(TT Vs CC)、隱性模型(TT Vs CT+CC)跟糖尿病視網(wǎng)膜病變的發(fā)生相關(guān)；而在白色人種中上述多態(tài)性位點(diǎn)與糖尿病視網(wǎng)膜病變無(wú)明顯相關(guān)性。 (2)本省人群VEGF基因rs699947位點(diǎn)、VEGF基因rs833061位點(diǎn)、VEGF基因rs2010963位點(diǎn)的多態(tài)性變化在糖尿病視網(wǎng)膜病變發(fā)生發(fā)展中具有一定的意義 (3)VEGFXXX與EGFxxxb比值改變導(dǎo)致糖尿病視網(wǎng)膜病變的發(fā)生和發(fā)展。rs2010963位點(diǎn)多態(tài)性變化為功能性改變,對(duì)糖尿病患者發(fā)生糖尿病視網(wǎng)膜病變有重要意義。 (4)收縮壓≥160mmHg舒張壓≥90mmHg.糖化血紅蛋白≥8.0mmol/L、肌酐值≥VEGF16528mmol/L、rs2010963CC基因型及VEGF165b5.0mmol/L為糖尿病視網(wǎng)膜病變發(fā)生的高危人群。
[Abstract]:Objective:
Objective To study the genetic and environmental susceptibility factors of type 2 diabetic retinopathy (DR) in Hunan province, and to screen the high risk population for DR.
(1) Single nucleotide polymorphisms of vascular endothelial growth factor (VEGF) associated with diabetic retinopathy in yellow race were searched by meta-analysis.
(2) To explore the association between single nucleotide polymorphism of vascular endothelial growth factor (VEGF) and susceptibility to diabetic retinopathy in Hunan population.
(3) To investigate the correlation between serum total VEGF protein, subtype VEGF 165 and its isomer VEGF 165b and diabetic retinopathy.
(4) Case-control and logistic regression analysis were used to compare the single nucleotide polymorphisms of vascular endothelial growth factor (VEGF), the protein of vascular endothelial growth factor (VEGF) and other demographic and biochemical characteristics, and to explore the risk factors of diabetic retinopathy.
Method:
(1) To review all the English literature on the relationship between single nucleotide polymorphism of vascular endothelial growth factor and diabetic retinopathy before March 2012.
(2) According to meta-analysis, the significant single nucleotide polymorphisms of vascular endothelial growth factor (VEGF) in yellow race were detected by restriction fragment length polymorphism polymerase chain reaction (PCR-RFLR) in Hunan, diabetic without retinopathy (DWR), diabetic retinopathy (DR) and diabetic retinopathy (DR). The distribution of diabetic retinopathy (DR) was analyzed by chi-square test.
(3) Serum levels of VEGF, subtype/EGF165 and its isomer, VEGF 165b, were measured by enzyme-linked immunosorbent assay (ELISA) in Hunan normal subjects, diabetic patients without retinopathy and diabetic patients with retinopathy. The correlation between /EGF and single nucleotide polymorphisms and environmental factors.
(4) The demographic characteristics and biochemical data of diabetic retinopathy patients without retinopathy and diabetic retinopathy patients with diabetic retinopathy and the expression of VEGF single nucleotide polymorphism site and VEGF protein were analyzed by case-control method. T-test and logistic regression were used to explore the risk factors of diabetic retinopathy.
Result:
(1) meta analysis results:
Vascular growth factor rs699947 co-dominant model-heterozygous/wild type (CAVs CC) was associated with DR risk (I 2 = 64%, OR = 1.2795% CI [1.05, 1.54], P = 0.02), especially in yellow people (I 2 = 38%, OR = 1.4695% CI [1.17, 1.82], P = 0.0007). Other models had no significant correlation with DR risk (P 0.05).
There was no significant correlation between the VEGF rs1570360 models and the incidence of DR in all races (P0.05).
Subgroup analysis of VEGF rs2010963: Recessive model of yellow race group (CC V s GC+GG) showed a significant association with DR (I 2 = 60%, OR = 1.2895% CI [1.01, 1.63], P = 0.04); recessive model of PCR-RFLR sequencing group (CC V s GC+GG) was significantly associated with DR (I 2 = 46%, OR = 1.5095% CI [1.05, 2.14], P = 0.03)
The co-dominant model of VEGF rs3025039-pure and wild type (TT Vs CC) was correlated with the risk of DR (I 2 = 50%, OR = 2.4795% CI [1.11, 5.51], P = 0.03), especially in yellow races (I 2 = 51%, OR = 3.5395% CI [1.35, 9.26], P = 0.01). The recessive model (TT Vs CT + CC) had no significant correlation with DR (P = 0.05) in white races and yellow races (P = 0.05). It was found that DR was associated with DR (I 2 = 33%, OR = 3.1995% CI [1.21, 8.42], P = 0.01); other models had no significant correlation with DR (P 0.05).
(2) genotype frequencies and allele frequencies of VEGF gene polymorphisms in DWR and DR groups in this province:
The distribution frequency of vascular endothelial growth factor gene rs699947 was 14.8% in DR group and 4.3% in DWR group, 48.1% in CC genotype DR group, 60.9% in DWR group (P = 0.04, 0.05). The allele A frequency was 33.3% in DR group and 21.7% in DWR group (P = 0.016, 0.05). The risk of DR in diabetic patients with rs699947AA genotype was higher than that of rs699947CC genotype and CA gene. The carriers were 3.83 times higher (OR=3.83).
The distribution frequency of vascular endothelial growth factor gene rs833061 was 16.0% in DR group and 4.3% in DWR group, 45.7% in TT genotype DR group and 58.7% in DWR group (P = 0.025, 0.05). The allele C frequency was 35.2% in DR group and 22.8% in DWR group (P = 0.011, 0.05). The risk of DR in diabetic patients with rs833061CC genotype was higher than that of rs833061TT genotype and TC gene. The carriers were 4.21 times higher (OR=4.21).
