全反式维甲酸抑制微血管内皮细胞增殖

目的：研究不同浓度的全反式维甲酸（all-trans retinoic acid,atRA）对体外培养的小鼠脑微血管内皮细胞株bEnd.3增殖的影响。 方法： 将体外培养的bEnd.3细胞随机分为五组：空白对照组（加入atRA溶剂二甲亚砜, DMSO）、10-9、10-8、10-7和10-6mol/L atRA组。分别在24 h、48 h和72 h收集细胞，通过流式细胞术、MTT法检测不同浓度的atRA对bEnd.3增殖的影响，并采用免疫细胞化学法检测bEnd.3增殖细胞核抗原（PCNA）的表达。在作用明显的时点（20 h）做小鼠血管新生相关基因芯片检测。 结果： 10-6mol/L atRA组在24 h明显抑制细胞增殖；PCNA表达明显降低；20 h时11种血管新生相关基因表达明显下调。 结论： 在体外细胞培养中，10-6mol/L 的atRA作用20-24 h 时，通过部分血管新生相关基因表达下调和PCNA的表达减少抑制bEnd.3细胞增殖。

Inhibitory effect of all-trans retinoic acid on proliferation of microvascular endothelial cells

AIM: To evaluate the effects of all-trans retinoic acid (atRA) on the proliferation in cultured mouse cerebral microvascular endothelial cells (bEnd.3) . METHODS: Cultured cells were divided into five groups randomly, one as control group, the other four groups were 10-9, 10-8, 10-7 and 10-6 mol/L group. Effects of atRA on proliferation in bEnd.3 cells were detected by flow cytometry and immunocytochemitry of PCNA and MTT at 24 h, 48 h and 72 h. The effects of atRA (10-6 mol/L group) on the expressions of angiogenic genes in bEnd.3 cells were studied using microarray. RESULTS: The results of MTT and flow cytometry showed that all- trans retinoic acid at concentration of 10-6 mol/L significantly inhibited the proliferation of bEnd.3 cells. Immunocytochemical staining showed the expression of PCNA was markedly decreased in bEnd.3 cells at 24 h after treatment with atRA. Microarray results demonstrated that there were 11 down - regulated angiogenic genes and 2 up - regulated angiogenic genes in 10-6mol/L atRA group. CONCLUSION: All - trans retinoic acid at concentration of 10-6mol/L may significantly inhibit the proliferation of bEnd.3 cells treated for 24 h ire vitro via down-regulation of angiogenic genes and PCNA expression.