本文選題：糖尿病性視網(wǎng)膜病變 + 促紅細胞生成素��； 參考：《武漢大學》2011年博士論文

【摘要】：糖尿病性視網(wǎng)膜病變是以視網(wǎng)膜新生血管性增殖為特征的一種致盲性眼病,隨著人們生活水平的提高,糖尿病日益成為我國威脅人類健康的常見多發(fā)病,與此相應,糖尿病性視網(wǎng)膜病變的發(fā)病也日益增多。迄今為止,糖尿病性視網(wǎng)膜病變的確切發(fā)病機制還不完全清楚,大量的研究認為視網(wǎng)膜新生血管形成的過程與酶、激酶的調(diào)節(jié)以及各種生長因子的作用有關(guān),同時炎癥及免疫反應也在此過程中扮演著重要的作用。 第一部分增殖性糖尿病性視網(wǎng)膜病變患者血清及玻璃體樣本中GRO-a及EPO的表達 [目的]通過檢測糖尿病性視網(wǎng)膜病變(DR)患者的血漿、玻璃體樣本內(nèi)促紅細胞生成素(EPO)、生長相關(guān)基因α(Gro-a)的含量,探討其在DR發(fā)展變化中的可能作用。 [方法]該研究為病例對照研究。包括非增生型糖尿病性視網(wǎng)膜病變(NPDR)組、增生型糖尿病性視網(wǎng)膜病變(PDR)組與對照組。采用酶聯(lián)免疫吸附試驗(ELISA)檢測各組血漿、玻璃體樣本中EPO和Gro-a的含量。 [結(jié)果]血漿EPO含量：NPDR組(22.16±4.85) mIU/ml, PDR組(25.46±8.83)mIU/ml,對照組(23.52±7.27)mIU/ml。玻璃體EPO含量：PDR組(461.36±101.20)mIU/ml,對照組(36.78±10.19)mIU/ml。血漿Gro-a含量：NPDR組(81.95±38.08)pg/ml, PDR組(82.61±25.26)pg/ml,對照組(55.68±22.53) pg/ml。玻璃體Gro-a含量：PDR組(327.74±216.29) pg/ml,對照組(81.95±47.59)pg/ml。3組血漿樣本間EPO含量差異無統(tǒng)計學意義(P0.05), Gro-a含量差異有統(tǒng)計學意義(P0.05)。3組玻璃體樣本內(nèi)PDR組與對照組2組間EPO、Gro-α差異均有顯著統(tǒng)計學意義(P0.05)。各組血漿、玻璃體內(nèi)EPO、Gro-α含量與相應糖化血紅蛋白(HbA1c%)有不同程度正相關(guān)關(guān)系。 [結(jié)論]PDR患者血漿Gro-a含量,玻璃體EPO、Gro-a含量顯著增高,EPO、Gro-a含量與患者HbA1c (%)值有關(guān)。 第二部分GRO-a, EPO在糖尿病大鼠視網(wǎng)膜組織中的表達及塞來昔布 對GRO-α、EPO表達的影響 [目的]觀察GRO-a及EPO在糖尿病大鼠視網(wǎng)膜組織中的表達,以及選擇性環(huán)氧合酶-2 (Cyclooxygenase-2, COX-2)抑制劑(塞來昔布)對糖尿病大鼠視網(wǎng)膜組織中GRO-α和EPO表達的影響。 [方法]將40只SD大鼠隨機分為4組,即正常組、糖尿病組、糖尿病+蒸餾水灌胃組以及糖尿病+塞來昔布灌胃組,每組10只。通過腹腔注射鏈脲佐菌素(streptoz-otocin, STZ)誘導建立糖尿病大鼠動物模型,飼養(yǎng)12周后處死大鼠,取視網(wǎng)膜,SYBR熒光實時定量PCR檢測視網(wǎng)膜組織中COX-2、血管內(nèi)皮生長因子(vascular endothelial growth factor, VEGF)、EPO和GRO-a mRNA的表達變化,western-blotting檢測大鼠視網(wǎng)膜中COX-2、VEGF、EPO和GRO-a蛋白的表達變化。通過Quantity One凝膠圖像分析軟件進行結(jié)果分析,行ANOVA方差分析,SNK法做組間兩兩比較,以P0.05作為差異有統(tǒng)計學意義。 [結(jié)果]12周后,COX-2, VEGF, EPO及GRO-a mRNA和蛋白質(zhì)的表達在糖尿病組和糖尿病+蒸餾水組最高,糖尿病+塞來昔布灌胃組高于正常組而低于糖尿病組。糖尿病組和糖尿病+蒸餾水組間差異無統(tǒng)計學意義(P0.05),其余各組間比較均有顯著性差異(P0.05)。 [結(jié)論]EPO和GRO-a在糖尿病大鼠視網(wǎng)膜新生血管生成中起到了重要作用,塞來昔布可能可以通過抑制糖尿病大鼠視網(wǎng)膜中EPO和GRO-a的表達,從而抑制糖尿病大鼠視網(wǎng)膜組織中VEGF的表達,最終起到抗新生血管形成的作用。 第三部分塞來昔布對糖尿病大鼠視網(wǎng)膜組織中d114、Notch-1表達的影響 [目的]觀察選擇性環(huán)氧合酶-2 (Cyclooxygenase-2,COX-2)抑制劑(塞來昔布)對糖尿病大鼠視網(wǎng)膜組織中Delta樣配體4- Notch信號(delta-like 4 ligan-Notch, D114/Notch-1)通路的影響。 [方法]將40只SD大鼠隨機分為4組,即正常組、糖尿病組、糖尿病+蒸餾水灌胃組以及糖尿病+塞來昔布灌胃組,每組10只。通過腹腔注射鏈脲佐菌素(streptoz-otocin,STZ)誘導建立糖尿病動物模型,飼養(yǎng)12周后處死大鼠,取視網(wǎng)膜組織,SYBR熒光實時定量PCR檢測視網(wǎng)膜組織中COX-2、血管內(nèi)皮生長因子(vascular endothelial growth factor,VEGF)以及D114/Notch-1 mRNA的表達變化,western-blotting檢測大鼠視網(wǎng)膜組織中COX-2、VEGF以及D114/Notch-1蛋白的表達。通過Quantity One凝膠圖像分析軟件進行結(jié)果分析,行ANOVA方差分析,SNK法做組間兩兩比較,以P0.05作為差異有統(tǒng)計學意義。 [結(jié)果]12周后,COX-2、VEGF、D114及Notch-1 mRNA和蛋白質(zhì)的表達在糖尿病組和糖尿病+蒸餾水組間差異無統(tǒng)計學意義(P=0.121),其余各組間比較均有顯著性差異(P0.05)。正常組表達最少,糖尿病組和糖尿病+蒸餾水組表達最高,COX-2、VEGF mRNA和蛋白質(zhì)表達在塞來昔布用藥組高于正常組,但低于糖尿病組以及糖尿病+蒸餾水組；而D114/Notch-1 mRNA和蛋白質(zhì)在塞來昔布用藥組表達最高。 [結(jié)論]D114和Notch-1在糖尿病大鼠視網(wǎng)膜的新生血管生成過程中起到了重要作用,選擇性COX-2抑制劑塞來昔布可以通過激活糖尿病大鼠視網(wǎng)膜中的D114/ Notch-1表達,從而抑制糖尿病大鼠視網(wǎng)膜組織中的VEGF表達。
