本文選題：CXC趨化因子受體4 + 氧誘導(dǎo)視網(wǎng)膜病變　； 參考：《武漢大學(xué)》2012年博士論文

【摘要】：視網(wǎng)膜新生血管性疾病是世界上主要的致盲原因之一。此類疾病包括增殖性糖尿病視網(wǎng)膜病變(PDR)、早產(chǎn)兒視網(wǎng)膜病變(ROP)、視網(wǎng)膜中央靜脈阻塞(CRVO)和年齡相關(guān)性黃斑變性(AMD)等。它們共同特征在于視網(wǎng)膜組織相對(duì)缺氧導(dǎo)致了病理性視網(wǎng)膜新生血管的形成。 視網(wǎng)膜血管包含血管發(fā)生(vasculogenesis)和血管新生(neovascularization/angiogenesis)兩種類型。血管發(fā)生是一個(gè)血管從無(wú)到有的過(guò)程,是由骨髓造血干細(xì)胞(HSCs)分化而來(lái)的內(nèi)皮祖細(xì)胞(EPCs),組成相互連接的原始血管網(wǎng),分化為內(nèi)皮細(xì)胞并增殖而形成的血管。血管新生則多被視為一復(fù)雜的病理過(guò)程,例如視網(wǎng)膜受到缺血、缺氧等刺激后,原有血管基底膜降解,內(nèi)皮細(xì)胞增殖、趨化遷移、形成新生血管管腔。大量研究表明,視網(wǎng)膜在受到缺血、缺氧損傷后能釋放出諸多細(xì)胞因子如缺氧誘導(dǎo)因子-1(HIF-1)和血管內(nèi)皮生長(zhǎng)因子(VEGF)、堿性成纖維細(xì)胞生長(zhǎng)因子(bFGF)、表皮生長(zhǎng)因子(EGF)、血小板源性生長(zhǎng)因子(PDGF)、胰島素樣生長(zhǎng)因子(IGFs)、轉(zhuǎn)化生長(zhǎng)因子書(TGF-β)等。這些細(xì)胞生長(zhǎng)因子促使血管內(nèi)皮細(xì)胞有絲分裂和形成新生血管的管腔。SDF-1是一種趨化因子CXC亞家族成員,具有趨化活性,與其受體CXCR4構(gòu)成了SDF-1/CXCR4軸。SDF-1與CXCR4結(jié)合作用啟動(dòng)下游信號(hào)通路,介導(dǎo)炎癥反應(yīng)、引導(dǎo)造血干細(xì)胞遷移及歸巢、以及HIV感染等重要作用。SDF-1/CXCR4還參與了腫瘤、眼部組織新生血管等形成[1,2]�，F(xiàn)SDF-1可以誘導(dǎo)內(nèi)皮祖細(xì)胞到達(dá)視網(wǎng)膜缺血區(qū)域,參與視網(wǎng)膜新生血管發(fā)生[3]AMD3100(商品名為Plerixafor(?))是一種人工合成的CXCR4特異性拮抗劑,能與CXCR4有效結(jié)合,阻斷SDF-1/CXCR4間信號(hào)轉(zhuǎn)導(dǎo),使CXCR4不發(fā)生激動(dòng),已證明可抑制腫瘤新生血管和抗HIV病毒的效果。 然而,CXCR4在缺氧誘導(dǎo)的視網(wǎng)膜新生血管形成過(guò)程中是否起到作用,其作用機(jī)制如何等我們都還不得而知。為此,本課題建立OIR小鼠模型和培養(yǎng)并建立HUVECs缺氧細(xì)胞模型；使用CXCR4的特異性抑制劑AMD3100阻斷SDF-1/CXCR4的信號(hào)轉(zhuǎn)導(dǎo),觀察抑制CXCR4信號(hào)通路對(duì)OIR模型中視網(wǎng)膜新生血管形成的影響,并觀察其對(duì)HUVEC的增殖和表達(dá)新生血管相關(guān)細(xì)胞因子的影響；探討CXCR4在氧誘導(dǎo)視網(wǎng)膜新生血管生成中的作用及其可能的機(jī)制。 