摘要
目的:探讨人脐带间充质干细胞移植联合小剂量超短波治疗对大鼠脊髓损伤(SCI)星形胶质细胞形成的影响。方法:将90只SD大鼠随机分成5组:假手术组(n=6)、对照组(n=21)、小剂量超短波(ultrashort wave,USW)治疗组(n=21)、人脐带间充质细胞移植组(human umbilical cord mesenchymal stem cells,hUCMSCs)(n=21)、USW治疗+hUCMSCs移植组(n=21)。hUCMSCs移植组、USW治疗+hUCMSCs移植组在损伤后立即使用微量注射器刺破硬脊膜垂直缓慢注入5μl含细胞密度为1×106/μl的细胞悬液;USW治疗组、USW治疗+hUCMSCs移植组大鼠在SCI后1d起采用对置法接受每天1次、每次7min的小剂量USW治疗。大鼠的运动功能用Basso Beattie Bresnahan评分(BBB评分)进行评定,SCI大鼠取材后分别对组织进行纵向切片行HE染色、免疫荧光染色,检测移植后hUCMSCs细胞存活及向星形胶质细胞分化,损伤区域附近内源性干细胞向星形胶质细胞分化及SCI后4周时纤维化形成的情况。结果:①SCI大鼠造模后1—2周内损伤周围组织附近,由内源性干细胞分化的星型胶质细胞形成胶质屏障,发挥保护作用。②SCI14d,移入hUCMSCs细胞位于损伤区域内,部分移入的干细胞向星形胶质细胞去分化。损伤周围的星形胶质细胞多由内源性干细胞分化而来,人源干细胞特异性标记物MAB1281在此区域不表达。③USW+hUCMSCs移植组,USW协同hUCMSCs通过旁分泌作用改善损伤区域微环境,从而促进更多内源性干细胞聚集损伤处分化为星形胶质细胞,并阻碍坏死物质及炎症扩散(P<0.01 vs对照组、小剂量USW组、hUCMSCs移植组)。④USW+hUCMSCs移植抑制SCI后巨噬细胞等细胞产生的不利于轴突生长的纤维瘢痕,hUCMSCs移植在过程中扮演重要的作用(P<0.01 vs对照组、小剂量USW组)。⑤USW+hUCMSCs移植更好促进SCI后大鼠运动恢复(P<0.01vs对照组、小剂量USW组、hUCMSCs移植组)。结论:小剂量USW治疗协同急性期人脐带间充质�
Objective:To explore the effects of human umbilical cord mesenchymal stem cell transplantation combined with low dose ultrashort wave(USW)therapy on the formation of astrocytes after spinal cord injury in rats.Method:Ninty SD rats were randomly divided into 5 groups:sham operation group(n=6),control group(n=21),USW treatment group (n=21),human umbilical cord mesenchymal stem cells(hUCMSCs) transplantationgroup (n=21),and USW+hUCMSCs transplantation group (n=21). Spinal cord contusion was inflicted using modified Allen's method without any treatment. Cell transplantation group and USW + hUCMSCs transplantationgroup were punctured with a microinjector and 5ul cell suspension with cell density of 1×106/L was injectedvertically and slowly. The control group and the low- dose USW group were punctured with a microinjectorand 5ul fresh cell culture fluid was injected vertically and slowly. In the sham operation group,only exposedthe T10 spinal cord without contusion and injection. Rats in the USW treatment group and USW+hUCMSCstransplantation group were treated with USW treatment once a day and 7min each time one day after SCI.The motor function of the rats was evaluated using BBB score. After sacrificed,SCI rats were subjected tolongitudinal sections for HE staining and IF staining to detect the survival and differentiation of hUCMSCscells after transplantation,the differentiation of endogenous stem cells into astrocytes near the injury area,andthe formation of fibrosis at 4w after SCI.Result:①Within 1-2w after modeling,the astrocytes differentiated by endogenous stem cells formed a glial barrier and played a protective role SCI rats around the injured area. ②SCI 14d,transplanted hUCMSCs cells located in the damaged area,and some transplanted stem cells differentiated into astrocytes. The astrocytesaround the injury area were mostly differentiated from endogenous stem cells, and the specific markerMAB1281 of human stem cells was not expressed in this region. ③In the USW + hUCMSCs transplantationgroup, USW combin
作者
王帅
董洪选
陈丹莹
苏浩
孙师
赵丽娜
高健
张立新
WANG Shuai;DONG Hongxu-an;CHEN Danying(Shengjing Hospital Rehabilitation Centre,China Medical University,Shenyang,Liaoning Province,110134)
出处
《中国康复医学杂志》
CAS
CSCD
北大核心
2021年第6期649-656,共8页
Chinese Journal of Rehabilitation Medicine
基金
国家自然科学青年基金资助项目(81101462)
辽宁省自然科学基金项目(201602875)
辽宁省公益科学基金项目(2016003001)
中国医科大学校培育学科建设经费资助项目(112-3110119071)。
关键词
脊髓损伤
内源性干细胞
脐带间充质细胞移植
小剂量超短波治疗
spinal cord injury
endogenous stem cell
umbilical cord mesenchymal cell transplantation
lowdose ultrashort wave therapy
