本文關(guān)鍵詞： 缺氧 基質(zhì)金屬蛋白酶 間充質(zhì)干細(xì)胞 脈絡(luò)膜新生血管　出處：《眼科新進(jìn)展》2016年05期 　論文類型：期刊論文

【摘要】：目的觀察物理和化學(xué)缺氧模型中小鼠骨髓間充質(zhì)干細(xì)胞(bone marrow-derived mesenchymal stem cells,BMSCs)中基質(zhì)金屬蛋白酶13(matrix metalloproteinase-13,MMP-13)的表達(dá)及其對(duì)猴脈絡(luò)膜-視網(wǎng)膜內(nèi)皮細(xì)胞RF/6A管腔形成能力的影響。方法取C57 BL/6J小鼠骨髓,培養(yǎng)鑒定BMSCs。采用三氣培養(yǎng)箱模擬物理缺氧及氯化鈷(Co Cl2)誘導(dǎo)化學(xué)缺氧。物理缺氧6h、12 h、24 h和48 h后檢測(cè)MMP-13表達(dá)。選表達(dá)最高的處理時(shí)間,檢測(cè)不同濃度Co Cl2(0μmol·L~(-1)、100μmol·L~(-1)、200μmol·L~(-1)、300μmol·L~(-1)及400μmol·L~(-1))處理后MMP-13表達(dá)。檢測(cè)缺氧后BMSCs中缺氧誘導(dǎo)因子-1α(hypoxia-inducible factor-1α,HIF-1α)表達(dá)及BMSCs增殖能力,觀察其條件培養(yǎng)基誘導(dǎo)的RF/6A管腔形成情況。結(jié)果細(xì)胞經(jīng)鑒定符合BMSCs特征。物理缺氧24 h后,MMP-13蛋白表達(dá)量達(dá)高峰(3.16±0.24),較常氧組(1.00±0.12)增加3倍(P0.01)。100μmol·L~(-1)和200μmol·L~(-1)CoC l2組(1.60±0.09、1.64±0.24)中MMP-13表達(dá)較常氧組(1.00±0.20)均增加(均為P0.05),而300μmol·L~(-1)和400μmol·L~(-1)CoC l2組中MMP-13表達(dá)與常氧組相同或稍低。三氣培養(yǎng)箱構(gòu)建的和CoC l2誘導(dǎo)的缺氧環(huán)境下培養(yǎng)24 h后,BMSCs中HIF-1α蛋白表達(dá)較常氧組(1.00±0.23)明顯增加,相對(duì)表達(dá)量分別為3.40±0.26和3.12±0.13(均為P0.01),而兩種缺氧模型間無(wú)明顯差異。在培養(yǎng)24 h時(shí),物理缺氧組(1.53±0.04)和化學(xué)缺氧組(1.31±0.14)均較常氧組(1.04±0.10)細(xì)胞增殖能力增強(qiáng)(均為P0.05),且物理缺氧促增殖效果更明顯。三氣培養(yǎng)箱模擬的物理缺氧組、CoC l2誘導(dǎo)的化學(xué)缺氧組和常氧組RF/6A管腔形成總長(zhǎng)度分別為(5506±380)μm、(5109±558)μm和(3120±300)μm,三氣培養(yǎng)箱模擬的物理缺氧組和CoC l2誘導(dǎo)的化學(xué)缺氧組較常氧組均明顯增加(均為P0.01),而兩種缺氧模型之間差異則無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論 CoC l2和三氣培養(yǎng)箱均可建立缺氧模型,促進(jìn)BMSCs表達(dá)MMP-13,提高其促血管形成能力,提示缺氧可能調(diào)控BMSCs表達(dá)MMPs進(jìn)而影響新生血管發(fā)生發(fā)展。
[Abstract]:Objective to observe the bone marrow-derived mesenchymal stem cells of mouse bone marrow mesenchymal stem cells in physical and chemical hypoxia models. Matrix metalloproteinase 13 matrix metalloproteinase-13. The expression of MMP-13 and its effect on the formation of RF/6A lumen in choroidal and retinal endothelial cells of monkey. Methods the bone marrow of C57 BL/6J mice was harvested. BMSCs was used to culture and identify BMSCs. Three air incubators were used to simulate physical anoxia and cobalt chloride Co Cl2) to induce chemical hypoxia, and physical hypoxia for 6 h and 12 h. After 24 h and 48 h, the expression of MMP-13 was detected. The highest treatment time was selected to detect Co Cl2(0 渭 mol 路L ~ (-1). 