第一部分氧誘導(dǎo)視網(wǎng)膜病變小鼠模型和缺氧人臍靜脈內(nèi)皮細(xì)胞模型的建立 目的：采用Smith[4]介紹方法建立OIR小鼠模型；I型膠酶原消化法培養(yǎng)HUVECs并進(jìn)行傳代、鑒定,建立細(xì)胞缺氧模型。方法：取7日齡(P7)C57BL/6小鼠20只,隨機(jī)分成正常組(正常氧環(huán)境下飼養(yǎng)至P17)和OIR組(P7開始每天置于氧濃度在75%±2%飼養(yǎng)至P12,然后取出置于常氧環(huán)境下飼養(yǎng)至P12)。兩組小鼠在P17麻醉后摘取眼球、固定,每組5只小鼠(10只眼)行視網(wǎng)膜切片HE染色,計(jì)算視網(wǎng)膜切片中突破內(nèi)界膜的血管內(nèi)皮細(xì)胞核數(shù)；余5只小鼠(10只眼)采用FITC-Dextran熒光造影視網(wǎng)膜鋪片,定性分析視網(wǎng)膜新生血管情況。無(wú)菌條件下取健康胎兒臍帶一段,用D-Hank's液洗凈臍靜脈管腔,灌注I型膠原酶37℃消化20min后收集細(xì)胞,加入ECM內(nèi)皮細(xì)胞培養(yǎng)基靜置培養(yǎng)、傳代,鏡下觀察細(xì)胞形態(tài)；vWF免疫熒光染色鑒定,繪制細(xì)胞生長(zhǎng)曲線(MTT法檢測(cè))并計(jì)算不同濃度CoCl2缺氧造模后對(duì)細(xì)胞增殖的影響。結(jié)果：OIR模型的50張眼球切片中49張切片有突破內(nèi)界膜的血管內(nèi)皮細(xì)胞核,許多成簇出現(xiàn),有些形成毛細(xì)血管腔,腔內(nèi)可見紅細(xì)胞,視網(wǎng)膜新生血管陽(yáng)性率98%。正常組小鼠平均每張切片突破視網(wǎng)膜內(nèi)界膜的血管內(nèi)皮細(xì)胞核數(shù)為(0.20±0.447)個(gè),OIR模型組平均每張切片的血管內(nèi)皮細(xì)胞核數(shù)為(31.60±2.07)個(gè),差異具有統(tǒng)計(jì)意義(t=-33.09,P0.01)熒光鋪片顯示視網(wǎng)膜大血管不規(guī)則擴(kuò)張,走行迂曲,后極部大片無(wú)灌注區(qū),周邊部可見新生血管叢,伴熒光滲漏。原代培養(yǎng)的細(xì)胞為單層生長(zhǎng),初為圓形,后逐漸伸展呈長(zhǎng)梭形,胞漿豐富,胞核清晰可見,細(xì)胞間有相互連接。vWF抗體免疫熒光染色鑒定示培養(yǎng)細(xì)胞的胞漿內(nèi)含有大量黃綠色熒光顆粒,胞核內(nèi)無(wú)熒光染色。所獲內(nèi)皮細(xì)胞消化、傳代穩(wěn)定,第2-3d生長(zhǎng)速度加快,進(jìn)入對(duì)數(shù)生長(zhǎng)期；CoCl2缺氧造模(0μg/L,20μg/L,50μg/L,100μg/L,200μg/L和400μg/L)對(duì)細(xì)胞生存抑制隨劑量增加而加重。結(jié)論：依照Smith方法可成功建立小鼠OIR模型,視網(wǎng)膜新生血管顯著；酶原消化臍靜脈法可穩(wěn)定獲得HUVECs來(lái)源,給予適量CoCl2模擬化學(xué)缺氧。 第二部分CXCR4受體在氧誘導(dǎo)視網(wǎng)膜病變發(fā)病中的作用研究 目的：探討CXCR4在氧誘導(dǎo)視網(wǎng)膜病變發(fā)生發(fā)展的作用及可能的機(jī)制。方法：100只P7C57BL/6小鼠,隨機(jī)分為5組(n=20)：正常組,OIR模型組,AMD3100大、小劑量玻璃體腔注射組、模型對(duì)照組。除正常組外均建立OIR模型,后三組小鼠P12離開氧箱當(dāng)天給予一次性玻璃體腔注射：AMD3100大劑量(100μg/μL)1μL和AMD3100小劑量(50μg/μL)1μL、模型對(duì)照組(無(wú)菌BSS)1μL。