本文選題：糖尿病視網(wǎng)膜病變 + 脈絡(luò)膜��； 參考：《天津醫(yī)科大學(xué)》2014年博士論文

【摘要】：目的：應(yīng)用深度增強成像技術(shù)研究不同階段的糖尿病視網(wǎng)膜病變(diabetic retinopathy, DR)和糖尿病黃斑水腫(diabetic macular edema, DME)黃斑區(qū)脈絡(luò)膜厚度變化,建立視網(wǎng)膜muller細胞原代培養(yǎng)體系,探討不同階段DR與PPARy基因單核苷酸多態(tài)性的相關(guān)性及其影響因素。 方法：選擇正常志愿者共120例,年齡25～85歲。根據(jù)研究對象不同年齡分組。深度增強成像相干光斷層掃描測量脈絡(luò)膜厚度。分析黃斑中心凹脈絡(luò)膜厚度(subfoveal choroidal thickness, SFCT)與年齡的相關(guān)性。收集2型糖尿病患者92例(92眼),共分為3組：A組(DR-/DME-):無DR無DME組36例(36眼)；B組(NPDR+/DME-):無DME的非增生性糖尿病視網(wǎng)膜病變(Non-proliferative diabetic retinopathy, NPDR)組47例(47眼)；C組(PDR+/DME-):無DME的增生性糖尿病視網(wǎng)膜病變(Proliferative diabetic retinopathy, PDR)組9例(9眼),分別比較組間脈絡(luò)膜厚度的差異、各組與正常人群脈絡(luò)膜厚度的差異。選擇DME33例33眼,采集指標：空腹血糖、糖化血紅蛋白(Glycosylated hemoglobin, HbAlc)、總膽固醇(Total cholesterol, TC)、甘油三酯(Triglycerides, TG)、高密度脂蛋白(High-density lipoprotein, HDL)、低密度脂蛋白(Low-density lipoprotein, LDL)等。比較DME與正常人群及不同階段DR的脈絡(luò)膜厚度。分析糖尿病病程、空腹血糖、HbAlc、TG、TC、LDL、HDL、UAER等因素與DME脈絡(luò)膜厚度變化的相關(guān)性。另選擇不同階段DR120例120眼,直接測序法檢測PPARγpro12Ala位點基因型,比較不同階段DR等位基因和基因型頻率分布差異,分析等位基因和基因型與DR發(fā)生的相關(guān)性；分析PPARy基因pro12Ala位點多態(tài)性與不同階段DR的相關(guān)性。進一步探討不同等位基因與TG、LDL、HDL、HbAlc、UAER等臨床生化指標的相關(guān)性。同時應(yīng)用改良方法進行體外原代培養(yǎng)并純化鑒定SD大鼠視網(wǎng)膜muller細胞,為下一步研究提供實驗基礎(chǔ)。 結(jié)果：正常人群平均SFCT為(271.32±35.63)μm男性SFCT平均值為(286.32±35.98)μm,60名女性為(254.33±39.61)μm,男性平均SFCT高于女性,且兩者之間差異有統(tǒng)計學(xué)意義(P0.05)。不同年齡組之間SFCT差異有統(tǒng)計學(xué)意義(P0.001)。隨著年齡的增加,SFCT逐漸減小,與年齡呈負相關(guān)(r=-0.781,P0.001)。A組：(DR-/DME-)的SFCT為145-306μm,平均為(271.19±34.68)μm,與正常對照人群的平均SFCT進行比較,差異無統(tǒng)計學(xué)意義(P0.05)。B組：(NPDR+/DME-)的SFCT為176～336μm,平均為(259.64±37.79)μm,與正常對照的平均SFCT比較差異無統(tǒng)計學(xué)意義(P0.05)。C明：(PDR+/DME-)的SFCT為171～289μm,平均(232.98±34.13)μm,與正常對照的平均SFCT比較,脈絡(luò)膜明顯變薄,且差異具有統(tǒng)計學(xué)意義(P0.05)。D組：(NPDR+/DME+)的SFCT為156～278μm,平均為(229.64±33.66)μm,與正常對照45～65歲(含)年齡組的平均SFCT(270.78±35.97)μm比較,脈絡(luò)膜明混變薄,差異具有顯著統(tǒng)計學(xué)意義(P0.01)。D組與B組及正常人群比較,D組脈絡(luò)膜明顯變薄,且差異具有統(tǒng)計學(xué)意義(P0.01)。HbAlc、LDL、UAER水平是DME發(fā)生的危險因素,其Exp(B)分別是5.512(P0.001)；2.821(P=0.017)；1.582(P=0.009)。合并DME的NPDR脈絡(luò)膜厚度與LDL和UAER具有相關(guān)性(r=-0.601、-0.673,P0.01),而與糖尿病病程、空腹血糖、HbAlc TG、TC、HDL、肌酐、收縮壓及舒張壓均無相關(guān)性(r-=-0.253、0.219、0.095、0.310、0.185、0.224、0.237、0.421、0.281,P0.05)。通過胰蛋白酶多次反復(fù)短時間消化,以谷氨酰胺合成酶為特異性抗體進行細胞鑒定,體外培養(yǎng)的muller細胞形態(tài)一致,谷氨酰胺合成酶反應(yīng)陽性。DR患者PPARy基因G等位基因的分布頻率為5.8%,PDR和NPDR患者基因型均以C/C型為主導(dǎo),所占比例分別為87%和89.2%,二者之間差異無統(tǒng)計學(xué)意義。不同基因型全身主要生化指標比較顯示：與CC基因型的DR患者相比,G/G或G/C基因型患者UAER水平明顯降低,具有統(tǒng)計學(xué)差異,而TG、LDL、HDL、HbAlc水平差異無統(tǒng)計學(xué)意義。結(jié)論：正常中國人平均SFCT為271.32±35.63gm,各年齡段男性平均SFCT高于女性。隨著年齡的增加,SFCT逐漸減小,與年齡呈負相關(guān)。鼻側(cè)脈絡(luò)膜均最薄。隨著DR加重,脈絡(luò)膜厚度呈降低趨勢。無DR的PDR患者脈絡(luò)膜較正常人及無DR糖尿病患者均明顯變薄,與無DME的NPDR目比,合并DME的NPDR脈絡(luò)膜顯著變薄。LDL和UAER水平與DME脈絡(luò)膜厚度呈負相關(guān)。胰蛋白酶反復(fù)消化培養(yǎng)是一種可靠的獲得較為純化、表達穩(wěn)定的muller細胞體外原代培養(yǎng)的方法。PPARy基因pro12Ala位點的基因型和等位基因的分布頻率與DR的發(fā)生,以及不同階段的DR無明確相關(guān)性；但PPARγ基因G等位基因可能減低DR患者UAER值,對其腎臟功能可能具有一定保護作用。
[Abstract]:Objective: To study the changes of the choroidal thickness in the macular region of diabetic retinopathy (diabetic retinopathy, DR) and diabetic macular edema (diabetic macular edema, DME) by depth enhanced imaging, and to establish a primary culture system for retinal Muller cells and explore the single nucleotide polymorphisms of DR and PPARy genes at different stages. Correlation and its influencing factors.
Methods: a total of 120 normal volunteers, aged 25~85 years, were divided into different age groups. The depth enhanced imaging coherent light tomography was used to measure choroidal thickness. The correlation between subfoveal choroidal thickness (SFCT) and age was analyzed. 92 cases of type 2 diabetes mellitus (92 eyes) were collected, which were divided into 3. Group A (DR-/DME-): no DR without DME group 36 cases (36 eyes); B group (NPDR+/DME-): non proliferative diabetic retinopathy of DME (Non-proliferative diabetic retinopathy, NPDR) group 47 cases (47 eyes); C group: 9 cases (9 eyes) of proliferative diabetic retinopathy (9 eyes), respectively. The difference of choroidal thickness between the groups and the choroidal thickness of the normal group. Select DME33 33 eyes and collect the indexes: fasting blood glucose, glycated hemoglobin (Glycosylated hemoglobin, HbAlc), total cholesterol (Total cholesterol, TC), triglyceride (Triglycerides, TG), high density lipoprotein (High-density lipoprotein, HDL), Low density lipoprotein (Low-density lipoprotein, LDL) and so on. Compare the choroidal thickness of DME and normal people and different