Department of Anesthesiology, First Affiliated Hospital, China Medical University, Shenyang, Liaoning, P. R. China.
J Vasc Surg. 2013 Oct;58(4):1043-52. doi: 10.1016/j.jvs.2012.11.087. Epub 2013 Mar 7.
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.
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 × 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-α (TNF-α) expression were also measured.
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 μg/g; P < .01) and 24 hours (9.03 ± 0.44 vs 15.77 ± 0.89 μg/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-α (P < .01).
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-α and represents a potential therapeutic approach to mitigating spinal cord injury after aortic occlusion.
Clinical thoracoabdominal aorta surgery may trigger spinal cord ischemia-reperfusion injury, resulting in paraplegia as well as bladder, bowel, and sexual dysfunction. Transplantation of bone marrow stromal cells has attracted increasing attention in the field of nervous system protection, but its mechanisms have not been elucidated completely. The blood-spinal cord barrier plays a crucial role to maintain normal spinal cord function. This study suggested that intrathecal transplantation of bone marrow stromal cells stabilized blood-spinal cord barrier integrity through inhibiting the upregulation of matrix metalloproteinase-9 and tumor necrosis factor-a and ameliorated spinal cord ischemia-reperfusion injury. This may provide a novel train of thought to enhance the protective effects of bone marrow stromal cells on spinal cord injury.
骨髓基质细胞鞘内给药已被发现对脊髓缺血再灌注损伤产生有益影响。血脊髓屏障对于维持脊髓内环境稳定和神经功能至关重要。然而，骨髓基质细胞对脊髓缺血再灌注损伤后血脊髓屏障的影响尚不清楚。本研究旨在探讨骨髓基质细胞对脊髓缺血再灌注损伤诱导的血脊髓屏障破坏的作用及其可能机制。
这是在中国医科大学第一附属医院中心实验室进行的一项前瞻性动物研究。研究使用了 81 只日本白兔（体重 1.8-2.6 千克）。通过腹主动脉阻断 30 分钟诱导脊髓缺血再灌注损伤。在损伤诱导前 2 天，通过鞘内注射移植骨髓基质细胞（1×10（8）个，在 0.2-mL 磷酸盐缓冲液中）。采用 Tarlov 标准评估后肢运动功能，并通过组织学检查计数腹侧灰质中的运动神经元。采用 Evans 蓝（EB）和硝酸镧作为血管示踪剂检测血脊髓屏障通透性。通过 Western blot、实时聚合酶链反应和免疫荧光分析评估紧密连接蛋白闭合蛋白的表达和定位。还测量了基质金属蛋白酶-9（MMP-9）和肿瘤坏死因子-α（TNF-α）的表达。
鞘内移植骨髓基质细胞最小化了神经运动功能障碍和组织病理学缺陷（P<.01），并在脊髓缺血再灌注后 4 小时（5.41±0.40 比 7.94±0.36 μg/g；P<.01）和 24 小时（9.03±0.44 比 15.77±0.89 μg/g；P<.01）时减轻了 EB 外渗。此外，骨髓基质细胞治疗抑制了脊髓缺血再灌注损伤诱导的闭合蛋白减少（P<.01）。最后，骨髓基质细胞减少了 MMP-9 和 TNF-α的过度表达（P<.01）。
在短暂腹主动脉阻断兔模型中，预先鞘内移植骨髓基质细胞稳定了脊髓缺血再灌注损伤后的血脊髓屏障完整性。这种有益作用部分是通过抑制 MMP-9 和 TNF-α的表达来介导的，这代表了一种减轻主动脉阻断后脊髓损伤的潜在治疗方法。
胸腹部主动脉手术可能引发脊髓缺血再灌注损伤，导致截瘫以及膀胱、肠道和性功能障碍。骨髓基质细胞的移植在神经系统保护领域引起了越来越多的关注，但其机制尚未完全阐明。血脊髓屏障对于维持正常的脊髓功能至关重要。本研究表明，鞘内移植骨髓基质细胞通过抑制基质金属蛋白酶-9 和肿瘤坏死因子-α的上调稳定血脊髓屏障的完整性，并改善脊髓缺血再灌注损伤。这可能为增强骨髓基质细胞对脊髓损伤的保护作用提供新的思路。