所有小鼠P17戊巴比妥麻醉,每組隨機(jī)抽取5只用于視網(wǎng)膜石蠟切片HE染色和5只用于視網(wǎng)膜FITC-Dextran熒光造影鋪片,計(jì)算突破視網(wǎng)膜內(nèi)界膜內(nèi)皮細(xì)胞數(shù)目并觀察視網(wǎng)膜血管形態(tài)。抽取每組4只小鼠提取視網(wǎng)膜總RNA, RT-PCR半定量分析HIF-1α和VEGF mRNA表達(dá)；剩余每組6只依照western bolt方法檢測(cè)視網(wǎng)膜HIF-1α和VEGF蛋白水平。結(jié)果：正常組(NOR)平均每張切片突破視網(wǎng)膜內(nèi)界膜的內(nèi)皮細(xì)胞核數(shù)為(0.01+/-0.12)個(gè)；缺氧小鼠模型(OIR)平均每張切片突破內(nèi)界膜的內(nèi)皮細(xì)胞核數(shù)為(30.33±1.51)個(gè),與正常組有顯著差異(t=-49.35,P0.01)；AMD3100大劑量(100μg/μL)治療組平均每張切片突破內(nèi)界膜的內(nèi)皮細(xì)胞核數(shù)為(13.50±1.87)個(gè),與正常組差異顯著(t=-17.66,P0.01)；與OIR模型組有顯著差異(t=17.17,P0.01)；AMD3100小劑量(50μg/μL)治療組平均每張切片突破內(nèi)界膜的內(nèi)皮細(xì)胞核數(shù)為(20.83±1.72)個(gè),與正常組有顯著差異,(t=-29.68,P0.01),且與OIR模型組有顯著差異,(t=-10.17,P0.01);模型對(duì)照組平均每張切片突破內(nèi)界膜的內(nèi)皮細(xì)胞核數(shù)為(27.33±2.95)個(gè),與正常組有顯著差異(t=-22.83,P0.01),與OIR模型組無(wú)顯著差異(t=2.10,P0.05)。視網(wǎng)膜FITC熒光鋪片顯示正常組小鼠視網(wǎng)膜結(jié)構(gòu)正常,未見顯著新生血管；OIR模型組視網(wǎng)膜后極部存在大量無(wú)灌注區(qū),大血管迂曲擴(kuò)張、分支減少,網(wǎng)膜中周部有較多新生血管；AMD3100玻璃體腔大劑量注射組視網(wǎng)膜后極部無(wú)灌注區(qū)較OIR模型組的減少,周邊部未見明顯新生血管形成；AMD3100小劑量組的無(wú)灌注區(qū)較OIR模型組減少,新生血管不顯；模型對(duì)照組后極部存在大量無(wú)灌注區(qū),大血管迂曲擴(kuò)張、分支減少,新生血管有熒光滲漏出現(xiàn)。玻璃體腔注射100μg/μL和50μg/μLAMD3100阻斷CXCR4受體能抑制缺氧誘導(dǎo)的HIF-1α和VEGF mRNA和蛋白水平的上調(diào),且100μg/mL AMD3100的抑制作用更強(qiáng),與模型組表達(dá)的差異具有統(tǒng)計(jì)學(xué)意義。結(jié)論：玻璃體腔內(nèi)注射AMD3100可以抑制OIR模型小鼠視網(wǎng)膜新生血管的形成,且可以使缺氧誘導(dǎo)的視網(wǎng)膜組織中的HIF-1α和VEGF mRNA和蛋白表達(dá)量下降,提示SDF-1/CXCR4信號(hào)在缺氧誘導(dǎo)的視網(wǎng)膜新生血管生成中的作用與視網(wǎng)膜組織中HIF-1α和VEGF蛋白表達(dá)的變化有關(guān)。 第三部分CXCR4對(duì)缺氧條件下HUVECs的作用及其機(jī)制 目的：探討SDF-1/CXCR4信號(hào)對(duì)缺氧條件下培養(yǎng)的]HUVECs性狀的改變及其可能機(jī)制。