| 李 燕,田 铸,徐梦林,翟 军,谢百发.缺氧对大鼠脑微血管内皮细胞增殖和凋亡的影响及其机制研究[J].,2019,19(2):222-226 |
| 缺氧对大鼠脑微血管内皮细胞增殖和凋亡的影响及其机制研究 |
| Effect and Mechanism of Hypoxic Micro-environment on the Proliferation and Apoptosis of Rat Brain Vascular Endothelial Cells |
| 投稿时间：2018-04-07 修订日期：2018-04-30 |
| DOI：10.13241/j.cnki.pmb.2019.02.005 |
| 中文关键词: 缺血性脑卒中 缺氧 低氧诱导因子-1α 增殖 凋亡 |
| 英文关键词: Cerebral arterial thrombosis, Hypoxia, HIF-1α, Proliferation, Apoptosis |
| 基金项目:国家自然科学基金项目（81071537） |
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| 中文摘要: |
| 摘要 目的：探讨缺氧对大鼠脑微血管内皮细胞增殖和凋亡的影响及其可能的分子机制。方法：从2周龄的SD大鼠中提取脑微血管内皮细胞。体外模拟脑缺氧微环境，将体外培养的脑微血管内皮细胞分别置于常氧(21% O2)和低氧环境(1% O2)下处理6、12和24小时，采用四甲基偶氮唑盐(MTT)法观测不同时间点细胞增殖能力的变化；用AnnexinV-FITC/PI双标流式细胞术观察不同时间点细胞凋亡情况；Real time-PCR和Western blot法检测细胞中(Hypoxia-inducible factor-1α) HIF-1α mRNA和蛋白的表达。进一步使用HIF-1α siRNA靶向沉默HIF-1α基因，再检测低氧环境下HIF-1α mRNA和蛋白表达、细胞增殖以及凋亡水平的变化。结果：随着低氧处理时间的延长，大鼠脑微血管内皮细胞的增殖能力被显著抑制，同时凋亡水平显著增加，HIF-1α mRNA和蛋白表达水平显著升高。使用HIF-1α siRNA特异性阻断HIF-1α表达后，低氧环境下HIF-1α mRNA和蛋白表达明显降低，同时细胞活性增加，细胞凋亡率显著下降。结论：缺氧微环境能够通过上调HIF-1α的表达抑制大鼠脑微血管内皮细胞增殖并促进其凋亡。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the effect and mechanism of hypoxia on the proliferation and apoptosis of rat brain vascular endothelial cells. Methods: Rat brain vascular endothelial cells were isolated from 2 weeks old SD rat. Cells were treated under hypoxia (1% O2) or normoxia(21%O2) condition for 6, 12 and 24 hours. MTT assay was performed to detect the cell proliferation rate. Annex- inV-FITC/PI flow cytometry was used to monitor the apoptosis rate of rat brain vascular endothelial cells. The mRNA and protein expres- sion levels of HIF-1α and α-catenin were detected by RT-PCR and Western blot. Furthermore, HIF-1α siRNA was used to investigate the role of HIF-1α on hypoxia-mediated proliferation and invasion of rat brain vascular endothelial cells. Results: The results shown that hy- poxia could inhibit the proliferation and induce the apoptosis of rat brain vascular endothelial cells in a time-dependent manner. The mRNA and protein level of HIF-1α was upregulated by hypoxia treatment. HIF-1α siRNA could significantly reverse hypoxia-mediated prolifer- ation and apoptosis of rat brain vascular endothelial cells. Conclusion: Hypoxia inhibits the proliferation and promotes apoptosis of rat brain vascular endothelial cells through upregulation of HIF-1α expression. |
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