發(fā)布時間：2018-01-26 01:54

  本文關(guān)鍵詞： 康柏西普 血小板衍生生長因子受體抑制劑 缺氧 人視網(wǎng)膜色素上皮細胞 血管內(nèi)皮生長因子　出處：《眼科新進展》2017年12期 　論文類型：期刊論文

【摘要】：目的探討康柏西普聯(lián)合血小板衍生生長因子(platelet derived growth factor,PDGF)受體抑制劑對CoCl_2誘導的人視網(wǎng)膜色素上皮細胞ARPE-19細胞增殖、遷移以及血管內(nèi)皮生長因子(vascular endothelial growth factor,VEGF)m RNA及蛋白表達的影響。方法體外培養(yǎng)ARPE-19細胞,取對數(shù)生長良好的細胞,用不同濃度CoCl_2誘導培養(yǎng),細胞增殖與CCK-8法檢測ARPE-19細胞增殖活性,篩選最佳CoC l2濃度用于構(gòu)建缺氧模型;將培養(yǎng)的ARPE-19細胞分為正常組、缺氧組、不同濃度康柏西普組,CCK-8法檢測不同濃度康柏西普對缺氧損傷后ARPE-19細胞增殖的影響,篩選最佳康柏西普濃度;將培養(yǎng)的ARPE-19細胞分為正常組、缺氧組、不同濃度PDGF受體抑制劑組,CCK-8法檢測不同濃度PDGF受體抑制劑對缺氧損傷后ARPE-19細胞增殖的影響,篩選最佳PDGF受體抑制劑濃度;將培養(yǎng)的ARPE-19細胞分為正常組、缺氧組、20 mg·L~(-1)康柏西普組、20μmol·L~(-1)PDGF受體抑制劑組、康柏西普聯(lián)合PDGF受體抑制劑組(最佳藥物濃度組合);CCK-8法檢測缺氧損傷后各組ARPE-19細胞增殖活性,Transwell小室檢測細胞的遷移情況,ELISA法檢測細胞上清液中VEGF蛋白濃度,應(yīng)用實時熒光定量PCR法檢測細胞VEGF mR NA的表達水平。結(jié)果 CCK-8篩選結(jié)果顯示,100μmol·L~(-1)CoC l2處理組ARPE-19細胞的吸光度值最大(1.063±0.031),設(shè)立為缺氧細胞模型濃度。CCK-8篩選結(jié)果顯示,20 mg·L~(-1)康柏西普組細胞增殖率為(94.58±3.80)%、20μmol·L~(-1)PDGF受體抑制劑組細胞增殖率為(96.72±5.44)%,效果均較其他相應(yīng)組好,選取該濃度進行后續(xù)實驗。析因分析發(fā)現(xiàn)缺氧損傷后各藥物處理組(康柏西普、PDGF受體抑制劑、康柏西普聯(lián)合PDGF受體抑制劑)與缺氧組比較,細胞增殖活性下降,細胞遷移數(shù)降低,VEGF蛋白濃度亦下降,其中康柏西普聯(lián)合PDGF受體抑制劑組下降最明顯,康柏西普組及兩藥聯(lián)合處理組ARPE-19細胞VEGF mR NA表達較缺氧組降低。結(jié)論缺氧損傷后可誘導ARPE-19細胞增殖、遷移,VEGF蛋白相對表達量增加,VEGF mR NA表達上調(diào);康柏西普聯(lián)合PDGF受體抑制劑對缺氧損傷后ARPE-19細胞的增殖、遷移及VEGF蛋白、VEGF mR NA的表達有抑制作用,作用效果優(yōu)于單獨應(yīng)用康柏西普。
[Abstract]:Objective to investigate the combination of Compactsep and platelet derived growth factor. PDGF receptor inhibitor could induce the proliferation of human retinal pigment epithelium (ARPE-19) cells induced by CoCl_2. Migration and vascular endothelial growth factor. Methods ARPE-19 cells were cultured in vitro. The cells with good logarithmic growth were cultured with different concentrations of CoCl_2. Cell proliferation and CCK-8 assay were used to detect the proliferative activity of ARPE-19 cells, and the optimal concentration of CoC l2 was selected to construct anoxic model. The cultured ARPE-19 cells were divided into three groups: normal group, hypoxia group, and different concentrations of Compactopril (CCK-8). The effects of different concentrations of Compactopril on the proliferation of ARPE-19 cells after hypoxia injury were detected by CCK-8 method. Screening of the best concentration of Compactopril; The cultured ARPE-19 cells were divided into normal group hypoxia group and different concentration of PDGF receptor inhibitor group. The effects of different concentrations of PDGF receptor inhibitors on the proliferation of ARPE-19 cells after hypoxia injury were detected by CCK-8 assay, and the best concentration of PDGF receptor inhibitors was selected. The cultured ARPE-19 cells were divided into two groups: normal group, hypoxia group (20 mg 路L ~ (-1)) Compactopril group (n = 20 渭 mol 路L ~ (-1)). Compactopril combined with PDGF receptor inhibitor group (best drug concentration combination); CCK-8 assay was used to detect the proliferation activity of ARPE-19 cells in each group after hypoxia injury. ELISA assay was used to detect the concentration of VEGF protein in supernatant and real-time fluorescence quantitative PCR method was used to detect the expression level of VEGF MRNA. The maximum absorbance of ARPE-19 cells was 1.063 鹵0.031 in 100 渭 mol 路L ~ (-1) ARPE-19 cells treated with CoCl _ (2). The results of CCK-8 screening showed that the cell proliferation rate was 94.58 鹵3.80% in the Compactopril group (20 mg 路L ~ (-1) 路L ~ (-1)). The cell proliferation rate of 20 渭 mol 路L ~ (-1) PDGF receptor inhibitor group was 96.72 鹵5.44, and the effect was better than that of other corresponding groups. This concentration was selected for follow-up experiments. The results of factorial analysis showed that each drug treatment group (Compactopril PDGF-receptor inhibitor, Compactopril combined with PDGF receptor inhibitor) was compared with the hypoxia group after hypoxia injury. The cell proliferation activity decreased, the cell migration decreased and the concentration of PDGF protein decreased, especially in the Compactopril combined with PDGF receptor inhibitor group. Compared with hypoxia group, the expression of VEGF mRNA in ARPE-19 cells in Compactopril group and combined treatment group was lower than that in hypoxia group. Conclusion the proliferation and migration of ARPE-19 cells can be induced by hypoxia injury. The relative expression of VEGF protein was increased and the expression of VEGF mRNA was up-regulated. Compactopril combined with PDGF receptor inhibitor can inhibit the proliferation, migration and the expression of VEGF mRNA in ARPE-19 cells after hypoxia injury. The effect was better than that of Compathep alone.
【作者單位】： 徐州醫(yī)科大學研究生學院;徐州醫(yī)科大學附屬醫(yī)院眼科;
【分類號】：R774
【正文快照】： 眼部新生血管性疾病的共同病理改變是病理性1.2方法新生血管形成。研究表明,細胞內(nèi)穩(wěn)定的氧含量是1.2.1 ARPE-19細胞的培養(yǎng)采用含體積分數(shù)細胞生存的關(guān)鍵,在缺血、缺氧、炎癥等因素刺激下,10%胎牛血清的DMEM/F12培養(yǎng)基在37℃、體積分脈絡(luò)膜毛細血管血液灌注不良,視網(wǎng)膜色素上

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