|Alternative Title||Corneal pathological changes in mice caused by alkali burn and the expression of HIF-1α and AQP1|
|Place of Conferral||兰州|
|Keyword||昆明小鼠 角膜碱烧伤 缺氧诱导因子-1α 水通道蛋白1 细胞迁移|
角膜碱烧伤是临床上常见的眼部损伤，碱烧伤后引起角膜持续溃疡及新生血管，进而破坏角膜高度透明及无血管状态，因此一直受到关注。然而，碱烧伤后角膜伤口愈合过程仍不清楚。缺氧诱导因子-1α(hypoxia inducible factor-1α，HIF-1α) 是参与低氧反应的关键因子，在缺血性视网膜病变中能引发血管生成，水通道蛋白1（aquaporin1，AQP1）在眼睛各组织广泛表达，与眼部多种病理学过程相关。由于两者在眼部损伤病理中的重要作用，近来引起人们的普遍关注。
本研究首先对小鼠角膜碱创伤后的病理表现进行探究，其次利用免疫荧光染色和qRT-PCR技术检测HIF-1α和AQP1在角膜碱创伤后各时期的表达情况。结果显示，碱创伤后的小鼠角膜表层出现空泡，基质层纤维排布杂乱。正常组HIF-1α和AQP1分别在角膜上皮基底膜及内皮层有弱表达；碱烧伤后1 d、4 d和7 d HIF-1α mRNA表达上调，创伤后1 d和7 d AQP1 mRNA表达上调。为明确在碱性烧伤后AQP1对小鼠角膜基质细胞在伤口修复时的影响，本实验通过MTT实验及流式细胞术得到一定浓度的CoCl2用于模拟细胞缺氧环境，通过迁移实验检测AQP1对小鼠角膜基质细胞迁移能力的影响。结果显示，50 μmol/L的CoCl2可以模拟此低氧条件，此浓度模拟的低氧条件下，小鼠角膜基质细胞AQP1 mRNA的表达上调，且迁移实验结果显示低氧环境下小鼠角膜基质细胞迁移能力增强，AQP1能够促进小鼠角膜基质细胞的迁移。
综上所述，碱烧伤引起了小鼠角膜病理性改变，HIF-1α和AQP1参与了小鼠角膜碱烧伤后不同时期的病理过程，50 μmol/L的CoCl2可以作为模拟小鼠角膜基质细胞低氧条件的药物浓度，低氧条件下AQP1 的大量表达促进了角膜基质细胞的迁移，推测利于角膜伤口愈合。本实验为碱烧伤后角膜相关疾病的诊治提供了参考。
The alkali burn of corneal is a common ocular injury in clinics. It has always been concerned as it causes corneal ulcers and neovascularization, thereby destroying the highly transparent and avascular state of the cornea. So it has always been concerned. However, the process of corneal wound healing after alkali burn is still unclear. Hypoxia-inducible factor-1α (HIF-1α) is a key factor involved in hypoxia response, which can lead to angiogenesis in ischemic retinopathy. Aquaporin 1 (AQP1) is widely expressed in various tissues of the eye, and it is related to various pathological changes in the eye. It has attracted widespread attention in recent years due to the important role of HIF-1α and AQP1 in the pathology of the eye.
Initially, the corneal pathological changes by alkali burn in mice are observed. Secondly, immunofluorescence staining and qRT-PCR techniques were used to detect the expression of HIF-1α and AQP1 in different stages of corneal alkali burn. These results showed that HIF-1α in the normal group was feebly expressed in the corneal epithelial basement membrane, and AQP1 was only weakly expressed in the corneal endothelium. The expression of HIF-1α mRNA was up-regulated at 1 d, 4 d and 7 d after alkali burn, and the expression of AQP1 mRNA was up-regulated at 1 d and 7 d after injury. In order to further clarify the effect of AQP1 on mice keratocytes in wound repair after corneal alkali burns, and a certain concentration of CoCl2 was screened by MTT assay and flow cytometry to simulate the hypoxic environment of cells in this experiment. The effect of AQP1 on the migration ability of mice keratocytes was detected by cell migration assay. These results showed that given 50 μmol/L CoCl2 could be used as a hypoxia-simulating drug concentration, and the expression of AQP1 mRNA in keratocytes was up-regulated under hypoxic conditions simulated by this drug. These results showed that hypoxia induced by a certain concentration of CoCl2 could make the expression of AQP1 mRNA was up-regulated in keratocytes, and the migration ability of keratocytes was enhanced under hypoxic conditions, and AQP1 could promote the migration of keratocytes.
In summary, alkali burn could cause corneal pathological changes in mice, and HIF-1α and AQP1 are involved in the pathological process of corneal alkali burn at different stages. The 50 μmol/L CoCl2 could be used to simulating the hypoxic conditions in keratocytes. Under hypoxic conditions, the high expression of AQP1 may promote the migration of keratocytes and facilitate corneal wound healing. This experiment provides a theoretical basis for the treatment of corneal alkali burn- related diseases.
|李康. 碱烧伤致小鼠角膜病变及HIF-1α和AQP1表达的研究[D]. 兰州. 兰州大学,2018.|
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