stages of DR. Analysis of the correlation between diabetes course, fasting blood glucose, HbAlc, TG, TC, LDL, HDL, UAER and other factors with the changes of the thickness of the DME choroid. The other 120 eyes were selected at different stages and direct sequencing was used to detect the gamma locus Genotypes, compare the difference of DR allele and genotype frequency distribution at different stages, analyze the correlation between alleles and genotypes and DR, analyze the correlation between pro12Ala polymorphism of PPARy gene and DR at different stages, and further discuss the correlation of different alleles with TG, LDL, HDL, HbAlc, UAER and other clinical biochemical indexes. The Muller cells of SD rat retina were cultured and purified in vitro by improved method, providing experimental basis for further research.
Results: the average SFCT of the normal population was (271.32 + 35.63) mu m male SFCT mean (286.32 + 35.98) mu m, 60 women (254.33 + 39.61) m, and the average male SFCT was higher than the female, and the difference between the two groups was statistically significant (P0.05). The SFCT difference between different age groups was statistically significant (P0.001). As the age increased, SFCT gradually decreased, The negative correlation with age (r=-0.781, P0.001) group.A: (DR-/DME-) SFCT was 145-306 mu m, the average was (271.19 + 34.68) mu m, compared with the average SFCT in the normal control group, the difference was not statistically significant (P0.05).B group: (NPDR+/DME-) SFCT was 176~336 mu, the average was (259.64 + 37.79) micron, and there was no difference compared with the normal control. Statistical significance (P0.05).C clear: (PDR+/DME-) SFCT is 171~289 mu m, average (232.98 + 34.13) mu m, compared with the average SFCT of normal control, the choroid membrane is thinner, and the difference has statistical significance (P0.05).D group: (NPDR+/DME+) SFCT 156~278 mu m, average (229.64 + 33.66) mu, and normal control 45~65 years (containing) age group flat level Compared with SFCT (270.78 + 35.97) mu m, choroidal Ming was thinner and thinner, the difference had significant statistical significance (P0.01), compared with the B group and the normal group, the choroidal membrane in the D group was obviously thinner, and the difference was statistically significant (P0.01).HbAlc, LDL, UAER level was the risk factor of DME, and the Exp (2.821); 1.582 (1.582) 0.009. The NPDR choroid thickness of the combined DME was associated with LDL and UAER (r=-0.601, -0.673, P0.01), but had no correlation with the course of diabetes, fasting blood glucose, HbAlc TG, TC, HDL, creatinine, systolic and diastolic pressure. Muller cells were identified by glutamine synthetase for specific antibodies, and the morphology of the cultured Muller cells in vitro was the same. The frequency of PPARy gene G alleles in.DR patients with glutamine synthetase positive reaction was 5.8%. The genotype of PDR and NPDR patients were dominated by C/C type, and the proportion was 87% and 89.2% respectively. There was no statistical difference between the two groups. Comparison of the main biochemical indexes of different genotypes showed that compared with the DR patients with CC genotype, the UAER level of the patients with G/G or G/C genotype decreased significantly, with statistical difference, but there was no statistical difference between TG, LDL, HDL and HbAlc. Conclusion: the average SFCT of normal Chinese was 271.32 + 35.63gm, and the average SFCT was higher in men of all ages. With the increase of age, SFCT gradually decreased and had a negative correlation with age. The nasal choroid was thinnest. With the increase of DR, the choroidal thickness decreased. The choroid of the PDR patients without DR was significantly thinner than the normal and non DR diabetics, compared with the DME free NPDR mesh, the NPDR choroid with DME's NPDR choroid was significantly thinner.LDL and UAER and DME. Choroidal thickness is negative correlation. Repeated trypsin digestion and culture is a reliable, purified, stable Muller cell in vitro primary culture. The genotype and allele distribution frequency of the.PPARy gene pro12Ala locus and the occurrence of DR, as well as the DR in different stages; but G alleles of the PPAR gamma gene. The gene may reduce the UAER value of DR patients, and may have some protective effect on renal function.
【學(xué)位授予單位】：天津醫(yī)科大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2014
【分類號】：R587.2;R774.1

