1、.,脈絡(luò)膜新生血管 CNV,.,CNV的發(fā)生機(jī)制,RPE-Bruch膜-脈絡(luò)膜毛細(xì)血管復(fù)合體改變對CNV的影響,.,.,C57BL/6J（B6） Laser-induced CNV,腹腔麻醉：0.5%戊巴比妥鈉 散瞳：托品酰胺苯腎上腺素 激光：647.1 nm; 50mm spot size; 0.05 s duration; 360 mW. 1PD；3,6,9,12 oclock positions,.,Fundus fluorescein angiography,3 min after intraperitoneal injection of 0.3 mL of 2%fluorescein

2、 sodium was injected into the intraperitoneal cavity of the mice.,.,細(xì)胞,.,新生血管形成過程,內(nèi)皮細(xì)胞的活化（缺氧） (HIF-1)VEGF NO(血管舒張); 內(nèi)皮細(xì)胞通透性增加，有利于血漿向組織擴(kuò)散 血管外基質(zhì)的降解 內(nèi)皮細(xì)胞的增生、移行 新生血管的成熟,VEGF,bFGF,T-PA,u-PA,TNF-a,IL-1,MMP-1,MMP-3,上調(diào),VEGF,NO cGMP,MAPK,bFGF,整合素avB3等,PDGF,aFGF,Ang及受體Tie-1,Tie-2,.,RPE對CNV的雙重調(diào)節(jié)功能,1.促進(jìn)作用 生成促進(jìn)血

3、管內(nèi)皮細(xì)胞增生和移行的VEGF,FGF,TGF-a,TNF-a等等 誘導(dǎo)體內(nèi)血管內(nèi)皮細(xì)胞形成窗孔結(jié)構(gòu)，影響新生血管的成熟 2.抑制作用 直接接觸抑制血管內(nèi)皮細(xì)胞的移行 分泌抑制血管內(nèi)皮細(xì)胞生長的物質(zhì)：TGF-b,PEDF 釋放凝血酶原激活物抑制因子，使纖維酶原向纖溶酶的轉(zhuǎn)化受阻，細(xì)胞外基質(zhì)不能降解,.,巨噬細(xì)胞,激光誘導(dǎo)CNV，第一天巨噬細(xì)胞出現(xiàn)，第三天達(dá)到高峰，5-7天后開始減少。,VEGF，MMP-9 蛋白水解酶（玻璃膜）,IL-1a,IL-1b,TNF-a IL-8,MCP-1,生成血管活性因子,巨噬細(xì)胞+RPE,.,參與血管內(nèi)皮生長因子誘發(fā)脈絡(luò)膜新生血管的信號通路,MAPK途徑 PI-

4、3K-AKt/PKB途徑 鈣離子依賴性激酶途徑 NO途徑,.,P38內(nèi)皮細(xì)胞移行,VEGF誘導(dǎo)活化:ERK,JNK,.,PI-3K:CEC增生,ILK：內(nèi)皮細(xì)胞增生和移行,.,.,.,轉(zhuǎn)錄因子,STAT3 NF-kappa-B HIF-1,.,STAT3,Signal transducer and transcription activator 退化期CNV的RPE細(xì)胞中，有STAT3的高表達(dá)，而在已纖維化的盤狀瘢痕中，STAT3的表達(dá)呈陰性，提示STAT3的活化與CNV的纖維化有關(guān)。,.,STAT3+CNV,1.A novel protective role for the innate i

5、mmunity Toll-Like Receptor 3 (TLR3) in the retina via Stat3. Mol Cell Neurosci. 2014 Sep 28;63C:38-48 IF:3.73 2.CNTF-mediated protection of photoreceptors requires initial activation of the cytokine receptor gp130 in Mller glial cells. Proc Natl Acad Sci U S A. 2013 Nov 19;110(47):E4520-9 IF:9.81 3.

6、Toll-like receptor 3 (TLR3) protects retinal pigmented epithelium (RPE) cells from oxidative stress through a STAT3-dependent mechanism. Mol Immunol. 2013 Jun;54(2):122-31 IF:3.0 4.Suppression of choroidal neovascularization through inhibition of APE1/Ref-1 redox activity. Invest Ophthalmol Vis Sci.