There was no significant difference in genotype frequency and allele frequency between the study group and the control group (P 0.05). There was no significant difference in the risk of DR between diabetic patients with rs13207351GG genotype, GA genotype and AA genotype.
The distribution frequency of CC genotype was 21.0% in DR group and 9.8% in DWR group, 28.4% in GG genotype DR group, 42.4% in DWR group (P = 0.049, 0.05), and 46.3% in DR group and 33.4% in DWR group. The carriers were 2.45 times higher (OR=2.45).
There was no significant difference in genotype frequency and allele frequency between the study group and the control group (P 0.05). There was no significant difference in the incidence of DR among diabetic patients with rs3025039CC genotype, CT genotype and TT genotype.
(3) total VEGF protein, VEGF165 content and VEGF165b content:
1) The total level of serum VEGF in normal control group was 44.47 (+ 3.42) (pg / ml), 57.74 (+ 11.23) (pg / ml) in DWR group, 59.25 (+ 11.62) (pg / ml) in DR group, and increased in DWR group and DR group compared with normal control group (P 0.05), but there was no significant difference between DWR group and DR group (P 0.05).
2) The content of VEGF 165 in normal control group was 15.89 (+ 1.31) (pg/ml), 22.68 (+ 4.52) (pg/ml) in DWR group, 27.43 (+ 7.29) (pg/ml) in DR group, and there was significant difference between DWR group and DR group (P = 0.025).
3) The levels of VEGF 165b in blood were 9.25 (+ 0.82) (pg/ml) in the normal control group, 6.97 (+ 1.46) (pg/ml) in the DWR group, and 5.43 (+ 2.35) (pg/ml) in the DR group. The levels of VEGF 165b decreased gradually among the three groups, and the difference was statistically significant (P 0.05). The ratios of VEGF 165 and VEGF 165b changed among the three groups, and gradually increased (1.72, 3.25, 5.05).
4) Total protein and gene locus: The content of VEGF protein in serum of the population with rs2010963CC genotype was significantly higher than that of GG genotype and GA genotype (the content of VEGF protein in CC genotype was 62.59 [13.71], that of GA genotype was 53.27 [10.37], and that of GG genotype was 50.85 [10.59], that of CC genotype and GG genotype and GC genotype were significantly higher than that of the population with rs2010963CC genotype and GA genotype. Genotype comparison showed significant difference (P 0.05), and there was no significant difference among other SNP loci (P 0.05).
(4) predisposing factors of diabetic retinopathy:
1) The data of demographic characteristics and biochemical indexes between DWR group and DR group were tested by T-test in groups. The results showed that there were significant differences in systolic blood pressure, diastolic blood pressure, glycosylated hemoglobin and creatinine between the two groups (P 0.05).
2) Logistic regression analysis: diabetic patients with systolic blood pressure higher than 160 mmHg had a 7.5-fold lower risk of DR than 130 mmHg, diabetic patients with diastolic blood pressure higher than 90 mmHg had a 2-fold higher risk of DR than 90 mmHg, diabetic patients with HbAIC content higher than 14.0% had a 11.50-fold higher risk of DR than 8.0% and high creatinine value. The risk of DR was 5.5 times lower in diabetics with 100 mmol/L than in patients with 60 mmol/L. The risk of DR was 3.2 times higher in diabetics with rs2010963CC gene than in patients with GG genotype. The risk of DR in 5.0 mmol/L diabetic patients was significantly lower than that in 5.0 mmol/L diabetic patients (OR = 0.013). VEGF 165b has a protective effect on diabetic patients.
Conclusion:
(1) The co-dominant model (C A Vs C), recessive model (CC Vs GC+GG) and co-dominant model (TT Vs CC) of VEGF rs699947 and recessive model (TT Vs CT+CC) of VEGF rs3025039 were associated with diabetic retinopathy in yellow people, but the above polymorphisms were not associated with diabetic retinopathy in white people. Customs.
(2) The polymorphism of rs699947, rs833061 and rs2010963 of vascular endothelial growth factor gene may play an important role in the development of diabetic retinopathy.
(3) The change of the ratio of VEGF XXX to EGF xxxb leads to the occurrence and development of diabetic retinopathy. The change of rs2010963 polymorphism is a functional change, which is important for diabetic retinopathy.
(4) Systolic blood pressure (> 160mmHg) diastolic blood pressure (> 90mmHg), glycosylated hemoglobin (> 8.0mmol/L), creatinine (> VEGF 16528mmol/L), rs2010963CC genotype and VEGF 165b5.0mmol/L are high risk groups for diabetic retinopathy.
【學(xué)位授予單位】：中南大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2013
【分類(lèi)號(hào)】：R587.2;R774.1

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相關(guān)期刊論文 前2條

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2 李清解,謝平,黃建軍,谷亞鵬,曾衛(wèi)民,宋惠萍;中國(guó)人2型糖尿病患者醛糖還原酶基因5調(diào)控區(qū)的多態(tài)性與功能(英文)[J];Chinese Medical Journal;2002年02期

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