發(fā)布時(shí)間：2018-05-30 22:14

  本文選題：Basigin + 小膠質(zhì)細(xì)胞��； 參考：《第四軍醫(yī)大學(xué)》2017年博士論文

【摘要】：【研究背景】早產(chǎn)兒視網(wǎng)膜病變(retinopathy of premature,ROP)是一種多發(fā)于早產(chǎn)兒和低體重嬰兒的視網(wǎng)膜血管病變,是目前世界范圍內(nèi)的首要兒童致盲病因。高濃度氧療阻斷了早產(chǎn)兒尚未成熟的視網(wǎng)膜血管的正常發(fā)育進(jìn)程,造成周邊視網(wǎng)膜缺血缺氧,繼而引發(fā)異常新生血管生成,導(dǎo)致視網(wǎng)膜出血、滲出及增殖。因此,缺氧誘發(fā)的異常血管增生是ROP病理發(fā)生的核心機(jī)制之一。視網(wǎng)膜新生血管的形成是機(jī)體應(yīng)對(duì)缺氧環(huán)境的適應(yīng)性反應(yīng),包括內(nèi)皮細(xì)胞與多種細(xì)胞的交互作用,過(guò)程十分復(fù)雜,機(jī)制遠(yuǎn)未闡明。小膠質(zhì)細(xì)胞是定居中樞神經(jīng)系統(tǒng)(central nervous system,CNS)的組織巨噬細(xì)胞,參與免疫調(diào)節(jié)、組織發(fā)育、自身穩(wěn)定及創(chuàng)傷修復(fù)過(guò)程。近來(lái)大量研究選用發(fā)育期的小鼠視網(wǎng)膜作為研究血管生成的模型,結(jié)果證實(shí),視網(wǎng)膜小膠質(zhì)細(xì)胞是血管生成的早期驅(qū)動(dòng)力量。視網(wǎng)膜小膠質(zhì)細(xì)胞與血管緊密相鄰的解剖空間關(guān)系強(qiáng)烈提示,小膠質(zhì)細(xì)胞在血管生成中的重要作用,但是其內(nèi)在機(jī)制尚不清楚。此外,小膠質(zhì)細(xì)胞具有很強(qiáng)的移動(dòng)性,能感知生理及病理刺激,傳遞微環(huán)境的信號(hào),迅速應(yīng)對(duì)局部的趨化信號(hào),通過(guò)分泌細(xì)胞因子及神經(jīng)營(yíng)養(yǎng)因子作用于周?chē)窠?jīng)及血管成分,完成細(xì)胞間的溝通。因此,小膠質(zhì)細(xì)胞在ROP病變中極有可能傳遞局部的缺氧信號(hào),產(chǎn)生適應(yīng)性反應(yīng),促進(jìn)視網(wǎng)膜新生血管生成。Basigin是一種高度糖基化的跨膜糖蛋白,因具有誘導(dǎo)基質(zhì)金屬蛋白酶(matrix metalloproteinase,MMP)生成的能力被命名為MMP誘導(dǎo)因子,簡(jiǎn)稱(chēng)EMMPRIN,也被稱(chēng)為CD147。Basigin具有多重生物功能,如免疫反應(yīng)、基質(zhì)降解及細(xì)胞移行。該分子尤其在腫瘤局部的缺氧環(huán)境下促進(jìn)新生血管生成,從而加強(qiáng)腫瘤的侵襲和轉(zhuǎn)移。它不僅具備蛋白酶誘導(dǎo)作用,促進(jìn)MMP的生成,也能增加腫瘤及內(nèi)皮細(xì)胞的可溶性血管內(nèi)皮生長(zhǎng)因子(vascular endothelial growth factor,VEGF)及血管內(nèi)皮生長(zhǎng)因子受體(vascular endothelial growth factor,VEGFR)2的分泌。在卵巢癌患者,Basigin可通過(guò)可溶性或者以外泌體形式分泌到微環(huán)境中,促進(jìn)腫瘤血管生成。此外,有研究顯示Basigin在單核-巨噬細(xì)胞的表達(dá)上調(diào),增加其炎性活性。Basigin廣泛參與缺氧條件下新生血管的生成,但在以缺氧為核心環(huán)節(jié)的ROP病變中,Basigin是否參與小膠質(zhì)細(xì)胞與血管內(nèi)皮細(xì)胞之間的交互作用還有待研究�！狙芯磕康摹刻接懶∧z質(zhì)細(xì)胞通過(guò)Basigin-2調(diào)控缺氧相關(guān)的視網(wǎng)膜新生血管生成的作用及其機(jī)制,為拓展ROP等缺血缺氧性視網(wǎng)膜新生血管疾病的治療提供實(shí)驗(yàn)依據(jù)。【研究方法】一、體內(nèi)實(shí)驗(yàn):Basigin參與小膠質(zhì)細(xì)胞促進(jìn)氧誘導(dǎo)視網(wǎng)膜病變(oxygen induced retinopathy,OIR)新生血管形成。(1)動(dòng)物模型:建立小鼠OIR模型。(2)組織病理:制作冰凍切片和視網(wǎng)膜鋪片,采用免疫組化及Basigin、IBA-1免疫雙熒光染色,標(biāo)記Basigin及小膠質(zhì)細(xì)胞,觀察兩者在視網(wǎng)膜的表達(dá)及兩者的共定位。(3)Western blot檢測(cè):測(cè)定OIR模型P12和P17,神經(jīng)視網(wǎng)膜低氧誘導(dǎo)因子(hypoxia inducible factor-α,HIF-1α)和Basigin蛋白表達(dá)的變化。二、體外實(shí)驗(yàn):小膠質(zhì)細(xì)胞表達(dá)Basigin,增強(qiáng)視網(wǎng)膜脈絡(luò)膜微血管內(nèi)皮細(xì)胞血管生成能力。