方法：MTT法檢測(cè)AMD3100(50μmol/L,100μmol/L,200μmol/L,400μmol/L,800μmol/L), AMD3100(50μmol/L或100μmol/L)聯(lián)合CoCl2(50μg/L或100μg/L)對(duì)HUVECs生存增殖率的影響；RT-PCR方法檢測(cè)100μg/L CoCl2處理的HUVECs第0h,1h,2h,4h,6h,12h,24h HIF-1α,VEGF mRNA表達(dá)水平；western bolt方法檢測(cè)100μg/L CoCl2處理HUVECs第0h,6h,12h和24h時(shí)HIF-1α和VEGF蛋白表達(dá)；AMD3100(50nM,100nM)預(yù)處理HUVECs1h后給予100μg/L CoCl2,在第24h檢測(cè)HIF-1α和VEGF表達(dá)。結(jié)果：CXCR4受體信號(hào)對(duì)細(xì)胞的增殖存在影響,過(guò)量/AMD3100(800μmol/L)使HUVECs生存率降至正常細(xì)胞的80%。添加/AMD3100(50μmol/L或100μmol/L)加重了CoCl2(50μg/L或100μg/L)對(duì)細(xì)胞生成的抑制。在100μg/L CoCl2處理的0h-24h時(shí)間范圍內(nèi),HIF-1α,VEGF mRNA在第6h-12h出現(xiàn)表達(dá)峰值,在24h表達(dá)水平下調(diào)；HIF-la, VEGF蛋白表達(dá)量在0h-24h間不斷增加；AMD3100(50nM和100nM)預(yù)處理1h后100μg/L CoCl2誘導(dǎo)上調(diào)的HIF-1α和VEGF mRNA和蛋白出現(xiàn)下降,且100nM顯示出更強(qiáng)的抑制作用,與缺氧模型組水平有顯著差異性(P0.01)。結(jié)論：阻斷細(xì)胞的CXCR4信號(hào)通路對(duì)缺氧HUVECs新生血管性改變具有抑制作用。 全文總結(jié) 阻斷SDF-1/CXCR4信號(hào)通路有效抑制了模型小鼠缺氧誘導(dǎo)視網(wǎng)膜新生血管的發(fā)生,缺氧下HUVECs細(xì)胞的增殖以及HIF-lα、VEGF上調(diào)。缺氧環(huán)境使機(jī)體內(nèi)皮細(xì)胞CXCR4受體敏感性上調(diào),其下游信號(hào)活化,促新生血管細(xì)胞因子釋放增加,影響視網(wǎng)膜微環(huán)境的變化,導(dǎo)致視網(wǎng)膜新生血管。
[Abstract]:Retinal neovascular diseases are one of the major causes of blindness in the world. Such diseases include proliferative diabetic retinopathy (PDR), retinopathy of prematurity (ROP), central retinal vein occlusion (CRVO), and age-related macular degeneration (AMD). Their common characteristics are that retinal tissue is histopathologically caused by relative hypoxia. The formation of the neovascularization of the retina.
