摘要
目的:初步探讨低氧环境对人牙周膜成纤维细胞(periodontal ligament cells,PDLCs)增殖与凋亡的影响,以及低氧诱导因子-1α(hypoxia inducible factor-1α,HIF-1α)在该过程中的作用。方法:体外常氧条件和低氧(1%O2)条件下培养PDLCs,采用四甲基偶氮唑盐(MTT)法检测低氧诱导前后PDLCs生长速度的差异;流式细胞术比较PDLCs凋亡水平的变化。Western免疫印迹和实时定量PCR检测HIF-1α的表达水平。通过小干扰RNA(HIF1α-Si)转染PDLCs,检测低氧诱导下HIF-1α水平、细胞活性以及细胞凋亡水平的变化。结果:人PDLCs在低氧环境中培养48h后细胞活性显著抑制,凋亡水平增高,H IF-1α在蛋白和m R NA水平均显著升高。小干扰RNA(si RNA)转染PDLCs并于低氧下培养后HIF-1α表达明显降低,同时细胞活性增加、细胞凋亡显著下降。结论:低氧能通过上调HIF-1α表达抑制PDLCs增殖并促进其凋亡。
Objective： To investigate the effect of hypoxia on proliferation and apoptosis in human periodontal ligament cells （PDLCs）, and the role of HIF-1α in this process. Methods： PDLCs were cultured under hypoxia （1%O2） or normoxia. MTT was used to detect cell viability of PDLCs treat with or without hypoxia. Effect of hypoxia on apoptosis of PDLCs was detected by Annexin V-APC on a flow cytometer. Western blot and qRT-PCR were used to detect expression of HIF-1α at protein and mRNA level respectively. HIF-1 （HIF1） was transfected into PDLCs by small interfering RNA. HIF-1α, cell viability, as well as apoptosis, were furthered detected in PDLCs. Results： In the hypoxic environment, the activity of PDLCs was significantly inhibited after 48 h of culture. The level of apoptosis was increased, and the levels of HIF-1 protein and mRNA were significantly increased After knocking down of HIF-1α with siRNA, HIF-1α was significantly downregulated in PDLCs under hypoxia, as well as increased cell viability but decreased apoptosis. Conclusion： Hypoxia inhibits proliferation and promotes apoptosis of PDLCs via upregulated HIF-1α.
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出处
《中华老年口腔医学杂志》
2017年第3期170-174,共5页
Chinese Journal of Geriatric Dentistry
基金
国家自然科学基金青年基金项目(项目编号:81600899)