[Abstract]:Diabetic retinopathy is a kind of blinding eye disease characterized by retinal neovascularization. With the improvement of people's living standard, diabetes is becoming a common and frequently occurring disease which threatens human health in our country. Accordingly, the incidence of diabetic retinopathy is increasing. So far, diabetic retinopathy The exact pathogenesis of the change is not completely clear. A large number of studies suggest that the formation of retinal neovascularization is related to the regulation of enzymes and kinases and the effects of various growth factors, while inflammation and immune responses also play an important role in this process.
Part 1 expression of GRO-a and EPO in serum and vitreous samples from patients with proliferative diabetic retinopathy
[Objective] to investigate the possible role of erythropoietin (EPO) and growth related gene alpha (Gro-a) in the plasma of patients with diabetic retinopathy (DR) in the development and changes of DR.
[Methods] the study was a case-control study, including non proliferative diabetic retinopathy (NPDR) group, proliferative diabetic retinopathy (PDR) group and control group. The levels of plasma, EPO and Gro-a in the vitreous samples were detected by enzyme linked immunosorbent assay (ELISA).
[results] the plasma EPO content: NPDR group (22.16 + 4.85) mIU/ml, PDR group (25.46 + 8.83) mIU/ml, control group (23.52 + 7.27) mIU/ml., EPO content of vitreous body: PDR group (461.36 + 101.20) mIU/ml, and control group (36.78 + 10.19) mIU/ml. plasma Gro-a content: NPDR group (81.95 + 38.08) The content of body Gro-a: PDR group (327.74 + 216.29) pg/ml, and there was no significant difference between the plasma samples of the control group (81.95 + 47.59) pg/ml.3 (P0.05), and the difference of Gro-a content was statistically significant (P0.05) in the.3 group, the PDR group and the control group 2 groups were EPO, and the Gro- alpha difference was significant statistically significant. There was a positive correlation between EPO and Gro- alpha levels and corresponding glycosylated hemoglobin (HbA1c%) levels in vivo.
[Conclusion plasma levels of Gro-a and vitreous EPO and Gro-a in patients with]PDR were significantly increased, and the contents of EPO and Gro-a were related to HbA1c (%) value of patients.
The second part is the expression of GRO-a and EPO in retina of diabetic rats and celecoxib.
Influence on the expression of GRO- alpha and EPO
[Objective] to observe the expression of GRO-a and EPO in the retinal tissue of diabetic rats and the effect of selective cyclooxygenase -2 (Cyclooxygenase-2, COX-2) inhibitor (celecoxib) on the expression of GRO- alpha and EPO in the retina of diabetic rats.
[Methods] 40 SD rats were randomly divided into 4 groups: normal group, diabetic group, diabetes + distilled water gavage group and diabetic + celecoxib gavage group, 10 rats in each group were induced by intraperitoneal injection of streptozotocin (streptoz-otocin, STZ) to establish diabetic rat model. After 12 weeks of feeding, rats were killed, retina was taken and SYBR fluorescence was determined in real time. The expression of COX-2, vascular endothelial growth factor (vascular endothelial growth factor, VEGF), EPO and GRO-a mRNA in the retina of the retina was measured by PCR. Analysis, SNK method to do 22 comparisons between groups, P0.05 as the difference was statistically significant.
