Ding Rui, Lin Chunnan, Wei ShanShan, Zhang Naichong, Tang Liangang, Lin Yumao, Chen Zhijun, Xie Teng, Chen XiaoWei, Feng Yu, Wu LiHua
Department of Neurosurgery, Jingmen No. 1 People's Hospital, Jingmen 448000, Hubei, China.
Department of Neurosurgery, Maoming People's Hospital, Maoming 525000, Guangdong, China.
Mol Cells. 2017 Feb;40(2):133-142. doi: 10.14348/molcells.2017.2251. Epub 2017 Feb 13.
Previous studies have shown that bone marrow mesenchymal stromal cell (MSC) transplantation significantly improves the recovery of neurological function in a rat model of intracerebral hemorrhage. Potential repair mechanisms involve anti-inflammation, anti-apoptosis and angiogenesis. However, few studies have focused on the effects of MSCs on inducible nitric oxide synthase (iNOS) expression and subsequent peroxynitrite formation after hypertensive intracerebral hemorrhage (HICH). In this study, MSCs were transplanted intracerebrally into rats 6 hours after HICH. The modified neurological severity score and the modified limb placing test were used to measure behavioral outcomes. Blood-brain barrier disruption and neuronal loss were measured by zonula occludens-1 (ZO-1) and neuronal nucleus (NeuN) expression, respectively. Concomitant edema formation was evaluated by H&E staining and brain water content. The effect of MSCs treatment on neuroinflammation was analyzed by immunohistochemical analysis or polymerase chain reaction of CD68, Iba1, iNOS expression and subsequent peroxynitrite formation, and by an enzyme-linked immunosorbent assay of pro-inflammatory factors (IL-1β and TNF-α). The MSCs-treated HICH group showed better performance on behavioral scores and lower brain water content compared to controls. Moreover, the MSC injection increased NeuN and ZO-1 expression measured by immunochemistry/immunofluorescence. Furthermore, MSCs reduced not only levels of CD68, Iba1 and pro-inflammatory factors, but it also inhibited iNOS expression and peroxynitrite formation in perihematomal regions. The results suggest that intracerebral administration of MSCs accelerates neurological function recovery in HICH rats. This may result from the ability of MSCs to suppress inflammation, at least in part, by inhibiting iNOS expression and subsequent peroxynitrite formation.
先前的研究表明,骨髓间充质基质细胞(MSC)移植可显著改善脑出血大鼠模型的神经功能恢复。潜在的修复机制包括抗炎、抗凋亡和血管生成。然而,很少有研究关注MSC对高血压性脑出血(HICH)后诱导型一氧化氮合酶(iNOS)表达及随后过氧亚硝酸盐形成的影响。在本研究中,于HICH后6小时将MSC脑内移植到大鼠体内。采用改良神经功能缺损评分和改良肢体放置试验来测量行为结果。分别通过紧密连接蛋白-1(ZO-1)和神经元细胞核(NeuN)表达来测量血脑屏障破坏和神经元丢失。通过苏木精-伊红(H&E)染色和脑含水量评估伴随的水肿形成。通过免疫组织化学分析或CD68、离子钙结合衔接分子1(Iba1)、iNOS表达及随后过氧亚硝酸盐形成的聚合酶链反应,以及通过促炎因子(白细胞介素-1β和肿瘤坏死因子-α)的酶联免疫吸附测定来分析MSC治疗对神经炎症的影响。与对照组相比,MSC治疗的HICH组在行为评分上表现更好,脑含水量更低。此外,通过免疫化学/免疫荧光测量,MSC注射增加了NeuN和ZO-1表达。此外,MSC不仅降低了CD68、Iba1和促炎因子的水平,还抑制了血肿周围区域的iNOS表达和过氧亚硝酸盐形成。结果表明,脑内给予MSC可加速HICH大鼠的神经功能恢复。这可能至少部分是由于MSC具有通过抑制iNOS表达及随后的过氧亚硝酸盐形成来抑制炎症的能力。