(1)細(xì)胞模型:應(yīng)用小膠質(zhì)細(xì)胞BV2細(xì)胞系,物理缺氧模式;(2)構(gòu)建共培養(yǎng)體系:共培養(yǎng)BV2細(xì)胞及視網(wǎng)膜脈絡(luò)膜血管內(nèi)皮細(xì)胞(RF/6A),或應(yīng)用BV2細(xì)胞條件培養(yǎng)基;(3)Western blot檢測(cè):測(cè)定不同處理?xiàng)l件下,小膠質(zhì)細(xì)胞HIF-1α、Basigin、IGF-1、AKT、P-AKT、ERK及P-ERK等蛋白的表達(dá)變化以及RF/6A細(xì)胞VEGF和VEGFR-2的表達(dá)變化;(4)實(shí)時(shí)定量-PCR(quantitative reverse transcription PCR,qRT-PCR):檢測(cè)BV2細(xì)胞在不同處理?xiàng)l件下表達(dá)的Basigin-1和Basigin-2及IGF-1變化,以及RF/6A細(xì)胞VEGF和VEGFR-2的表達(dá)變化;(5)細(xì)胞遷移實(shí)驗(yàn):利用transwell小室測(cè)定RF/6A的遷移能力;(6)管腔形成實(shí)驗(yàn):Matrigel鋪膠,Image Pro Plus軟件測(cè)量管腔形成的長(zhǎng)度。(7)si RNA轉(zhuǎn)染:采用脂質(zhì)體包裹si RNA沉默小膠質(zhì)細(xì)胞Basigin-2的基因表達(dá);(8)細(xì)胞因子測(cè)定:應(yīng)用細(xì)胞因子芯片,篩選出小膠質(zhì)細(xì)胞缺氧后上調(diào),且在Basigin-2敲減后出現(xiàn)下調(diào)的促進(jìn)血管生成的可溶性功能分子。(9)藥物干預(yù):分別應(yīng)用AKT抑制劑(LY294002)及MEK抑制劑(PD98549)抑制AKT或ERK激活,分析其對(duì)IGF-1生成的作用;外源性給予IGF-1蛋白或IGF-1抗體及IGF受體拮抗劑,阻斷或激活I(lǐng)GF-1信號(hào),觀察對(duì)RF/6A管腔形成的影響。(10)統(tǒng)計(jì)學(xué)分析:應(yīng)用統(tǒng)計(jì)學(xué)軟件SPSS 17.0版進(jìn)行分析。多個(gè)樣本組間均數(shù)的比較采用單因素方差分析,任意組間兩兩比較采用Student-Newmann-Keuls(SNK)檢驗(yàn),P0.05為差異有統(tǒng)計(jì)學(xué)意義�！狙芯拷Y(jié)果】一、體內(nèi)實(shí)驗(yàn):Basigin在OIR模型視網(wǎng)膜新生血管簇周?chē)奂男∧z質(zhì)細(xì)胞內(nèi)高表達(dá)。與正常對(duì)照組相比,OIR模型視網(wǎng)膜冰凍切片及視網(wǎng)膜鋪片從不同角度均顯示,視網(wǎng)膜新生血管簇有明顯的IBA-1陽(yáng)性的小膠質(zhì)細(xì)胞聚集,且小膠質(zhì)細(xì)胞標(biāo)志物IBA-1與Basigin在新生血管區(qū)域有顯著的共表達(dá)。OIR組P17神經(jīng)視網(wǎng)膜Basigin的蛋白水平組較正常對(duì)照組顯著增加(P=0.0012)。二、體外實(shí)驗(yàn):缺氧環(huán)境下,小膠質(zhì)細(xì)胞增加Basigin-2表達(dá),促進(jìn)血管內(nèi)皮細(xì)胞血管生成能力及其分子機(jī)制。1.缺氧增加小膠質(zhì)細(xì)胞HIF-1α及Basigin-2的表達(dá)小膠質(zhì)細(xì)胞表達(dá)的HIF-1α及Basigin蛋白隨著物理缺氧時(shí)間的延長(zhǎng)而增加。Basigin的蛋白表達(dá)在缺氧12h(P=0.0097)及24h組(P=0.0003)均較對(duì)照組顯著增加。qRT-PCR檢測(cè)發(fā)現(xiàn),缺氧后的小膠質(zhì)細(xì)胞僅表達(dá)Basigin-2,且隨缺氧時(shí)間延長(zhǎng)而增加。2.缺氧處理后的小膠質(zhì)細(xì)胞促進(jìn)血管內(nèi)皮細(xì)胞的管腔形成缺氧處理后的小膠質(zhì)細(xì)胞與血管內(nèi)皮細(xì)胞共培養(yǎng)或其條件培養(yǎng)基,均能促進(jìn)血管內(nèi)皮細(xì)胞的管腔形成。接種后6h,共培養(yǎng)組管腔形成的長(zhǎng)度均值比對(duì)照組增加60.80%(P=0.0001);條件培養(yǎng)基組均值比對(duì)照組增加52.40%(P=0.0003)。但共培養(yǎng)組與條件培養(yǎng)基組相比無(wú)明顯差別(P=0.4008),提示對(duì)管腔形成的促進(jìn)作用不依賴(lài)于兩種細(xì)胞的直接接觸。3.脂質(zhì)體包裹si RNA成功沉默Basigin-2使用qRT-PCR及Western blot測(cè)定使用脂質(zhì)體包裹的si RNA敲減Basigin后的干涉效果。si RNA Basigin 317和458兩種序列都能明顯下調(diào)Basigin的核酸及蛋白表達(dá),但使用兩種序列聯(lián)合敲減較單獨(dú)敲減組的效果更佳。4.阻斷小膠質(zhì)細(xì)胞Basigin-2表達(dá)抑制血管內(nèi)皮細(xì)胞血管生成能力干涉小膠質(zhì)細(xì)胞Basigin-2表達(dá)后,共培養(yǎng)條件下血管內(nèi)皮細(xì)胞的遷移及管腔形成功能被抑制。敲減后遷移細(xì)胞數(shù)減少65.93%(P=0.0098);接種后6h,管腔形成長(zhǎng)度減少68.27%(P=0.0001)。5.阻斷小膠質(zhì)細(xì)胞的Basigin-2表達(dá)抑制促血管生成因子的生成細(xì)胞因子芯片測(cè)定敲減小膠質(zhì)細(xì)胞Basigin-2基因后血管生成因子的改變,篩選出IGFBP2,IGF-1,Pro-MMP9,VEGFR-1等4個(gè)因子在缺氧后上調(diào);且在Basigin-2敲減后,出現(xiàn)下調(diào)。其中IGF-1表達(dá)改變最為顯著,在缺氧后上調(diào)至基礎(chǔ)值的3.11倍,敲減后下調(diào)為1.33倍。應(yīng)用qRT-PCR及Western blot檢測(cè),BV2敲減Basigin-2后,IGF-1的轉(zhuǎn)錄水平下降(P=0.0001);蛋白表達(dá)下降(P=0.0082)。