[results after]12 weeks, the expression of COX-2, VEGF, EPO and GRO-a mRNA and protein were highest in the diabetic group and the diabetic + distilled water group. The diabetes + celecoxib group was higher than the normal group but lower than the diabetic group. There was no significant difference between the diabetic group and the diabetes + distilled water group (P0.05), and the other groups had significant differences (P0). .05).
[conclusion]EPO and GRO-a play an important role in the formation of retinal neovascularization in diabetic rats. Celecoxib may inhibit the expression of EPO and GRO-a in the retina of diabetic rats and inhibit the expression of VEGF in the retina tissue of diabetic rats and eventually play a role in anti angiogenesis.
The third part is the effect of celecoxib on the expression of D114 and Notch-1 in retina of diabetic rats.
[Objective] to observe the effect of selective cyclooxygenase -2 (Cyclooxygenase-2, COX-2) inhibitor (celecoxib) on the Delta like ligand 4- Notch signal (Delta-like 4 ligan-Notch, D114/Notch-1) pathway in the retinal tissue of diabetic rats.
[Methods] 40 SD rats were randomly divided into 4 groups: normal group, diabetic group, diabetes + distilled water gavage group and diabetic + celecoxib gavage group, 10 rats in each group were induced by intraperitoneal injection of streptozotocin (streptoz-otocin, STZ) to establish diabetic animal model. After 12 weeks of feeding, rats were killed, retina tissue was taken and SYBR fluorescence was determined in real time. COX-2, vascular endothelial growth factor (VEGF) and the expression of D114/Notch-1 mRNA in retinal tissue were measured by PCR. Western-blotting was used to detect the COX-2, VEGF, and protein expression in the retina tissue of rats. Analysis of variance, SNK method made 22 comparisons between groups, P0.05 as the difference was statistically significant.
[results after]12 weeks, there was no significant difference in the expression of COX-2, VEGF, D114 and Notch-1 mRNA and protein between the diabetic group and the diabetic + distilled water group (P=0.121). There was a significant difference between the other groups (P0.05). The expression of the normal group was least, and the highest expression in the diabetic group and the diabetic + distilled water group, COX-2, VEGF mRNA and protein table. The celecoxib group was higher than the normal group, but lower than the diabetic group and the diabetes + distilled water group, and the highest expression of D114/Notch-1 mRNA and protein in the celecoxib group.
[conclusion]D114 and Notch-1 have played an important role in the formation of retinal neovascularization in diabetic rats. Selective COX-2 Inhibitor Celecoxib can inhibit the expression of VEGF in retina of diabetic rats by activating the D114/ Notch-1 expression.
【學位授予單位】：武漢大學
【學位級別】：博士
【學位授予年份】：2011
【分類號】：R774.1

