乏氧对人牙周膜细胞HIF-1α及VEGF表达的影响  

Effect of hypoxia on the expression of HIF-1α and VEGF in human periodontal ligament cells in vitro

作　　者：侯超[1] 唐开亮[1] 杨丕山[1,2] 张盼盼[1] 孙静[1] 李纾[1,2] 

机构地区：[1]山东大学口腔医院牙周科,山东济南250012 [2]山东省口腔生物医学重点实验室,山东济南250012

出　　处：《上海口腔医学》2010年第3期329-334,共6页Shanghai Journal of Stomatology

摘　　要：目的:检测人牙周膜细胞(human periodontal ligament cells,hPDLCs)在乏氧环境下乏氧诱导因子-1α(hypoxia inducible factor-1α,HIF-1α)和血管内皮生长因子(vascular endothelial growth factor,VEGF)的表达情况。方法:组织块法培养人牙周膜细胞,第3代细胞用于实验,分2组分别在20%O2,5%CO2,75%N2(对照组)和1%O2,5%CO2、94%N2(乏氧组)的37℃培养箱中培养24h和48h。采用RT-PCR技术和免疫组化技术检测HIF-1α和VEGF的表达情况。应用SPSS13.0软件包进行t检验、单因素方差分析及LSD检验。结果:乏氧组HIF-1αmRNA的表达与对照组相比,差别无统计学意义。乏氧24h与48h VEGF mRNA的表达显著高于对照组(P<0.05)。免疫细胞化学细胞爬片结果显示,乏氧组HIF-1α染色阳性,大量HIF-1α蛋白在胞核积聚,且表达量随乏氧时间增加而增加,而常氧组未观察到HIF-1α的表达;VEGF的表达呈时间依赖性增加,乏氧组VEGF的表达明显强于对照组。常氧48h后,VEGF呈微弱阳性表达。结论:乏氧可以改变人牙周膜细胞的代谢途径,上调HIF-1α及相关因子的表达。PURPOSE： To investigate the effect of hypoxia on the expression of HIF-1α and VEGF in human periodontal ligament cells （hPDLCs） in vitro. METHODS： HPDLCs were cultured in Dulbecco’s modified Eagle’s medium（DMEM） and subcultured at confluence. Experiments were carried out with the third passage of hPDLCs populations. In the normoxic control group, cells were incubated under normoxic conditions of 20%O2, 5%CO2, 75%N2 for 24 hours and 48 hours, respectively. In the hypoxic group, cells were incubated in a humidified atmosphere of 1%O2 , 5%CO2 , 94%N2 for 24 hours and 48 hours, respectively. The expression of HIF-1α and VEGF in hPDLCs in vitro was measured using reverse transcription-polymerase chain reaction. SP immunohistochemistry method was performed to localize the distribution of HIF-1α and VEGF in hPDLCs. The data was analyzed by Student’s t test, one-way ANOVA and LSD test with SPSS 13.0 software package. RESULTS： There was no significant difference between the hypoxic group and the normoxic control group in the expression of HIF-1α mRNA in hPDLCs. The expression of VEGF mRNA in the hypoxic group of 24 hours and 48 hours was both statistically higher than in the control group （P0.05）. The expression of HIF-1α protein was significantly increased with hypoxia time while the staining of HIF -1α was negative under normoxic condition. The expression of VEGF protein in hypoxic groups was significantly higher than in normoxic control groups. The staining of VEGF gradually enhanced in a time-dependent manner and was weakly positive after 48 hours under normoxic condition. CONCLUSION： Hypoxia could change the metabolic pathway of human periodontal ligament cells by upregulating the expression of HIF-1α and other relevant growth factors.

关 键 词：牙周膜细胞 乏氧 缺氧诱导因子-1Α 血管内皮生长因子 

分 类 号：R781.4[医药卫生—口腔医学]