6.IGF-1促進(jìn)內(nèi)皮細(xì)胞的管腔形成培養(yǎng)基中分別加入IGF-1中和抗體及外源性IGF-1,結(jié)合對(duì)BV2的Basigin-2敲減,評(píng)價(jià)IGF-1對(duì)管腔形成的影響。接種后6h,模擬敲減聯(lián)合IGF-1中和抗體組管腔形成減少28.83%(P=0.0098);Basigin-2敲減聯(lián)合IGF-1可增加管腔形成60.32%(P=0.0002)。7.Basigin-2調(diào)控IGF-1表達(dá)的通路分析分別使用LY294002和PD98549阻斷AKT或ERK通路,qRT-PCR檢測(cè)BV2細(xì)胞不同處理組IGF-1的m RNA水平。Basigin-2敲減及AKT抑制劑能逆轉(zhuǎn)缺氧后上調(diào)的IGF-1趨勢(shì)。Western blot測(cè)定BV2細(xì)胞不同處理組HIF-1,Basigin,IGF-1,P-AKT,AKT,P-ERK及ERK水平。在蛋白水平,缺氧增加IGF-1表達(dá),AKT抑制劑及Basigin-2敲減可以抑制IGF-1的產(chǎn)生,而ERK抑制劑不能抑制IGF-1的表達(dá)。8.IGF-1增強(qiáng)內(nèi)皮細(xì)胞管腔形成能力的分子機(jī)制使用IGF-1受體中和抗體阻斷內(nèi)皮細(xì)胞的IGF-1信號(hào),RF/6A的管腔形成長(zhǎng)度減少101.33%(P=0.0001)。RF/6A表達(dá)的VEGF核酸和蛋白水平無(wú)明顯改變;但VEGFR-2的轉(zhuǎn)錄下調(diào)(P=0.0091),蛋白表達(dá)減少(P=0.0006)�！窘Y(jié)論】本研究首次報(bào)道了Basigin參與小膠質(zhì)細(xì)胞對(duì)氧誘導(dǎo)的視網(wǎng)膜病變新生血管生成的調(diào)控作用,證實(shí)視網(wǎng)膜新生血管簇有明顯的小膠質(zhì)細(xì)胞聚集,且小膠質(zhì)細(xì)胞Basigin表達(dá)水平顯著增加。體外研究證實(shí)了缺氧后小膠質(zhì)細(xì)胞表達(dá)Basigin-2對(duì)血管生成的正性調(diào)控作用,且該作用不依賴(lài)于細(xì)胞間的直接接觸。繼而以脂質(zhì)體包裹si RNA為研究平臺(tái),重點(diǎn)觀察沉默小膠質(zhì)細(xì)胞Basigin-2表達(dá)后對(duì)于血管生成的影響,提出并初步揭示Basigin-2通過(guò)PI3K/AKT途徑增加了促血管生成因子IGF-1的分泌,IGF-1上調(diào)內(nèi)皮細(xì)胞VEGFR-2的生成,增強(qiáng)血管生成的內(nèi)在分子機(jī)制。研究結(jié)果拓展了對(duì)小膠質(zhì)細(xì)胞的認(rèn)識(shí),并且揭示了小膠質(zhì)細(xì)胞表達(dá)Basigin-2作為遞質(zhì),促進(jìn)缺氧相關(guān)的視網(wǎng)膜新生血管生成的調(diào)控作用及機(jī)制,為拓展缺血缺氧性視網(wǎng)膜新生血管疾病的治療提供了新的實(shí)驗(yàn)依據(jù)。
[Abstract]:[background] retinopathy of retinopathy of premature (ROP) is a kind of retinal vascular disease in premature and low weight infants. It is the leading cause of blindness in children worldwide. High concentration oxygen therapy blocks the normal development of retina vascular development in premature infants, resulting in peripheral optic network. Anoxia, which leads to abnormal neovascularization, causes retinal hemorrhage, exudation and proliferation. Therefore, abnormal vascular proliferation induced by hypoxia is one of the core mechanisms of ROP pathology. The formation of the retinal neovascularization is an adaptive response to the anoxic environment, including the interaction of endothelial cells with a variety of cells. The process is very complex and the mechanism is far from clarified. Microglia is the tissue macrophage that settles the central nervous system (CNS). It participates in the process of immunoregulation, tissue development, self stability and trauma repair. Recently, a large number of mouse optic membrane membranes have been used as a model to study angiogenesis. Membrane microglia is an early driving force for angiogenesis. The close spatial relationship between microglia and blood vessels strongly