7、 2014 Jun 26;55(7):4461-9 IF.3.66 5.IL-27 inhibits pathophysiological intraocular neovascularization due to laser burn. J Leukoc Biol. 2012 Feb;91(2):267-73 IF:4.3 6.Interleukin-6 receptor-mediated activation of signal transducer and activator of transcription-3 (STAT3) promotes choroidal neovascula

8、rization. Am J Pathol. 2007 Jun;170(6):2149-58 IF:4.6 7.Hyperglycaemia exacerbates choroidal neovascularisation in mice via the oxidative stress-induced activation of STAT3signalling inRPEcells. PLoS One. 2012;7(10):e47600. doi: 10.1371/journal.pone.0047600. Epub 2012 Oct 19,.,NF-kappa-B,NFKB is a t

9、ranscription regulator that is activated by various intra- and extra-cellular stimuli such as cytokines, oxidant-free radicals, ultraviolet irradiation, and bacterial or viral products. Activated NFKB translocates into the nucleus and stimulates the expression of genes involved in a wide variety of

10、biological functions. Inappropriate activation of NFKB has been associated with a number of inflammatory diseases while persistent inhibition of NFKB leads to inappropriate immune cell development or delayed cell growth. NFB受多種細(xì)胞內(nèi)和細(xì)胞外的刺激如細(xì)胞因子，氧化劑自由基，紫外線照射，細(xì)菌性或病毒性產(chǎn)物激活的轉(zhuǎn)錄調(diào)節(jié)子。激活的NFB易位到細(xì)胞核中，并刺激參與多種生物功能的

11、基因的表達(dá)。NFB的不適當(dāng)活化已經(jīng)與許多炎癥性疾病相關(guān)，而NFKB的持久性抑制導(dǎo)致不適當(dāng)?shù)拿庖呒?xì)胞的發(fā)育或延緩細(xì)胞生長。,.,NF-kappa-B,1.Roles for the ubiquitin-proteasome pathway in protein quality control and signaling in the retina: implications in the pathogenesis of age-related macular degeneration. Mol Aspects Med. 2012 Aug;33(4):446-66 IF:10.3 2.Macula

12、r pigment lutein is antiinflammatory in preventing choroidal neovascularization. Arterioscler Thromb Vasc Biol. 2007 Dec;27(12):2555-62 IF:5.53 3.IKK2 inhibition attenuates laser-induced choroidal neovascularization. PLoS One. 2014;9(1):e87530 IF:3.53 4.TNF- decreases VEGF secretion in highly polari

13、zed RPE cells but increases it in non-polarized RPE cells related to crosstalk between JNK and NF-B pathways. PLoS One. 2013;8(7):e69994 IF:3.53 5.NLRP3 inflammasome activation in retinal pigment epithelial cells by lysosomal destabilization: implications for age-related macular degeneration. Invest

14、 Ophthalmol Vis Sci. 2013 Jan 7;54(1):110-20 IF:3.66 6.Regulatory role of HIF-1alpha in the pathogenesis of age-related macular degeneration (AMD). Ageing Res Rev. 2009 Oct;8(4):349-58 IF:7.63 7.Roles of NFB-miR-29s-MMP-2 circuitry in experimental choroidal neovascularization. J Neuroinflammation. 2

15、014 May 15;11:88 IF:4.9,.,HIF-1,Functions as a master transcriptional regulator of the adaptive response to hypoxia. Under hypoxic conditions, activates the transcription of over 40 genes, including erythropoietin, glucose transporters,glycolytic enzymes, vascular endothelial growth factor, HILPDA,

16、and other genes whose protein products increase oxygen delivery or facilitate metabolic adaptation to hypoxia. Plays an essential role in embryonic vascularization, tumor angiogenesis and pathophysiology of ischemic disease. 缺氧適應(yīng)性反應(yīng)的主轉(zhuǎn)錄調(diào)節(jié)。在缺氧條件下，激活了40個基因，包括促紅細(xì)胞生成素，葡萄糖轉(zhuǎn)運(yùn)，糖酵解酶，血管內(nèi)皮生長因子，乳過氧化物酶，和其他基因，其蛋

17、白產(chǎn)物增加氧氣傳遞或促進(jìn)代謝適應(yīng)低氧的轉(zhuǎn)錄。在胚胎血管生成，腫瘤血管生成和缺血性疾病的病理生理學(xué)中起重要作用。,.,HIF-1,1.Decorin inhibits angiogenic potential of choroid-retinal endothelial cells by downregulating hypoxia-induced Met, Rac1, HIF-1 and VEGF expression in cocultured retinal pigment epithelial cells. Exp Eye Res. 2013 Nov;116:151-60 IF:3.02

18、 2.Methallothionein-3 contributes to vascular endothelial growth factor induction in a mouse model of choroidal neovascularization. Metallomics. 2013 Oct;5(10):1387-96 IF:3.98 3.Impacts of hypoxia-inducible factor-1 knockout in the retinal pigment epithelium on choroidal neovascularization. Invest Ophthalmol Vis Sci. 2012 Sep 14;53(10):6197-206 IF:3.66 4.Influence of Dll4 via HIF-1-VEGF signaling on the angiogenesis of choroidal neovascularization under hypoxic condi