本文選題：早產(chǎn)兒視網(wǎng)膜病變 + miR-130a-3p��； 參考：《湖北民族學(xué)院》2017年碩士論文

【摘要】：目的:研究三七總皂苷對氧誘導(dǎo)的幼鼠視網(wǎng)膜病變血管新生的影響,證實三七總皂苷活血化瘀作用對氧誘導(dǎo)的幼鼠視網(wǎng)膜病變有著良好的治療作用,探討三七總皂苷對高氧誘導(dǎo)的新生小鼠視網(wǎng)膜病變抑制作用的機制為治療早產(chǎn)兒視網(wǎng)膜病變(Retinopathy of prematurity,ROP)提供新的治療藥物。方法:采用Smith的建模方法高濃度氧氣誘導(dǎo)新生小鼠視網(wǎng)膜發(fā)生病變建立早產(chǎn)兒視網(wǎng)膜病變模型,隨機將實驗動物分為五組:正常對照組(A)氧誘導(dǎo)模型組(Oxygen-inducedretinopathy;OIR)(B);腹腔注射4μL組(C);腹腔注射8μL組(D);玻璃體腔注射1μL組(E)(n=15)B、C、D、E組使用氧誘導(dǎo)的視網(wǎng)膜病變的方法造模,其中C、D組在P7-P12每天進行腹腔注射對應(yīng)劑量的三七總皂苷。E組在P12時進行玻璃體腔注射三七總皂苷1μL。P17時,處死所有幼鼠,使用心臟熒光灌注熒光染色劑異硫氰酸熒光素(Fluorescein isothiocyanate,FITC)的方法做視網(wǎng)膜鋪片觀察視網(wǎng)膜血管形態(tài),HE染色法觀察突出內(nèi)界膜的視網(wǎng)膜血管內(nèi)皮細胞核數(shù),Real-time PCR法檢測相關(guān)基因的表達差異:mi R-130a-3p和類胰島素生長因子-1(Insulin-like growth factor-1,IGF-1),mi R-130a-3p的靶基因蛋白激酶D3(Protein Kinase D3,Prkd3)參與VEGF和EGER信號通路,檢測兩個信號通路的上游VEGFA和EGFR、中游Prkd3,下游核轉(zhuǎn)錄因子(Nuclear Factor Kappa B,NF-κB)的表達變化,通過血管內(nèi)皮生長因子(Vascular endothelial growth factor,VEGF)免疫組化,觀察VEGF的陽性表達。結(jié)果:HE染色發(fā)現(xiàn)C、D、E組較OIR組突出視網(wǎng)膜內(nèi)界膜的血管內(nèi)皮細胞核數(shù)減少,且E組減少最為明顯,差異具有顯著性差異(P0.05)。VEGF免疫組化E組的VEGF表達最少。mi R-130a-3p和IGF-1的表達有顯著性差異(P0.05)。VEGF信號通路中NF-κB、Prkd3、VEGFA、E組表達差異都有顯著性差異(P0.05);EGFR信號通路中NF-κB、Prkd3、EGFR的表達差異具有顯著性差異(P0.05)。結(jié)論:三七總皂苷能有效的抑制OIR新生小鼠視網(wǎng)膜血管的的新生,改善血管形態(tài)的扭曲,增生、無灌注現(xiàn)象;且能影響OIR幼鼠相關(guān)基因的表達;表達下調(diào)的mi R-130a-3p在使用三七總皂苷后表達上調(diào),同時上調(diào)其靶基因Prkd3的表達量;PKC途徑參與的VEGF信號通路可以下調(diào)OIR幼鼠視網(wǎng)膜VEGFA的表達,PKC途經(jīng)參與的EGFR信號通路上調(diào)OIR幼鼠視網(wǎng)膜EGFR的表達。
[Abstract]:Objective: to study the effect of panax notoginseng saponins on angiogenesis of oxygen-induced retinopathy in young rats. To explore the inhibitory mechanism of panax notoginseng saponins on hyperoxia-induced retinopathy in neonatal mice. Methods: the model of retinopathy of premature infant was established by Smith's modeling method, which was induced by high oxygen concentration in newborn mice. The experimental animals were randomly divided into five groups: Oxygen-induced retinopathy model group (Oxygen-induced retinopathy group); intraperitoneal injection of 4 渭 L group Con; intraperitoneal injection of 8 渭 L group; intravitreal injection of 1 渭 L group (1 渭 L group); oxygen-induced retinopathy in 1 渭 L group. All the young rats were killed by intraperitoneal injection of panax notoginseng saponins (P7-P12) and panax notoginseng saponins (panax notoginseng) 1 渭 L. P17 in group E at P12. Using Fluorescein isothiocyanate FITC as a New fluorescent Perfusion staining method for Retinal Vascular Morphology and HE staining to observe the number of Retinal Endothelial Nucleus in the Endothelial membrane of the protrusion and Real-time PCR Assay for the Detection of the correlation between Retinal Vascular Endothelial Nucleus and Retinal Vascular Endothelial Nucleus The difference in gene expression: mi R-130a-3p and insulin-like growth factor-1 (IGF-1) mi R-130a-3p are involved in the signal pathway of VEGF and EGER. The expression of VEGFA and EGFR in upstream, Prkd3 and nuclear factor Kappa BnNF- 魏 B in the middle reaches of the two signaling pathways were detected. The positive expression of VEGF was observed by immunohistochemical staining of vascular endothelial growth factor (Vascular endothelial growth factor). Results the number of vascular endothelial nuclei in group E was significantly lower than that in group OIR, and the number of endothelial nuclei in group E was significantly lower than that in group E, and the number of vascular endothelial nuclei in group E was significantly lower than that in group OIR. There was a significant difference in the expression of VEGF between P0.05A and VEGF immunohistochemical E group. There was a significant difference in the expression of VEGF between P0.05A and IGF-1. There were significant differences in the expression of NF- 魏 B, Prkd3VEGFAE and the expression of NF- 魏 B, Prkd3EGFR and EGFR in P0.05T / EGFR signaling pathway (P < 0.05). Conclusion: panax notoginseng saponins can effectively inhibit the retinal angiogenesis, improve the morphologic distortion, proliferation and no perfusion, and affect the expression of related genes in young OIR mice. The down-regulated expression of mi R-130a-3p was up-regulated after panax notoginseng saponins were used. At the same time, the expression of Prkd3 and the expression of VEGFA in the retina of young OIR rats were down-regulated by the VEGF signaling pathway involved by PKC pathway. The expression of EGFR in the retina of young OIR rats was up-regulated by PKC pathway.
【學(xué)位授予單位】：湖北民族學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R285.5

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