suggests the important role of microglia in angiogenesis, but the intrinsic mechanism of microglia is unclear. In addition, microglia has strong mobility and can perceive physiological and pathological stimuli. The signal of microenvironment is used to respond quickly to the localized chemotactic signal, and by secreting cytokines and neurotrophic factors to the peripheral nerve and blood vessel components to complete the communication between the cells. Therefore, the microglia may transmit the local anoxic signal in the ROP lesion, produce adaptive response, and promote the formation of.Bas in the retina neovascularization. Igin is a highly glycosylated transmembrane glycoprotein. The ability to induce matrix metalloproteinase (matrix metalloproteinase, MMP) generation is named MMP inducer, called EMMPRIN, also known as CD147.Basigin has multiple biological functions, such as immune response, basic degradation and cell migration. This molecule is especially localized in the tumor. In anoxic environment, it promotes neovascularization and strengthens the invasion and metastasis of tumor. It not only has protease induction, promotes the formation of MMP, but also increases the soluble vascular endothelial growth factor (vascular endothelial growth factor, VEGF) and vascular endothelial growth factor receptor (vascular endothelial g) in tumor and endothelial cells (vascular endothelial g). Rowth factor, VEGFR) 2 secretion. In patients with ovarian cancer, Basigin can be secreted into the microenvironment through soluble or exudate forms to promote tumor angiogenesis. In addition, studies have shown that the expression of Basigin in mononuclear macrophages is up-regulated and its inflammatory activity.Basigin is widely involved in the formation of neovascularization under hypoxia. Whether Basigin participates in the interaction between microglia and vascular endothelial cells in ROP lesions with hypoxia as the core link remains to be studied. [Objective] to explore the role and mechanism of microglia cells through Basigin-2 to regulate the formation of retinal neovascularization related to hypoxia, in order to develop ROP and other hypoxic retinal neovascularization. The treatment of vascular disease provides experimental basis. [method] one, in vivo experiment: Basigin participates in microglia to promote the formation of oxygen induced retinopathy of oxygen induced retinopathy (OIR). (1) animal model: establish a mouse OIR model. (2) tissue disease: making frozen section and retina sheet, immunohistochemistry and Bas Igin, IBA-1 immunofluorescence staining, labeling Basigin and microglia, and