中文摘要：

为研究低氧诱导因子1α（HIF1α）及其下游基因胰岛素样生长因子2（IGF-2）和血管内皮生长因子（VEGF）在注射孕马血清促性腺激素（PMSG）（促排组）的促排卵巢与注射生理盐水（对照组）的普通卵巢组织中的mRNA转录水平,构建小鼠HIF1α基因真核表达载体,观察其转染的小鼠颗粒细胞中HIF1α基因的表达情况,以探讨HIF1α与小鼠卵泡发育的关系。用荧光定量PCR检测不同处理组织中HIF1α、VEGF及IGF-2 mRNA转录水平,免疫组化技术定位HIF1α在卵泡中的表达,基因重组技术构建HIF1α-pcDNA3.1真核表达载体并将其转染小鼠颗粒细胞,荧光定量PCR与Western blot技术检测转染细胞中HIF1α表达情况。结果表明：HIF1α在促排小鼠卵巢中的mRNA转录水平极显著高于普通小鼠卵巢组织（P〈0.01）,在促排组和对照组的其他组织内HIF1αmRNA转录水平并无显著差异;HIF1下游基因VEGF和IGF-2在促排小鼠卵巢组织中的转录水平分别显著（P〈0.05）与极显著（P〈0.01）高于普通小鼠卵巢组织中的转录水平;卵泡发育至有腔卵泡后,HIF1α才开始在卵泡中表达;HIF1α-pcDNA3.1真核表达载体能够在颗粒细胞中转录HIF1αmRNA并翻译成蛋白。结论：HIF1α只在有腔卵泡内表达,其转录水平与卵巢内有腔卵泡数量成正相关,HIF1α通过上调VEGF和IGF-2参与卵泡的发育及成熟。

英文摘要：

The study was aimed to investigate HIF1α mRNA transcriptional levels in different tissues of mice injected pregnant mare serum gonadotropin（ PMSG） or injected normal saline,confirm the changes of HIF1α mRNA transcriptional levels only occurred in superovulated ovaries,detect the transcriptional levels of insulin-like growth factor 2（ IGF-2） and vascular endothelial growth factor（ VEGF） in superovulated ovaries and general ovaries,construct vectors of HIF1α to transfect into mouse granulosa cells and identify its expression,and explore the relationship between HIF1α and mouse follicle development,so as to offer references for the revealing of the molecular mechanism in follicular development process. Real-time PCR was employed to examine the transcriptional levels of HIF1αmRNA in ovary,kidney,liver,brain,spleen of mice injected PMSG or injected normal saline. The same way was used in IGF-2 and VEGF,which were HIF1 target genes. The expression of HIF1α in the antral follicles was confirmed by immunohistochemical techniques. HIF1α-pcDNA3. 1 expression vectors were constructed with recombinant DNA technology. Liposome method was used for transfection of cells. Western blot and real-time PCR were used to examine the protein expression or mRNA transcriptional levels of HIF1α. The results showed that the mRNA transcriptional levels of HIF1α and its target genes（ VEGF and IGF-2） were higher in superovulated ovaries then in general ovaries. HIF1α only expressed in antral follicles. The constructed HIF1α-pcDNA3. 1 plasmid could transcribe HIF1α mRNA and express HIF1α protein efficiently in mouse granulosa cells. Conclusions： The expression of HIF1α only occurred in antral follicles,and the transcriptional levels of HIF1α mRNA depend on the number of antral follicles in ovaries. The high expression of HIF1α in superovulated ovaries indicated that HIF1α probablely participated in regulating the process of follicular development and oocytes maturation,and this regulation was achieved through