The retinal vessels include two types of angiogenesis (vasculogenesis) and angiogenesis (neovascularization/angiogenesis). Angiogenesis is a process of vascular endothelial progenitor cells (EPCs) differentiated from bone marrow hematopoietic stem cells (HSCs), forming a interconnected primitive vascular network, differentiating into endothelial cells and proliferating. Angiogenesis is often seen as a complex pathological process, such as the retinal degeneration, endothelial cell proliferation, chemotaxis migration and formation of neovascular lumen after ischemia, hypoxia and other stimuli. A large number of studies show that the retina can release many cytokines such as hypoxia after injury to blood and hypoxia. Inducible factors -1 (HIF-1) and vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), platelet derived growth factor (PDGF), insulin like growth factor (IGFs), transforming growth factor book (TGF- beta), etc.. These cell growth factors induce mitosis and formation of neovascularization in vascular endothelial cells The lumen.SDF-1 is a member of the chemokine CXC subfamily, which has chemotaxis, and its receptor CXCR4 constitutes a SDF-1/CXCR4 axis.SDF-1 and CXCR4 binding to start downstream signaling pathways, mediate inflammatory reactions, guide the migration and homing of hematopoietic stem cells, and HIV infection and other important roles.SDF-1/CXCR4 also participates in the tumor, and the eye tissue is new. [1,2]. SDF-1 can induce endothelial progenitor cells to reach the retinal ischemia area, and participate in retinal neovascularization [3]AMD3100 (commodity name Plerixafor (?)) is a synthetic CXCR4 specific antagonist. It can effectively combine with CXCR4, block the signal transduction between SDF-1/CXCR4 and make CXCR4 not exciting. It has been proved to be possible. Inhibition of neovascularization and anti HIV virus effect.
However, it is not known whether CXCR4 plays a role in the process of hypoxia induced retinal neovascularization, and the mechanism of its action is not known. For this reason, the OIR mouse model was established and the HUVECs hypoxic cell model was developed and the CXCR4 specific inhibitor AMD3100 was used to block the signal transduction of SDF-1/CXCR4 and to observe the inhibition. The effect of CXCR4 signaling pathway on the formation of retinal neovascularization in the OIR model and its effect on the proliferation of HUVEC and the expression of cytokines related to neovascularization, and the role of CXCR4 in the formation of retinal neovascularization in oxygen induced by oxygen and its possible mechanism are also discussed.
The first part is the establishment of oxygen induced retinopathy model and hypoxia human umbilical vein endothelial cell model.
Objective: to establish a OIR mouse model by Smith[4] method, and to cultivate HUVECs with I type enzyme digestion method and carry out the passage, identify and establish the cell hypoxia model. Methods: 20 mice of 7 day old (P7) C57BL/6 mice were randomly divided into normal group (normal oxygen environment, P17) and OIR group (P7 starting at 75% + 2% to P12, at the beginning of P7. Then, the two groups of mice were taken to P12). The two groups of mice were taken eyeball after P17 anesthesia, fixed and 5 mice in each group (10 eyes) were stained with retina slices. The number of vascular endothelial nuclei breaking through the inner boundary membrane in the retinal section was calculated. The remaining 5 mice (10 eyes) were analyzed by FITC-Dextran fluorescein retina paving, qualitative analysis. The condition of retinal neovascularization. Take a healthy fetal umbilical cord under aseptic condition, wash the umbilical vein lumen with D-Hank's liquid, collect I collagenase at 37 C to digest 20min, collect cells, add ECM endothelial cell culture medium, observe cell morphology under microscope, vWF immunofluorescence staining identification, draw cell growth curve (MTT method test). Results: the effects of different concentrations of CoCl2 on cell proliferation were calculated. Results: 49 slices of 50 slice of the eyeball of the OIR model had the vascular endothelial nuclei breaking through the inner boundary membrane, many clusters appeared, some formed capillary cavity, red cells were visible in the cavity, and the positive rate of retinal neovascularization in the normal group of 98%. mice was each cut. The number of vascular endothelial nuclei of the inner boundary membrane of the retina was (0.20 + 0.447). The average number of vascular endothelial nuclei of each slice in OIR model group was (31.60 + 2.07). The difference was statistically significant (t=-33.09, P0.01), which showed irregular expansion of the large retinal vessels in the retina. The primary cultured cells were single layer, and the primary cells were round, and the cells were long shuttle, rich in cytoplasm and clear in the nucleus. There were interconnected.VWF antibody immunofluorescence staining to identify a large number of yellow green fluorescent particles in the cytoplasm of the cultured cells and no fluorescent staining in the nucleus. The cells were digested, the passages were stable, the growth rate of 2-3D accelerated and entered the logarithmic growth period; the CoCl2 hypoxia model (0 mu g/L, 20 mu g/L, 50 mu g/L, 100 mu g/L, 200 mu g/L and 400 mu g/L) aggravated the cell survival inhibition with the dose increase. Conclusion: the mouse OIR model can be established by the Smith method, the retinal neovascularization is significant; the zymogen digestion navel static Pulse method can stabilize the source of HUVECs and give appropriate CoCl2 to simulate chemical hypoxia.
