缺氧诱导因子-1α在大鼠角膜发育过程中的表达

发布时间：2018-07-21 18:09

【摘要】：目的观察大鼠角膜在胚胎期及生后发育期缺氧诱导因子-1α(hypoxia-inducible factor-1 alpha,HIF-1α)的表达,探讨HIF-1α与角膜发育之间的关系。方法取实验用Wistar大鼠分为8组:胚胎12 d(E12)、胚胎16 d(E16)、胚胎20 d(E20)、生后1 d(P1)、生后7 d(P7)、生后15 d(P15)、生后21 d(P21)、成年组,每组3只鼠6眼,共48眼。每只鼠取一眼,采用免疫组织化学方法检测HIF-1α的表达,另一眼采用实时荧光定量PCR方法检测角膜组织中HIF-1αmRNA的表达(E12、E16时鼠胚眼球较小,无法分离角膜组织,未行PCR检测)。结果免疫组织化学染色结果显示,大鼠在角膜发育过程中,HIF-1α的表达存在时空变化。E12时,由表面外胚层分化的两层细胞内可见明显棕黄色颗粒。E16时,分化形成的上皮层、基质层、内皮层中HIF-1α阳性表达显著。E20时,棕黄色颗粒集中表达在上皮层、内皮层,基质层染色较弱。P1、P7时,大鼠眼睑粘连,角膜各层中HIF-1α维持少量表达,上皮、内皮细胞较基质细胞染色明显。P15时,HIF-1α在上皮层阳性着色尤其强烈,细胞核、细胞浆棕黄色颗粒明显,基质层未见阳性着色,内皮层见少量细胞核着染。P21时,HIF-1α的阳性表达更加集中在上皮层基底细胞,基质层、内皮层表达阴性。成年组,角膜中仅在上皮基底细胞见HIF-1α阳性表达,余未见阳性。实时荧光定量PCR显示HIF-1αmRNA在角膜发育期各组表达量不同,差异有统计学意义(F=97.00、P0.05)。P1组与E20组之间,成年组与P21组之间比较,差异均无统计学意义(P=0.20、0.05),其余各组间差异均有统计学意义(均为P0.05)。结论 HIF-1α在大鼠角膜发育过程中存在阳性表达,且随发育时期不同表达部位、表达量发生变化。HIF-1α可能在角膜各层分化、细胞分裂过程中发挥重要作用。
[Abstract]:Objective to observe the expression of hypoxia-inducible factor-1 alpha-HIF-1 伪 (hypoxia-inducible factor-1 alpha-HIF-1 伪) in rat cornea at embryonic and postnatal stages, and to explore the relationship between HIF-1 伪 and corneal development. Methods Wistar rats were divided into 8 groups: embryonic 12 d (E12), embryo 16 d (E16), embryo 20 d (E20), postnatal day 1 (P1), postnatal 7 d (P7), postnatal 15 d (P15), postnatal 21 d (P21). The expression of HIF-1 伪 was detected by immunohistochemistry in each mouse, and the expression of HIF-1 伪 mRNA in cornea was detected by real-time fluorescence quantitative PCR. Results Immunohistochemical staining showed that the expression of HIF-1 伪 in the cornea of rats had temporal and spatial changes. E12, when the two layers of cells differentiated from the surface ectoderm could be seen in the brown granules. E16, the epithelium and stroma were differentiated into epithelium and stroma. When HIF-1 伪 positive expression was significant in the endodermis. E20, the brown granules were mainly expressed in the upper cortex, endodermis and stroma layer. When the staining was weak in the stromal layer, the eyelid adhesion was observed in rats, and HIF-1 伪 was expressed in a small amount in each layer of cornea, epithelium. The positive staining of HIF-1 伪 in the epithelium was stronger in endothelial cells than in stromal cells (P 15), the brown granules in nucleus and cytoplasm were obvious, and no positive staining was found in matrix layer. The positive expression of HIF-1 伪 was more concentrated in basal cells, stromal layer and endodermis of endodermis. In adult group, only HIF-1 伪 positive expression was found in epithelial basal cells, but no HIF-1 伪 expression was found in other corneas. The expression of HIF-1 伪 mRNA in each group was different in the development phase of cornea by real-time fluorescence quantitative PCR, and the difference was statistically significant (F _ (97.00) P _ (0.05). There was a significant difference between the P1 group and the E20 group, and between the adult group and the P21 group. There was no significant difference between the two groups (P < 0.05), but there were significant differences among the other groups (P0.05). Conclusion there is a positive expression of HIF-1 伪 in the development of cornea in rats. HIF-1 伪 may play an important role in the process of corneal differentiation and cell division.
【作者单位】：青岛眼科医院;
【分类号】：R77

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