observing their expression in the retina and their co localization. (3) Western blot detection: the determination of OIR model P12 and P17, the changes of the nerve retinal hypoxia inducible factor (hypoxia inducible factor- alpha, HIF-1 a) and the expression of protein. Two, in vitro experiment: microglia Cell expression Basigin, enhancing the angiogenesis of retinal choroidal microvascular endothelial cells. (1) cell model: the use of microglia BV2 cell line, physical hypoxia model; (2) co culture system: co culture BV2 cells and retinal choroidal vascular endothelial cells (RF /6A), or the application of BV2 cell conditioned medium; (3) Western blot detection: Test: Test The changes in the expression of HIF-1 alpha, Basigin, IGF-1, AKT, P-AKT, ERK and P-ERK in microglia and the changes in the expression of VEGF and VEGFR-2 in RF/6A cells were determined under different treatment conditions. (4) real time quantitative -PCR (Quantitative) Changes, and changes in the expression of VEGF and VEGFR-2 in RF/6A cells; (5) cell migration experiments: using Transwell chamber to determine the migration ability of RF/6A; (6) lumen formation experiment: Matrigel paving, Image Pro Plus software to measure the length of the cavity formation. (7) Si RNA transfection: liposomes encapsulated the gene expression of Si silenced microglia; 8) cytokine assay: using cell factor chip to screen out the up regulation of microglia after hypoxia and down regulation of soluble functional molecules to promote angiogenesis after Basigin-2 knockout. (9) drug intervention: AKT inhibitor (LY294002) and MEK inhibitor (PD98549) were used to inhibit AKT or ERK activation, and the effect on IGF-1 formation was analyzed. Exogenous administration of IGF-1 protein or IGF-1 antibody and IGF receptor antagonist, blocking or activating IGF-1 signal, and observing the effect of RF/6A on the formation of RF/6A. (10) statistical analysis: the application of statistical software SPSS 17 edition for analysis. The comparison of the average number of multiple sample groups using single factor analysis of variance, 22 of any groups using Student-Newmann-K Euls (SNK) test and P0.05 were statistically significant. [results] 1. In vivo experiments: Basigin was highly expressed in the microglia gathered around the neovascular cluster of the retina of the OIR model. Compared with the normal control group, the OIR model retina frozen section and the retina spread were shown from different angles, and the neovascular clusters of the retina were clustered in different angles. IBA-1 positive microglia aggregated, and microglia marker IBA-1 and Basigin