Wnt-3a信号蛋白促进大鼠脊髓损伤修复作用的实验研究.docVIP

Wnt-3a信号蛋白促进大鼠脊髓损伤修复作用的实验研究 　　作者：祖波，尹宗生，张辉 作者单位：国家自然科学基金资助项目 1 246003 安徽医科大学附属安庆医院骨科 　2 230022 安徽医科大学第一附属医院骨科 　　【摘要】 目的 研究外源性Wnt-3a信号蛋白对脊髓损伤的修复作用，并探讨其作用机制。方法 取40只成年雌性SD大鼠在T10节段制备适度脊髓挫伤模型。随机从中取20只为损伤对照组(Group 1)，另外20只为损伤治疗组(Group 2)。脊髓损伤3天后用Wnt-3a蛋白治疗。这些大鼠的功能恢复通过Basso、Beattie、Bresnahan (BBB)开放视野运动评分来观察。这些大鼠分批在损伤后14天或28天被处死，取出损伤节段脊髓用来组织病理学检查，同时用5-溴脱氧尿嘧啶核苷(BrdU)和神经细胞标记物进行免疫组化染色。结果 两组大鼠在伤后一周运动功能有明显的恢复。不过，到损伤后28天，我们观察发现，损伤治疗组中的大鼠BBB评分比损伤对照组中的评分高出7.0分左右(Group 2∶16.94±1.18,Group 1∶9.89±1.29;P0.05)，光镜和电镜检查发现Wnt-3a蛋白对髓鞘形成和轴突再生有一定的修复效果。免疫组织化学发现，在损伤后2周，Wnt-3a蛋白治疗组诱导分化的神经元数目明显多于损伤对照组。结论 大鼠脊髓损伤施加外源性的Wnt-3a蛋白可以一定程度上促进轴突传导和功能恢复，并且可以促进神经元数目的增加，通过双标染色提示这些神经元来源于内源性神经干细胞。 　　【关键词】 分化;神经干细胞;大鼠;脊髓损伤;Wnt-3a蛋白 　　Repair effect of Wnt-3a protein on contused adult rat spinal cord 　　ZU Bo,YIN Zong-sheng,ZHANG Hui.Anqing Hospital,Anhui Medical University,Anqing 246003,China 　　[Abstract] Objective To evaluate the functional recovery of acute spinal cord injuried rats treated with exogenous Wnt-3a signal protein administration and to explore its mechanism.Methods Moderate spinal cord contusion injury was made in 40 adult Sprague Dawley (SD) rats at T10.Twenty rats served as contusion controls(group 1).Twenty rats were treated with Wnt-3a for three days after injury (group 2).The functional recovery of the rats was observed through Basso,Beattie,Bresnahan (BBB) open field locomotor score.Rats were killed at 14 or 28 days after injury,then spinal cords were removed for histopathological examinations,and the expression of the bromodeoxyuridine (BrdU) plus neural cell markers was stained with immunohistochemical method.Results Rats of two groups receiving a contusive injury recovered substantial function within 1 week.By 28 days,rats in groups 2 scored 7.0 points better on the BBB scores than rats in group 1 (group 2=16.94,after 28 days vs. group 1=9.89 points;P0.05).Light and electron microscopic studies showed that the Wnt-3a treated group had moderate repair effect o