| Retinopathy of premature is more common in premature children eye diseases。Mainly due to the current medical standards and the development of perinatal medicine, asubstantial increase in the survival rate of preterm children, especially small children andlow birth weight for gestational age child survival rates increase, thus the probability ofoccurrence of ROP also showed an upward trend. Mechanism of retinopathy in prematurechildren is proliferative retinopathy. The final result of the proliferation of fibrous tissue ofthe retina is to produce traction retinal tissue, eventually leading to blindness in seriousconsequences, so now it becomes the focus of Ophthalmology and neonatology at the sametime and meanwhile been caused many scholars exploration. But its specific pathogenesisis unknown. Based on the obvious abnormal vascular proliferation of ROP, firstly wedesigned ROP animal model of newborn pups, and then transfect angiopoietin-1gene tomarrow mesenchymal stem cells. After that inject them into the model rats at the right time.So we could know the retinal vascular morphology changes and the expression of Ang1byobserving the retinal morphology, hematoxylin-eosin staining, immunohistochemistry andqRT-PCR approach. The main purpose is to discuss impact and effect in ROPneo-vascularization by MSCs carrying Ang-1gene, and provide experimental basis forfuture cell therapy, so as to provide research support for clinical research and basicinformation. The experiment is divided into two parts. In sustained high oxygenenvironment, both injecting Ang-1+MSCs and MSCs alone, can optimizate the retinalvascular morphology, or promote the expression of Ang-1and has some repair function inthe abnormal blood vessels on the retina.Part I: Construction of an animal model of retinopathy of premature childrenWe chose Kunming mice, male and female does not limit and randomly divided intotwo groups, and then were reared in the air and high oxygen, killed on the17thafter birth,We use the modified gelatin ink to perfuse hearts and retinal vascular morphology wasobserved under air group and hyperoxia group. Meanwhile calculate peripheral retinalangiogenesis rate and see if there are statistically significant by using Image-pro plus6.0 processing software and SPSS software. The morphological changes of retinal cell layersand the number of neo-vascularization that breakthrough internal limiting membranethrough paraffin sections and HE staining. Based on the two points, oxygen-inducedanimal model of ROP is successful.Part II: The effect of the MSCs carrying Ang-1gene in the oxygen-induced animal modelof ROP.The first is the recovery of MSCs, subculture and cryopreservation process. Knowingabout the basic nurturing and preservation techniques in this process is the key to theoperation of the experiment. We construct the Ang-1plasmid by Gateway technology andthen packaged with lentiviral and transfect into mice mesenchymal stem cells (Thisresearch was done by LI Qiu-ping). Finally DsRed fluorescent protein technology andmonoclonal were used to select out transfected cells successfully and preparing for the nextintervention. We made ROP mice successfully by using (65%±5%) continuous oxygensupplying. So we could know the retinal vascular morphology changes and the expressionof Ang1by observing the retinal morphology, hematoxylin-eosin staining,immunohisto-chemistry and qRT-PCR approach. The study found that comparng with theair control group, hyperoxia group of retinal blood vessels shaped disorder, had no bloodfilling area caused by blockage of the lumen; Meanwhile prompted by HE staining,thehyperoxia group cells arranged in chaos, and their neovascular endothelial cells broke intothe inner limiting membrane, and the difference was statistically significant; In terms ofmorphology of retinal or HE staining, Ang-1+MSCs injected group was significantlybetter than MSCs group.This indicates that Ang-1and MSCs has synergy role in regulatingangiogenesis. |
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