Intrathecal transplantation of bone marrow stromal cells attenuates blood-spinal cord barrier disruption induced by spinal cord ischemia-reperfusion injury in rabbits
Intrathecal transplantation of bone marrow stromal cells attenuates blood-spinal cord barrier disruption induced by spinal cord ischemia-reperfusion injury in rabbits
复制标题
DOI：
10.1016/j.jvs.2012.11.087
复制
发表时间：
2013-10-01
影响因子：
4.3
通讯作者：
Ma, Hong
中科院分区：
文献类型：
--
作者：
Fang, Bo;Wang, He;Ma, Hong
目的：脊髓鞘内灌注骨髓基质细胞对脊髓缺血再灌注损伤有一定的治疗作用。血脊髓屏障对维持脊髓内稳态和神经功能至关重要。然而，骨髓基质细胞在脊髓缺血再灌注损伤后对血脊髓屏障的作用尚不清楚。本研究探讨骨髓基质细胞在脊髓缺血再灌注损伤所致血脊髓屏障破坏中的作用及其可能机制。方法：本研究是在中国医科大学第一附属医院中心实验室进行的前瞻性动物研究。本研究选用日本大白兔81只（体重1.8 ~ 2.6 kg）。采用兔肾下主动脉阻断30分钟诱导脊髓缺血再灌注损伤。在诱导损伤2天前，鞘内注射骨髓间质细胞（1 × 10(8)个，0.2 ml磷酸盐缓冲盐水）。采用Tarlov标准评估后肢运动功能，采用组织学检查计数腹侧灰质运动神经元。采用Evans蓝（EB）和硝酸镧作为血管示踪剂检测血脊髓屏障的通透性。采用Western blot、实时聚合酶链反应和免疫荧光分析检测紧密连接蛋白occludin的表达和定位。测定基质金属蛋白酶-9 （MMP-9）和肿瘤坏死因子- α (tnf - α)的表达。结果：脊髓缺血再灌注损伤后4小时（5.41 +/- 0.40 vs 7.94 +/- 0.36 μ g/g, P < 0.01）和24小时（9.03 +/- 0.44 vs 15.77 +/- 0.89 μ g/g, P < 0.01），骨髓基质细胞鞘内移植可减轻神经运动功能障碍和组织病理学缺损（P < 0.01），减轻EB外渗。此外，骨髓基质细胞治疗抑制脊髓缺血再灌注损伤引起的occludin降低（P < 0.01）。骨髓基质细胞减少了MMP-9和tnf - α的过度表达（P < 0.01）。结论：骨髓基质细胞鞘内预先移植稳定了兔短暂性主动脉闭塞模型脊髓缺血再灌注损伤后血脊髓屏障的完整性。这种有益效果部分是由抑制MMP-9和tnf - α介导的，代表了减轻主动脉闭塞后脊髓损伤的潜在治疗方法。
Objective: Intrathecal administration of bone marrow stromal cells has been found to produce beneficial effects on ischemia-reperfusion injury to the spinal cord. The blood-spinal cord barrier is critical to maintain spinal cord homeostasis and neurologic function. However, the effects of bone marrow stromal cells on the blood-spinal cord barrier after spinal cord ischemia-reperfusion injury are not well understood. This study investigated the effects and possible mechanisms of bone marrow stromal cells on blood-spinal cord barrier disruption induced by spinal cord ischemia-reperfusion injury.Methods: This was a prospective animal study conducted at the Central Laboratory of the First Affiliated Hospital, China Medical University. The study used 81 Japanese white rabbits (weight, 1.8-2.6 kg). Spinal cord ischemia-reperfusion injury was induced in rabbits by infrarenal aortic occlusion for 30 minutes. Two days before the injury was induced, bone marrow stromal cells (1 x 10(8) in 0.2-mL phosphate-buffered saline) were transplanted by intrathecal injection. Hind-limb motor function was assessed using Tarlov criteria, and motor neurons in the ventral gray matter were counted by histologic examination. The permeability of the blood-spinal cord barrier was examined using Evans blue (EB) and lanthanum nitrate as vascular tracers. The expression and localization of tight junction protein occludin were assessed by Western blot, real-time polymerase chain reaction, and immunofluorescence analysis. Matrix metalloproteinase-9 (MMP-9) and tumor necrosis factor-alpha (TNF-alpha) expression were also measured.Results: Intrathecal transplantation of bone marrow stromal cells minimized the neuromotor dysfunction and histopathologic deficits (P < .01) and attenuated EB extravasation at 4 hours (5.41 +/- 0.40 vs 7.94 +/- 0.36 mu g/g; P < .01) and 24 hours (9.03 +/- 0.44 vs 15.77 +/- 0.89 mu g/g; P < .01) after spinal cord ischemia-reperfusion injury. In addition, bone marrow stromal cells treatment suppressed spinal cord ischemia-reperfusion injury-induced decreases in occludin (P < .01). Finally, bone marrow stromal cells reduced the excessive expression of MMP-9 and TNF-alpha (P < .01).Conclusions: Pre-emptive intrathecal transplantation of bone marrow stromal cells stabilized the blood-spinal cord barrier integrity after spinal cord ischemia-reperfusion injury in a rabbit model of transient aortic occlusion. This beneficial effect was partly mediated by inhibition of MMP-9 and TNF-alpha and represents a potential therapeutic approach to mitigating spinal cord injury after aortic occlusion.