腺病毒介导的Slit2及Slit2 ShRNA转染缺氧诱导的人RPE细胞对人脉络膜微血管内皮细胞增殖的影响

发布时间：2018-12-29 11:22

【摘要】：目的:观察腺病毒介导的Slit2及Slit2 ShRNA转染缺氧诱导的人视网膜色素上皮(retinal pigment epithelial cells,RPE)细胞对人脉络膜微血管内皮细胞(human choroidal microvascular endothelial cell,HCMEC)增殖的影响,探讨Slit2在脉络膜新生血管中的可能作用,为脉络膜新生血管(choroidal neovascularization,CNV)提供新的治疗思路。方法:体外培养并鉴定人RPE细胞、HCMEC;200μmol/L氯化钴建立化学缺氧模型,Transwell小室建立细胞共培养模型;将缺氧的RPE细胞随机分为Slit2组(加入Slit2)、Slit2 ShRNA组(加入Slit2 ShRNA)、空腺病毒组(加入空腺病毒)、缺氧组,12、24、48 h后采用CCK 8(Cell Counting Kit-8,CCK 8)法检测HCMEC的增殖。结果:不同组别存在组间差别,差异均有统计学意义(F=98.122,P=0.000),不同时间点存在差别(F=3388.913,P=0.000),组别与时间点的交互作用(F=82.863,P=0.000)。Slit2组吸光度(absorbance,A)值在24 h、48 h均高于其他组(与缺氧组P=0.001,其余P=0.000),Slit2 ShRNA组A值在24 h、48 h均低于其他组(48 h与缺氧组P=0.003,与空腺病毒组P=0.008,其余P=0.000)。结论:Slit2的高表达可明显促进HCMEC的增殖,沉默RPE细胞中的Slit2的表达后,会明显抑制HCMEC的增殖。
[Abstract]:Aim: to investigate the effect of adenovirus-mediated Slit2 and Slit2 ShRNA transfection on the proliferation of human retinal pigment epithelium (retinal pigment epithelial cells,RPE) cells induced by hypoxia on the proliferation of human choroidal microvascular endothelial cells (human choroidal microvascular endothelial cell,HCMEC). To explore the possible role of Slit2 in choroidal neovascularization (choroidal neovascularization,CNV). Methods: human RPE cells were cultured and identified in vitro, HCMEC;200 渭 mol/L cobalt chloride was used to establish chemical hypoxia model, and Transwell chamber was used to establish cell coculture model. The hypoxic RPE cells were randomly divided into Slit2 group (adding Slit2), Slit2 ShRNA group (adding Slit2 ShRNA), empty adenovirus group), hypoxia group, CCK 8 (Cell Counting Kit-8,CCK 8 method after 48 h. Results: there were significant differences among different groups (F _ (98.122) P ~ (0.000), different time points (F _ (3388.913) P ~ (0.000), interaction between groups and time points (F _ (82.863), P ~ (0.000). The absorbance (absorbance,A) of Slit2 group was significantly higher than that of other groups (P 0. 001, P 0. 000), and the A value of Slit2 ShRNA group was lower than that of other groups (48 h and hypoxia group P 0. 003). Compared with empty adenovirus group (P 0. 008), the rest (P 0. 000). Conclusion: the high expression of Slit2 can obviously promote the proliferation of HCMEC, but after silencing the expression of Slit2 in RPE cells, it can inhibit the proliferation of HCMEC.
【作者单位】：重庆医科大学附属第二医院眼科;
【基金】：国家自然科学基金资助项目(编号:81170858)
【分类号】：R773.4

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