【摘要】：目的觀察Bevacizumab對(duì)人視網(wǎng)膜色素上皮細(xì)胞纖維化相關(guān)因子基因及蛋白表達(dá)的影響,探討B(tài)evacizumab在新生血管纖維化中的作用。方法用含終濃度0.25 g·L-1Bevacizumab的DMED/F12培養(yǎng)液培養(yǎng)ARPE-19細(xì)胞,按Bevacizumab處理的時(shí)間分為0 h、6 h、12 h、24 h和48 h共5組,0 h為對(duì)照組。不同時(shí)間點(diǎn)收集細(xì)胞及上清。用逆轉(zhuǎn)錄聚合酶鏈反應(yīng)檢測(cè)ARPE-19細(xì)胞中結(jié)締組織生長(zhǎng)因子(connective tissue growth factor,CTGF)、堿性成纖維細(xì)胞生長(zhǎng)因子(basic fibroblast growth factor,bFGF)、轉(zhuǎn)化生長(zhǎng)因子-β2(transforming growth factorβ2,TGF-β2)和基質(zhì)金屬蛋白酶-2(matrix metalloproteinase 2,MMP-2)的mRNA相對(duì)表達(dá)量。采用ELISA檢測(cè)上清中CTGF、bFGF和TGF-β2的蛋白濃度,應(yīng)用免疫蛋白印跡法(Western blotting)檢測(cè)MMP-2蛋白的表達(dá)。結(jié)果 Bevacizumab作用后6 h、12 h、24 h和48 h,CTGF、bFGF、TGF-β2及MMP-2的mRNA相對(duì)表達(dá)量分別為CTGF:1.694±0.160、1.994±0.221、1.733±0.057、1.397±0.075,bFGF:1.720±0.068、2.144±0.080、1.893±0.068、1.372±0.076,TGF-β2:1.356±0.070、1.737±0.515、1.576±0.098、1.439±0.050,MMP-2:1.414±0.115、1.644±0.094、1.435±0.075、1.285±0.024,與對(duì)照組相比均升高,差異均有統(tǒng)計(jì)學(xué)意義(均為P0.05)。ELISA結(jié)果顯示Bevacizumab作用后6 h、12 h、24 h和48 h,CTGF、bFGF、TGF-β2蛋白的表達(dá)量與對(duì)照組相比均升高,差異均有統(tǒng)計(jì)學(xué)意義(均為P0.05);對(duì)照組及各時(shí)間組蛋白濃度分別為CTGF:(31.949±0.388)ng·L-1、(48.697±0.863)ng·L-1、(56.847±0.499)ng·L-1、(51.753±1.395)ng·L-1、(43.085±1.240)ng·L-1,bFGF:(26.660±1.360)ng·L-1、(46.529±3.297)ng·L-1、(55.883±1.705)ng·L-1、(47.937±1.303)ng·L-1、(36.894±2.532)ng·L-1,TGF-β2:(26.982±2.404)ng·L-1、(35.452±1.360)ng·L-1、(46.787±0.896)ng·L-1、(43.683±1.630)ng·L-1、(38.008±1.110)ng·L-1。Western blotting結(jié)果顯示,Bevacizumb處理后6 h、12 h、24 h、48 h MMP-2蛋白含量升高,組間比較差異有統(tǒng)計(jì)學(xué)意義(F=2620.429,P=0.000)。結(jié)論 Bevacizumab能夠上調(diào)人RPE細(xì)胞中CTGF、bFGF、TGF-β2及MMP-2的表達(dá),可能有促纖維化形成的作用。
[Abstract]:Objective to observe the effect of Bevacizumab on the expression of fibrosis related factor gene and protein in human retinal pigment epithelial cells (RPE) and to explore the role of Bevacizumab in neovascularization. Methods ARPE-19 cells were cultured in DMED/F12 medium containing 0.25 g 路L-1Bevacizumab. According to the treatment time of Bevacizumab, ARPE-19 cells were divided into 5 groups for 0 h, 6 h, 12 h, 24 h and 48 h, and 0 h as control group. At different time points, cells and smears were collected. Connective tissue growth factor (connective tissue growth factor,CTGF, basic fibroblast growth factor (basic fibroblast growth factor,bFGF and transforming growth factor-尾 2 (transforming growth factor 尾 2 in ARPE-19 cells were detected by reverse transcription polymerase chain reaction (RT-PCR). The mRNA expression of TGF- 尾 2) and matrix metalloprotease-2 (matrix metalloproteinase 2 (MMP 鈮，

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