had significant co expression in the neovascular region. The protein level group of P17 nerve retina Basigin in group.OIR increased significantly (P=0.0012). Two. In vitro experiment: microglia increased the expression of Basigin-2 and promoted blood vessels in the hypoxic environment. Endothelial cell angiogenesis and its molecular mechanism.1. hypoxia increases HIF-1 A and Basigin-2 in microglia and the expression of HIF-1 A and Basigin protein in microglia and the increase of.Basigin protein expression in the hypoxia 12h (P=0.0097) and 24h group (P=0.0003) increased significantly compared with the control group. It was found that the microglia after anoxia only expressed Basigin-2, and the microglia after hypoxia treatment increased with the prolonged anoxic time to promote the formation of endothelium cells by co culture of microglia and vascular endothelial cells after anoxic treatment of the endothelium of vascular endothelial cells. After 6h, the mean length of the lumen formation in the co culture group was 60.80% higher than that of the control group (P=0.0001), and the mean value of the conditioned medium was 52.40% (P=0.0003) higher than that of the control group, but the co culture group had no significant difference compared with the conditioned medium group (P=0.4008), suggesting that the promoting effect on the formation of the lumen was not dependent on the direct contact with the.3. liposomes of two cells. Parcel Si RNA successfully silence Basigin-2 using qRT-PCR and Western blot to determine the interference effect after Si RNA encapsulated by liposomes.Si RNA Basigin 317 and 458 sequences can obviously reduce the nucleic acid and protein expression, but the effect of the combination of two sequence knockout is better than that of the separate knockout group. The cell Basigin-2 expression inhibited the angiogenesis of vascular endothelial cells and interfered with the expression of Basigin-2 in microglia. The migration of vascular endothelial cells and the function of the lumen formation were inhibited under co culture conditions. The number of migratory cells decreased by 65.93% (P=0.0098) after subtraction. After 6h, the length of the lumen decreased by 68.27% (P=0.0001).5. to block the microglia The expression of Basigin-2 expression inhibited angiogenesis factor by cell factor chip to determine the changes in angiogenesis factor after knockout of Basigin-2 gene in glial cells. 