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目的:研究大鼠脊髓缺血损伤后β管蛋白及其mRNA的表达变化。方法:利用β管蛋白的单抗及地高辛标记cDNA探针,借助于免疫组化、原位杂交、图像分析等方法,对腹主动脉阻断(肾动脉上)30min后再灌流不同时间点大鼠缺血损伤脊髓神经元β管蛋白及其mRNA的变化进行了定性定量研究。结果:SCI后β管蛋白及mRNA均明显降低,3d达最低水平,7d有所恢复,到14dβ管蛋白mRNA恢复到伤前,而β管蛋白仍维持低水平。结论:β管蛋白破坏,可能与Ca2+激活的中性蛋白酶有关;β管蛋白与其mRNA的分离,提示β管蛋白的减少,除转录水平减少所致翻译产物降低外,破坏增多亦是可能的原因。
Objective: To investigate the expression of β-tubulin and its mRNA after spinal cord ischemic injury in rats. Methods: The cDNA probes were labeled with β-tubulin monoclonal antibody and digoxigenin. After 30 min occlusion of the abdominal aorta (renal artery) and reperfusion at different times by means of immunohistochemistry, in situ hybridization and image analysis, The changes of β-tubulin and its mRNA in the spinal cord of rats with ischemic injury were qualitatively and quantitatively studied. Results: The expression of β-tubulin and mRNA were significantly decreased after SCI, reached the lowest level on the 3rd day, recovered on the 7th day, and returned to pre-injury on the 14th day. However, the β-tubulin remained low. Conclusion: The destruction of β-tubulin may be related to the neutral protease activated by Ca2 +. The separation of β-tubulin and its mRNA suggests that the decrease of β-tubulin may be caused by the decrease of translational products as well as the decrease of translation products .