三七总皂苷对新生小鼠氧诱导的视网膜病变miR-130a-3p及相关信号通路的影响

发布时间：2018-06-13 13:10

本文选题：早产儿视网膜病变 + miR-130a-3p　；参考：《湖北民族学院》2017年硕士论文

【摘要】：目的:研究三七总皂苷对氧诱导的幼鼠视网膜病变血管新生的影响,证实三七总皂苷活血化瘀作用对氧诱导的幼鼠视网膜病变有着良好的治疗作用,探讨三七总皂苷对高氧诱导的新生小鼠视网膜病变抑制作用的机制为治疗早产儿视网膜病变(Retinopathy of prematurity,ROP)提供新的治疗药物。方法:采用Smith的建模方法高浓度氧气诱导新生小鼠视网膜发生病变建立早产儿视网膜病变模型,随机将实验动物分为五组:正常对照组(A)氧诱导模型组(Oxygen-inducedretinopathy;OIR)(B);腹腔注射4μL组(C);腹腔注射8μL组(D);玻璃体腔注射1μL组(E)(n=15)B、C、D、E组使用氧诱导的视网膜病变的方法造模,其中C、D组在P7-P12每天进行腹腔注射对应剂量的三七总皂苷。E组在P12时进行玻璃体腔注射三七总皂苷1μL。P17时,处死所有幼鼠,使用心脏荧光灌注荧光染色剂异硫氰酸荧光素(Fluorescein isothiocyanate,FITC)的方法做视网膜铺片观察视网膜血管形态,HE染色法观察突出内界膜的视网膜血管内皮细胞核数,Real-time PCR法检测相关基因的表达差异:mi R-130a-3p和类胰岛素生长因子-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)参与VEGF和EGER信号通路,检测两个信号通路的上游VEGFA和EGFR、中游Prkd3,下游核转录因子(Nuclear Factor Kappa B,NF-κB)的表达变化,通过血管内皮生长因子(Vascular endothelial growth factor,VEGF)免疫组化,观察VEGF的阳性表达。结果:HE染色发现C、D、E组较OIR组突出视网膜内界膜的血管内皮细胞核数减少,且E组减少最为明显,差异具有显著性差异(P0.05)。VEGF免疫组化E组的VEGF表达最少。mi R-130a-3p和IGF-1的表达有显著性差异(P0.05)。VEGF信号通路中NF-κB、Prkd3、VEGFA、E组表达差异都有显著性差异(P0.05);EGFR信号通路中NF-κB、Prkd3、EGFR的表达差异具有显著性差异(P0.05)。结论:三七总皂苷能有效的抑制OIR新生小鼠视网膜血管的的新生,改善血管形态的扭曲,增生、无灌注现象;且能影响OIR幼鼠相关基因的表达;表达下调的mi R-130a-3p在使用三七总皂苷后表达上调,同时上调其靶基因Prkd3的表达量;PKC途径参与的VEGF信号通路可以下调OIR幼鼠视网膜VEGFA的表达,PKC途经参与的EGFR信号通路上调OIR幼鼠视网膜EGFR的表达。
[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.
【学位授予单位】：湖北民族学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R285.5

【参考文献】