100 渭 mol 路L ~ (1) ~ (1) (~ (2) 渭 mol 路L ~ (-1)). The expression of MMP-13 was detected after treatment with 300 渭 mol 路L ~ (-1) and 400 渭 mol 路L ~ (-1), and the expression of hypoxia inducible factor-1 偽 (HIF-1 偽) in BMSCs after hypoxia was detected. Hypoxia-inducible factor-1 偽. The expression of HIF-1 偽 and the proliferative ability of BMSCs were observed. The formation of RF/6A lumen induced by conditioned medium was observed. Results the cells were identified to accord with the characteristics of BMSCs, and the cells were exposed to physical hypoxia for 24 h. The expression of MMP-13 protein reached the peak of 3.16 鹵0.24). Compared with normal oxygen group (1. 00 鹵0. 12), it was increased by 3 times (P0.01U. 100 渭 mol 路L ~ (-1)) and 200 渭 mol 路L ~ (-1) (P < 0. 01) and #number0# 渭 mol 路L ~ (-1) (P < 0. 01). 1.60 鹵0.09. The expression of MMP-13 in 1.64 鹵0.24 was significantly higher than that in normal oxygen group (P0.05). 300 渭 mol 路L ~ (-1) and 400 渭 mol 路L ~ (-1)). The expression of MMP-13 in the CoC L1 group was the same or slightly lower than that in the normoxic group. The expression of HIF-1 偽 protein in BMSCs was significantly higher than that in normal oxygen group (1.00 鹵0.23). The relative expression levels were 3.40 鹵0.26 and 3.12 鹵0.13 (both P 0.01), but there was no significant difference between the two hypoxia models. The proliferation ability of the cells in the physical hypoxia group (1.53 鹵0.04) and the chemical hypoxia group (1.31 鹵0.14) was higher than that in the normal oxygen group (1.04 鹵0.10) (P 0.05). The effect of physical hypoxia on proliferation was more obvious. The total length of RF/6A lumen formation induced by CoC L1 was 5506 鹵380 渭 m in chemical hypoxia group and normoxic group, respectively. 5109 鹵558 渭 m and 3120 鹵300 渭 m respectively. The physical hypoxia group and the chemical hypoxia group induced by CoC l2 were significantly increased compared with the normal oxygen group (P0.01). However, there was no significant difference between the two hypoxia models (P 0.05). Conclusion both CoC l2 and three air incubators can establish hypoxia models and promote the expression of MMP-13 by BMSCs. It is suggested that hypoxia may regulate the expression of MMPs in BMSCs and affect the development of neovascularization.
【作者單位】： 第四軍醫(yī)大學(xué)西京醫(yī)院眼科、全軍眼科研究所;
【基金】：國(guó)家自然科學(xué)基金資助(編號(hào):81200708;81070748;81200151) 第四軍醫(yī)大學(xué)西京醫(yī)院優(yōu)秀人才助推計(jì)劃(2014-2016) 國(guó)家重點(diǎn)基礎(chǔ)研究發(fā)展計(jì)劃(編號(hào):2011CB510200)~~
【分類號(hào)】：R77
【正文快照】： 骨髓間充質(zhì)干細(xì)胞(bone marrow-derived mesen-骨髓貼壁法取C57BL/6J小鼠骨髓細(xì)胞。采用脫頸chymal stem cells,BMSCs)具有取材方便、自體移植椎方法處死小鼠,體積分?jǐn)?shù)75%酒精浸泡5 min,剔不排斥等優(yōu)點(diǎn),已成為細(xì)胞及基因治療的首選種除肌肉和結(jié)締組織后,取出完整股骨和脛骨。

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