孟鲁司特联合骨髓间充质干细胞移植治疗脊髓损伤模型大鼠  

Effect of montelukast combined with bone marrow mesenchymal stem cell transplantation on spinal cord injury in rat models

作　　者：孙建威 杨新明[2] 张瑛[2] Sun Jianwei;Yang Xinming;Zhang Ying(Graduate School of Hebei North University,Zhangjiakou 075000,Hebei Province,China;Department of Orthopedics,The First Affiliated Hospital of Hebei North University,Zhangjiakou 075000,Hebei Province,China)

机构地区：[1]河北北方学院研究生学院,河北省张家口市075000 [2]河北北方学院附属第一医院骨科,河北省张家口市075000

出　　处：《中国组织工程研究》2021年第25期3962-3969,共8页Chinese Journal of Tissue Engineering Research

基　　金：河北省卫生厅2011年医学科学研究重点项目计划(20110176),项目负责人:杨新明;2013年度河北北方学院创新人才培育基金项目(CXRC1322),项目负责人:杨新明。

摘　　要：背景:联合多种治疗手段能够针对不同靶点进行有效干预,发挥协同或叠加效应,已成为当今临床治疗脊髓损伤的新理念。目的:探讨孟鲁司特联合骨髓间充质干细胞移植对脊髓损伤大鼠神经功能恢复的作用。方法:60只8周龄健康雄性SD大鼠随机分为4组,每组15只,分别为模型组、孟鲁司特组、骨髓间充质干细胞组、联合组。各组均按照Allen法进行脊髓损伤造模,在造模成功后1,6 h,通过灌胃给予孟鲁司特5 mg/kg或等量生理盐水,之后1次/d,连用30 d,造模后2 h在脊髓损伤区域一次性注入1μL BrdU标记的骨髓间充质干细胞悬液或等量PBS。造模后1,3,7,14,21,30 d,应用BBB评分和Rivlin斜板实验进行大鼠后肢运动功能评价;造模后7 d,采用苏木精-伊红染色和Nissl染色观察大鼠脊髓组织病理学变化;采用免疫组化法检测大鼠脊髓组织胶质纤维酸性蛋白、白细胞介素1β的表达以及BrdU阳性标记率;采用Western blot法检测大鼠脊髓组织中凋亡相关蛋白Bcl-2、Bax和Caspase-3的表达。结果与结论:①造模后7,14,21,30 d,联合组、孟鲁司特组、骨髓间充质干细胞组BBB评分和Rivlin斜板实验结果明显好于模型组(P <0.05);②造模后7 d,联合组、孟鲁司特组、骨髓间充质干细胞组炎性反应及水肿减轻,损伤空洞面积和胶质瘢痕面积减小,损害节段结构改善;③造模后7 d,联合组、孟鲁司特组、骨髓间充质干细胞组胶质纤维酸性蛋白、白细胞介素1β表达明显低于模型组(P <0.05),联合组BrdU阳性率高于骨髓间充质干细胞组(P=0.000);④造模后7 d,联合组、孟鲁司特组、骨髓间充质干细胞组Bax和Caspase-3的表达显著低于模型组(P <0.05),Bcl-2的表达显著高于模型组(P <0.05);⑤联合组上述所有指标均优于孟鲁司特组、骨髓间充质干细胞组;⑥结果表明,孟鲁司特联合骨髓间充质干细胞移植能够显著改善脊髓损伤大鼠神经运动功能,其机制�BACKGROUND:Combination with a variety of treatment methods can effectively intervene for different targets,play a synergistic or superimposed effect,and has become a new concept of clinical treatment of spinal cord injury.OBJECTIVE:To investigate the effect of montelukast combined with bone marrow mesenchymal stem cell transplantation on the recovery of neurological function in rats with spinal cord injury.METHODS:Sixty 8-week-old healthy male Sprague-Dawley rats were randomly divided into four groups,each with 15 rats,namely model group,montelukast group,bone marrow mesenchymal stem cell group,and combination group.The spinal cord injury model was performed in each group according to the Allen method,and montelukast 5 mg/kg or the equivalent amount of normal saline was administered by gavage at 1 and 6 hours after the model was successfully established,and then once a day for 30 days.At 2 hours after modeling,1μL of BrdU-labeled bone marrow mesenchymal stem cell suspension or an equivalent amount of PBS was injected into the spinal cord injury area at a time.At 1,3,7,14,21,and 30 days after modeling,the BBB score and Rivlin inclined plate test were used to evaluate the motor function of the rat’s hind limbs.At 7 days after modeling,the histopathological changes of spinal cord were observed by hematoxylin-eosin staining and Nissl staining.Immunohistochemical method was applied to detect the expression of glial fibrillary acidic protein,interleukin-1βand BrdU positive labeling rate in rat spinal cord tissue.Western blot assay was used to detect the expression of apoptosis-related proteins Bcl-2,Bax and Caspase-3 in rat spinal cord tissue.RESULTS AND CONCLUSION:(1)At 7,14,21,and 30 days after modeling,the BBB score and Rivlin inclined plate test results of the combination group,montelukast group and bone marrow mesenchymal stem cell group were significantly higher than those of the model group(P<0.05).(2)At 7 days,,the inflammatory reaction and edema were reduced;the area of injury cavity and glial scar was red

关 键 词：干细胞 骨髓间充质干细胞 脊髓损伤 孟鲁司特 胶质纤维酸性蛋白 白细胞介素1Β BrdU染色 

分 类 号：R496[医药卫生—康复医学] R318[医药卫生—临床医学]