【共引文獻】

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2 盧百陽;武志峰;;糖尿病視網(wǎng)膜病變發(fā)病機制研究進展[J];國際眼科雜志;2008年11期

3 張莉;蔣升;克麥兒·艾則;;糖尿病大鼠視網(wǎng)膜組織中iNOS的表達及細胞凋亡的研究[J];國際眼科雜志;2009年01期

4 匡洪宇;劉余;郝明;傅錚;馬麗麗;;Exenatide對高糖培養(yǎng)的視網(wǎng)膜神經(jīng)節(jié)細胞的保護作用[J];國際眼科雜志;2011年09期

5 張富花;莊劍波;;環(huán)氧化酶-2與胃癌的研究進展[J];國際內(nèi)科學雜志;2009年07期

6 王嬋娟;艾明;賀濤;邢怡橋;;塞來昔布對糖尿病大鼠視網(wǎng)膜組織生長相關(guān)基因的影響[J];武漢大學學報(醫(yī)學版);2010年06期

7 Joseph I.Helman;;Lymphangiogenesis, Lymphatic Endothelial Cells and Lymphatic Metastasis in Head and Neck Cancer—A Review of Mechanisms[J];International Journal of Oral Science;2010年01期

8 李君靈;曹慶玲;孟紫強;;環(huán)氧化酶-2對細胞動力學和癌變影響的研究進展[J];環(huán)境與職業(yè)醫(yī)學;2008年02期

9 周喜貴;關(guān)向紅;胡杰亮;達明緒;;胃癌組織環(huán)氧化酶-2和血管內(nèi)皮生長因子-C表達的意義[J];蘭州大學學報(醫(yī)學版);2009年03期

10 康艷偉;梅妍;;神經(jīng)遞質(zhì)在糖尿病視網(wǎng)膜病變中作用機制的研究進展[J];四川解剖學雜志;2011年01期

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1 牛玉明;COX-2基因單核苷酸多態(tài)性與頭頸部鱗狀細胞癌易感性的相關(guān)性研究[D];南京醫(yī)科大學;2011年

2 張雪梅;環(huán)氧化酶-2功能性遺傳變異與食管癌及胃癌易感性相關(guān)[D];中國協(xié)和醫(yī)科大學;2006年

3 馬丹;反義環(huán)氧合酶-2基因?qū)θ宋赴┘毎闟GC-7901侵襲力的影響[D];第三軍醫(yī)大學;2007年

4 李小軍;PPARγ、VEGF、VEGFC在人胃癌的表達以及羅格列酮對胃癌細胞生長、新生血管形成和淋巴管形成影響及其機制[D];四川大學;2007年

5 李光源;糖尿病發(fā)展過程中ICAM-1標記物在不同組織血管炎癥反應中的特異性研究[D];吉林大學;2008年

6 黃純海;EGFL7基因在人腦膠質(zhì)瘤中的表達、作用及機制研究[D];中南大學;2008年

7 吳德生;重組pEgr-Endostatin-EGDP質(zhì)粒的輻射誘導特性及其在基因—放射治療中的應用[D];汕頭大學;2007年

8 王俊;HuR介導IL-1β增加NSCLC細胞VEGF-C mRNA穩(wěn)定性的分子機制[D];第三軍醫(yī)大學;2008年

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