The second part is the role of CXCR4 receptor in the pathogenesis of oxygen induced retinopathy.
Objective: To explore the role and possible mechanism of CXCR4 in the development of oxygen induced retinopathy. Methods: 100 P7C57BL/6 mice were randomly divided into 5 groups (n=20): normal group, OIR model group, AMD3100 large, low dose intravitreal injection group, model control group. The OIR model was established except the normal group, and the later three groups of mice P12 left oxygen box was given the same day Injection of one time vitreous cavity: AMD3100 large dose (100 mu g/ L) 1 L and AMD3100 small dose (50 mu g/ mu L) 1 mu L, model control group (aseptic BSS) 1 u L. all mice P17 pentobarbital anaesthesia, each group randomly selected 5 for retinal paraffin section HE staining and 5 use of retina fluorescein radiography, to calculate the breakthrough retina The number of endothelial cells in the membrane and the retinal vascular morphology were observed. 4 mice in each group were extracted to extract the total retinal RNA, RT-PCR semi quantitative analysis of HIF-1 A and VEGF mRNA expression. The remaining 6 rats were detected the retinal HIF-1 alpha and VEGF protein levels according to the Western bolt method. Results: the average section of the normal group (NOR) broke through the inner boundary membrane of the retina. The number of endothelial nuclei was (0.01+/-0.12), and the average number of endothelial nuclei in the average segment of the hypoxic mouse model (OIR) was (30.33 + 1.51), which was significantly different from the normal group (t=-49.35, P0.01). The number of endothelial nuclei of each section of the AMD3100 large dose (100 mu g/ mu L) group was (13.50 + 1.87), and the number of endothelial nuclei was (13.50 + 1.87) per slice. The difference of the normal group was significant (t=-17.66, P0.01), and there was significant difference with the OIR model group (t=17.17, P0.01), and the number of endothelial nuclei of the average AMD3100 small dose (50 mu g/ mu L) in the treatment group was (20.83 + 1.72), which was significantly different from the normal group, (t= -29.68, P0.01), and there was a significant difference from the OIR model group. In the control group, the average number of endothelial nuclei of the inner boundary membrane was (27.33 + 2.95), which was significantly different from the normal group (t=-22.83, P0.01). There was no significant difference from the OIR model group (t=2.10, P0.05). Retina FITC fluorescent display showed that the normal group of retina was normal in the normal group and no significant neovascularization was found in the normal group, and the posterior retinal pole in the OIR model group was in the posterior pole of the retina. There was a large number of non perfusion areas in the Department. The large blood vessels were tortuous and dilated, the branches were reduced. There were more neovascularization in the periphery of the omentum. The non perfusion area of the posterior retinal pole in the large dose injection group of AMD3100 vitreous cavity was less than that in the OIR model group, and there was no obvious neovascularization in the peripheral part, and the non perfusion area of the small dose group of AMD3100 was less than that of the OIR model group. There was a large number of non perfusion areas in the posterior polar part of the model control group. The large vessels were circuitous and dilated, the branches decreased, and the new blood vessels had fluorescence leakage. The blocking of CXCR4 receptors by intravitreal injection of 100 g/ mu L and 50 u g/ mu LAMD3100 could inhibit the up-regulation of HIF-1 alpha and VEGF mRNA and protein levels induced by hypoxia, and the inhibition of 100 u g/mL AMD3100 The difference between the model group and the model group has statistical significance. Conclusion: Intravitreal injection of AMD3100 can inhibit the formation of retinal neovascularization in OIR model mice, and can reduce the expression of HIF-1 alpha and VEGF mRNA and protein in the retinal tissue induced by hypoxia, and suggest the new SDF-1/CXCR4 signal in the hypoxia induced retina. The role of angiogenesis is related to the expression of HIF-1 alpha and VEGF protein in retina.