【參考文獻】

中國期刊全文數(shù)據(jù)庫 前6條

1 Amelah Mohamed Abdul Qader;Zunaina Embong;Bakiah Shaharuddin;Shahrul Bariyah Sahul Hamid;Shatriah Ismail;;血清血脂分析結(jié)果與2型糖尿病患者視網(wǎng)膜硬性滲出的關(guān)系(英文)[J];國際眼科雜志;2009年04期

2 茍琳,張作明,許漢鵬,吳玉梅,晏楠,郭守一;恒溫搖動法純化培養(yǎng)大鼠視網(wǎng)膜Müller細胞的技術(shù)研究[J];眼科研究;2002年05期

3 彭廣華,李志杰,李辰;視網(wǎng)膜Müller細胞原代培養(yǎng)的技術(shù)研究[J];眼科研究;1998年01期

4 康奕飛;何兆初;吳竣;曾昭華;;PPAR-α激動劑對大鼠內(nèi)皮功能不全的作用及機制探討[J];醫(yī)學(xué)理論與實踐;2008年06期

5 魏楓;王津京;王彩麗;霍曉靜;薛曄霞;白琳院;;2型糖尿病腎病與PPAR-γ2 Pro12Ala單核苷酸多態(tài)性的相關(guān)性研究[J];中國糖尿病雜志;2008年11期

6 胡竹林,杜蜀華;壓力對大鼠純化視網(wǎng)膜神經(jīng)節(jié)細胞誘導(dǎo)型一氧化氮合酶mRNA及其蛋白表達的影響[J];中華眼科雜志;2002年08期

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