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目的:观察低氧诱导因子-1α(Hypoxia inducible factor-1α,HIF-1α)在化学诱导的低氧状态下培养的原代鼠胚成骨细胞中的表达变化,并初步探讨在低氧条件下HIF-1α对成骨细胞VEGF表达的影响及与血管生成的关系。方法:在无菌条件下取妊娠19天左右的KM胎鼠颅骨,进行原代鼠胚成骨细胞的培养,并对细胞进行纯化及鉴定。将终浓度为125umol/L的氯化钴加入成骨细胞培养液中模拟缺氧状态,并设培养不同时间(0,3,6,9,12小时),采用反转录聚合酶链反应(RT-PCR)技术检测低氧培养不同时间的成骨细胞中HIF-1α和VEGF的表达情况。结果:成骨细胞在低氧培养后,其HIF-1αmRNA和VEGFmRNA的表达水平逐渐增加,在9小时达到高峰,随后下降,但仍高于低氧处理前水平。结论:HIF-1α可能通过调节VEGF表达水平而促进血管生成,并且这种促进作用可能和缺氧程度有关。
OBJECTIVE: To observe the expression changes of Hypoxia inducible factor-1α (HIF-1α) in primary murine embryo osteoblasts cultured under chemically induced hypoxia condition and to investigate the effects of hypoxia inducible factor-1α Effect of HIF-1α on VEGF expression in osteoblasts and its relationship with angiogenesis. Methods: Under the condition of aseptic conditions, the fetus of KM embryos about 19 days gestation was used for the culture of primary mouse embryo osteoblasts. The cells were purified and identified. The final concentration of 125umol / L of cobalt chloride into osteoblast culture medium simulated hypoxia, and set the culture at different times (0,3,6,9,12 hours), using reverse transcription polymerase chain reaction RT-PCR) was used to detect the expression of HIF-1α and VEGF in osteoblasts cultured in hypoxia for different time. Results: After hypoxia, the expression of HIF-1αmRNA and VEGF mRNA increased gradually and reached a peak at 9 hours, then decreased, but still higher than that before hypoxia treatment. Conclusion: HIF-1α may promote angiogenesis by regulating the expression of VEGF, and this promotion may be related to the degree of hypoxia.