4 factors, such as IGFBP2, IGF-1, Pro-MMP9, VEGFR-1 and so on, were screened out after hypoxia and decreased after Basigin-2 knockout. The most significant change in IGF-1 expression was in the absence of IGF-1 expression. After qRT-PCR and Western blot detection, the transcription level of IGF-1 decreased (P=0.0001) after BV2 knockout Basigin-2, and protein expression decreased (P=0.0082).6.IGF-1 promoted the endothelium cell culture medium with IGF-1 neutralization antibody and exogenous IGF-1, combined with BV2 Asigin-2 knockout to evaluate the effect of IGF-1 on the formation of the lumen. After 6h, the formation of the lumen in the combined IGF-1 neutralization antibody group was reduced by 28.83% (P=0.0098); Basigin-2 knockout combined with IGF-1 could increase the pathway of the cavity formation of 60.32% (P=0.0002).7.Basigin-2 regulated IGF-1. PCR detection of M RNA level.Basigin-2 knockout in different treatment groups of BV2 cells and AKT inhibitor can reverse the increase of IGF-1 trend after hypoxia. ERK inhibitors do not inhibit the molecular mechanism of IGF-1 expression.8.IGF-1 to enhance endothelium cell formation ability by using IGF-1 receptor neutralizing antibodies to block the IGF-1 signal of endothelial cells, and there is no significant change in VEGF nucleic acid and protein levels of 101.33% (P=0.0001).RF/6A expressed by RF/6A in the length of RF/6A, but the transcription of VEGFR-2 (P=0.0091) and protein expression reduction (P=0.0006). [Conclusion] this study was the first to report the role of Basigin in the regulation of microglia induced angiogenesis in oxygen induced retinopathy. It was confirmed that the retinal neovascularization clusters have obvious microglia aggregation, and the expression level of microglia Basigin was significantly increased. The positive regulation of Basigin-2 on angiogenesis after hypoxia is confirmed, and the effect is not dependent on the direct contact between cells. Then the liposomes are wrapped with Si RNA as the research platform, and the effects of Basigin-2 expression on the angiogenesis of silenced microglia are observed, and Basigin-2 is preliminarily revealed through PI3K. The /AKT pathway increases the secretion of angiogenic factor IGF-1, IGF-1 up-regulates the formation of VEGFR-2 in endothelial cells and enhances the intrinsic molecular mechanism of angiogenesis. The results extend the understanding of microglia and reveal the regulation of the expression of Basigin-2 as a transmitter in the hypoxia related retinal neovascularization. The action and mechanism provide a new experimental basis for expanding the treatment of ischemic and hypoxic retinal neovascularization.
【學(xué)位授予單位】：第四軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R774.1
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本文編號(hào)：1957037