本文選題：糖尿病性視網(wǎng)膜病變 + 促紅細(xì)胞生成素��； 參考：《武漢大學(xué)》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(guó)威脅人類健康的常見多發(fā)病,與此相應(yīng),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(jī)制還不完全清楚,大量的研究認(rèn)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(zhǎng)因子的作用有關(guān),同時(shí)炎癥及免疫反應(yīng)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(dá) [目的]通過檢測(cè)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(xì)胞生成素(EPO)、生長(zhǎng)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(duì)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(duì)照組。采用酶聯(lián)免疫吸附試驗(yàn)(ELISA)檢測(cè)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(duì)照組(23.52±7.27)mIU/ml。玻璃體EPO含量：PDR組(461.36±101.20)mIU/ml,對(duì)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(duì)照組(55.68±22.53) pg/ml。玻璃體Gro-a含量：PDR組(327.74±216.29) pg/ml,對(duì)照組(81.95±47.59)pg/ml。3組血漿樣本間EPO含量差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05), Gro-a含量差異有統(tǒng)計(jì)學(xué)意義(P0.05)。3組玻璃體樣本內(nèi)PDR組與對(duì)照組2組間EPO、Gro-α差異均有顯著統(tǒng)計(jì)學(xué)意義(P0.05)。各組血漿、玻璃體內(nèi)EPO、Gro-α含量與相應(yīng)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(dá)及塞來(lái)昔布 對(duì)GRO-α、EPO表達(dá)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(dá),以及選擇性環(huán)氧合酶-2 (Cyclooxygenase-2, COX-2)抑制劑(塞來(lái)昔布)對(duì)糖尿病大鼠視網(wǎng)膜組織中GRO-α和EPO表達(dá)的影響。 [方法]將40只SD大鼠隨機(jī)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(lái)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(dǎo)建立糖尿病大鼠動(dòng)物模型,飼養(yǎng)12周后處死大鼠,取視網(wǎng)膜,SYBR熒光實(shí)時(shí)定量PCR檢測(cè)視網(wǎng)膜組織中COX-2、血管內(nèi)皮生長(zhǎng)因子(vascular endothelial growth factor, VEGF)、EPO和GRO-a mRNA的表達(dá)變化,western-blotting檢測(cè)大鼠視網(wǎng)膜中COX-2、VEGF、EPO和GRO-a蛋白的表達(dá)變化。通過Quantity One凝膠圖像分析軟件進(jìn)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(jì)學(xué)意義。 [結(jié)果]12周后,COX-2, VEGF, EPO及GRO-a mRNA和蛋白質(zhì)的表達(dá)在糖尿病組和糖尿病+蒸餾水組最高,糖尿病+塞來(lái)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(wú)統(tǒng)計(jì)學(xué)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(lái)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(dá),從而抑制糖尿病大鼠視網(wǎng)膜組織中VEGF的表達(dá),最終起到抗新生血管形成的作用。 第三部分塞來(lái)昔布對(duì)糖尿病大鼠視網(wǎng)膜組織中d114、Notch-1表達(dá)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(lái)昔布)對(duì)糖尿病大鼠視網(wǎng)膜組織中Delta樣配體4- Notch信號(hào)(delta-like 4 ligan-Notch, D114/Notch-1)通路的影響。 [方法]將40只SD大鼠隨機(jī)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(lái)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(dǎo)建立糖尿病動(dòng)物模型,飼養(yǎng)12周后處死大鼠,取視網(wǎng)膜組織,SYBR熒光實(shí)時(shí)定量PCR檢測(cè)視網(wǎng)膜組織中COX-2、血管內(nèi)皮生長(zhǎng)因子(vascular endothelial growth factor,VEGF)以及D114/Notch-1 mRNA的表達(dá)變化,western-blotting檢測(cè)大鼠視網(wǎng)膜組織中COX-2、VEGF以及D114/Notch-1蛋白的表達(dá)。通過Quantity One凝膠圖像分析軟件進(jìn)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(jì)學(xué)意義。 [結(jié)果]12周后,COX-2、VEGF、D114及Notch-1 mRNA和蛋白質(zhì)的表達(dá)在糖尿病組和糖尿病+蒸餾水組間差異無(wú)統(tǒng)計(jì)學(xué)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(dá)最少,糖尿病組和糖尿病+蒸餾水組表達(dá)最高,COX-2、VEGF mRNA和蛋白質(zhì)表達(dá)在塞來(lái)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(lái)昔布用藥組表達(dá)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(lái)昔布可以通過激活糖尿病大鼠視網(wǎng)膜中的D114/ Notch-1表達(dá),從而抑制糖尿病大鼠視網(wǎng)膜組織中的VEGF表達(dá)。
[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(xué)位授予單位】：武漢大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2011
【分類號(hào)】：R774.1

【共引文獻(xiàn)】

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9 居e，

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