【作者】 董宏彬；

【导师】 尹昭云；

【作者基本信息】 中国人民解放军军事医学科学院 ， 劳动卫生与环境卫生学， 2005， 博士

【摘要】 藏族是对高原低氧环境适应最好的民族,研究藏族人组织细胞的基因表达特性对于揭示低氧适应机制具有重要理论意义,对于寻找加速高原低氧习服措施也具有指导意义。 本研究以藏汉族骨髓组织为材料,采用抑制削减杂交技术和基因芯片技术对藏汉族骨髓组织差异表达基因进行筛选,利用定量RT-PCR方法对部分基因进行验证,应用生物信息学方法对差异表达基因进行分析。 对部分藏汉族骨髓组织差异表达基因进行验证发现,藏族人血红蛋白(Hb)基因表达与平原汉族人接近,而胚胎和胎儿期Hb基因(HbE、Hbζ和Hbγ)表达上调,促红细胞生成素受体(EPOR)的表达下调;促凋亡基因(HIFl-β、TP53、BAX)表达下调、抑制凋亡基因Bcl-2表达上调,凋亡蛋白基因Caspase2和Caspase8表达下调;细胞因子基因CCL5、IL28RA、IGFBP1与TGFBR3表达上调,参与酸碱平衡的碳酸酐酶Ⅱ(CA2)和水通道蛋白1 (AQP1)基因表达下调。 对藏汉族骨髓组织差异表达基因谱综合分析发现,HIF-1低氧反应通路基因表达下调;高氧亲和力血红蛋白组份(胚胎和胎儿期Hb基因)表达上调;凋亡抑制基因上调,促凋亡基因与凋亡蛋白基因表达下调;多数有氧代谢、无氧代谢酶类、电子传递链多种复合体、细胞周期相关蛋白以及RBC网架结构蛋白中的基因表达下调;多种细胞因子以及自由基清除酶类基因表达上调。 初步推测,HIF—1低氧反应通路基因的下调可能是藏族人低氧适应的一个特征,通过HIF-1通路基因的下调和抗凋亡能力的提高,产生细胞保护机制;藏族人通过上调高亲和力Hb基因表达,提高摄氧与供氧能力,通过降低代谢水平和增殖能力,减少氧与能量消耗,产生一种低氧条件下合理补充与利用能源的良性模式;藏族人通过下调AQP1、CA2表达,上调自由基清除酶类和一些细胞因子表达,保持细胞内环境的稳定。这种多环节、多基因的综合调控,有利于藏族人对高原低氧环境的适应。更多还原

【Abstract】 Tibetan is unique ethnic group which exhibit best adaptation to the hypoxic environment at high altitude. Investigating differential expressed genes in bone marrow between Tibetan and Han ethnic group would be helpful to explore the mechanism of Tibetan adaptation to hypoxic environment and to develop the measurements in promoting hypoxia acclimatization of sea-level residents.In order to screen the specific genes to high altitude hypoxic adaptation , the SSH cDNA library of differential expression genes in bone marrow tissue between Tibetan and Han ethnic group was constructed and sequenced, and at mean time, the expression profile of differential expression genes between Tibetan and Han was analyzed by the gene chip method by using the bone marrow tissue samples from high altitude Tibetan natives and sea-level Han ethnic group. Some differential genes were reexamined by quantity real-time RT-PCR method.The reexamined result of real-time RT-PCR suggested that Hbε, Hbζ, Hbγ , BCL2, CCL5, IL28RA, IGFBPland TGFBR3 genes were upregulated , and TP53, BAX, EPOR, AQP1, Caspase2, Caspase8 和 CA2 genes were downregulated in the Tibetan bone marrow tissue compared with Han’s.Compared with Han bone marrow myeloid tissue, genes encoded enzymes concerned with glycolysis, aerobic metabolism , genes encoded Complex I, IV, V of electron transport chain were downregulated in the Tibetan’s, furthermore cell cycle-related genes, and RBC framework structure protein genes in Tibetan’s were almost downregulated too. On the contrast, the hemoglobin genes, anti-apoptosis genes, some cytokine and chemokine genes are upregulated in the Tibetan’s.These findings indicated that the effects of high altitude hypoxia on Tibetan bone marrow may be the result of multicultural genes’ regulation. And the long-term更多还原