论文部分内容阅读
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。
OBJECTIVE: To detect the expression of hypoxia inducible factor-1α (HIF-1α) and vascular endothelial growth factor (VEGF) in human periodontal ligament cells (hPDLCs) ) Of the expression. METHODS: Human periodontal ligament cells were cultured by tissue block method. The third generation cells were used in the experiment. The cells were divided into two groups at 20% O2, 5% CO2, 75% N2 (control group) and 1% O2, % N2 (hypoxic group) in 37 ℃ incubator for 24h and 48h. The expression of HIF-1α and VEGF were detected by RT-PCR and immunohistochemistry. SPSS 13.0 software package for t test, one-way ANOVA and LSD test. Results: The expression of HIF-1αmRNA in hypoxia group was not significantly different from that in control group. The expression of VEGF mRNA in hypoxia 24h and 48h was significantly higher than that in control group (P <0.05). Immunocytochemistry showed that HIF-1α staining was positive in hypoxia group and a large number of HIF-1α protein was accumulated in the nucleus, and the expression of HIF-1α protein increased with time of hypoxia, but not in normoxia group The expression of VEGF increased in a time-dependent manner, and the expression of VEGF in hypoxia group was significantly stronger than that in control group. After 48 hours of normoxia, VEGF was weakly positive expression. Conclusion: Hypoxia can alter the metabolic pathway of human periodontal ligament cells and up-regulate the expression of HIF-1α and related factors.