The third part is the effect and mechanism of CXCR4 on HUVECs under anoxia condition.
Objective: To investigate the changes and possible mechanisms of the]HUVECs character of SDF-1/CXCR4 signals under hypoxia conditions. Methods: MTT method was used to detect the effect of AMD3100 (50 mu mol/L, 100 mu mol/L, 200 mu mol/L, 400 mu mol/L, 800 micron), AMD3100 (50 mu mol/L or 100 micron) on the survival and proliferation rate. HUVECs 0h, 1H, 2h, 4h, 6h, 12h, 24h HIF-1 alpha were measured by 100 g/L CoCl2. The 4 receptor signal has an effect on the proliferation of cells. Excessive /AMD3100 (800 mu mol/L) reduces the HUVECs survival rate to the normal cell 80%. adding /AMD3100 (50 mu mol/L or 100 mu mol/L) to increase the inhibition of CoCl2 (50 u g/L or 100 mu g/L) to cell formation. Peak value decreased at 24h expression level; HIF-la, VEGF protein expression increased continuously in 0h-24h; AMD3100 (50nM and 100nM) pretreated 1H 100 u g/L CoCl2 induced HIF-1 alpha and VEGF proteins and proteins decreased, and showed a stronger inhibitory effect, and there was a significant difference with the level of the hypoxia model group. Conclusion: blocking. The CXCR4 signaling pathway can inhibit the neovascularization of hypoxic HUVECs.
A summary of the full text
Blocking the SDF-1/CXCR4 signal pathway effectively inhibits the occurrence of retinal neovascularization in the model mice, the proliferation of HUVECs cells under hypoxia and the up regulation of HIF-l alpha and VEGF. The hypoxia environment makes the CXCR4 receptor sensitivity up up, the downstream signal activation, the increase of the release of the neovascularization factor, and the influence of the retinal microring. Changes in the territory of the retina lead to the retinal neovascularization.

【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】：2012
【分類號(hào)】：R774.1

【參考文獻(xiàn)】

相關(guān)期刊論文 前4條

1 徐國(guó)興;陳瑾;鄭學(xué)棟;郭健;謝茂松;;基質(zhì)細(xì)胞衍生因子-1α對(duì)體外培養(yǎng)人視網(wǎng)膜色素上皮細(xì)胞增殖的影響研究[J];國(guó)際眼科雜志;2008年10期

2 陳凌燕;呂林;李永浩;黃新華;張靜琳;李石毅;;增殖性糖尿病視網(wǎng)膜病變玻璃體SDF-1和VEGF的含量分析[J];眼科學(xué)報(bào);2008年01期

3 袁源智;袁非;黎蕾;汪洋;童蓓燕;;基質(zhì)細(xì)胞衍生因子-1在Wistar大鼠視網(wǎng)膜上的生理性表達(dá)[J];眼科研究;2007年07期

4 ;Inhibition of CXCR4 activity with AMD3100 decreases invasion of human colorectal cancer cells in vitro[J];World Journal of Gastroenterology;2008年15期

，

本文編號(hào)：1893232