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目的观察小鼠角膜碱烧伤后修复过程中凝血酶敏感蛋白2(TSP2)的表达情况。方法制作小鼠角膜碱烧伤模型(实验组,35只)。采用HE染色、免疫组织化学和逆转录聚合酶链反应(RT-PCR)等方法,观察对照组(正常小鼠5只)及实验组小鼠碱烧伤后3、6、12、24、48、96、192h(每时相点5只)角膜组织的形态学改变和TSP2、tsp2的表达情况。结果HE染色显示,实验组小鼠伤后24h起角膜内有明显的血管生成。免疫组织化学和RT-PCR检测结果一致显示,TSP2及tsp2在对照组及实验组小鼠角膜组织中均有表达,多见于上皮层的基底层。与对照组的TSP2表达水平(0.48±0.15)比较,实验组小鼠伤后3h TSP2的表达水平即上升(P<0.05),即6h达第1个峰值1.54±0.45,24h降至低谷0.73±0.19,随后又回升,96h时达第2个峰值1.79±0.63,此后下调,至伤后192h降至对照组水平(P>0.05).tsp2的表达趋势与之相似。结论小鼠角膜碱烧伤后TSP2的表达量随时相点的变化呈波动的趋势,这一变化可能属于应激条件下的代偿性增高。
Objective To observe the expression of thrombospondin 2 (TSP2) during the repair of cornea alkali burns in mice. Methods Mouse corneal alkali burn model (experimental group, 35). Using HE staining, immunohistochemistry and reverse transcription-polymerase chain reaction (RT-PCR) and other methods to observe the control group (normal mice 5) and experimental mice after alkali burn 3,6,12,24,48, The morphological changes of corneal tissue and the expression of TSP2 and tsp2 were observed at 96 and 192h (5 points at each time point). Results HE staining showed that there was obvious angiogenesis in the cornea from 24h after injury in experimental group. The results of immunohistochemistry and RT-PCR showed that both TSP2 and tsp2 were expressed in the corneal tissue of the control group and the experimental group, which was more common in the basal layer of the epithelium. Compared with control group, the expression level of TSP2 increased 3h after injury (P <0.05), ie, peaked at 1.54 ± 0.45 at 6h and decreased to 0.73 ± 0.19, and then rose again, reached the second peak at 96h 1.79 ± 0.63, then down to 192h after injury to the control group (P> 0.05) .tsp2 expression trend similar. Conclusion The expression of TSP2 in corneal alkali burn mice showed a fluctuating trend at any time point. This change may be compensatory increase under stress conditions.