小剂量雌激素肌肉注射联合骨髓间充质干细胞移植修复脊髓损伤  

Combined use of low-dose 17-beta estradiol and bone marrow mesenchymal stem cell transplantation for spinal cord repair

作　　者：康聪[1] 孟宪勇[2] 杨新明[2] 成垚昱 张振梁[1] Kang Cong;Meng Xian-yong;Yang Xin-ming;Cheng Yao-yu;Zhang Zhen-liang(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,河北省张家口市075000 [2]河北北方学院附属第一医院,河北省张家口市075000

出　　处：《中国组织工程研究》2017年第25期4022-4027,共6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):BMSC体外分化移植治疗脊髓损伤临床与基础研究~~

摘　　要：背景:多种手段联合应用可针对不同靶点进行有效干预,发挥叠加或协同作用,已成为当前脊髓损伤治疗的新思路。目的:探讨骨髓间充质干细胞移植联合应用小剂量雌激素对大鼠脊髓损伤后神经功能恢复及炎症反应的影响。方法:将70只雄性SD大鼠随机分5组,假手术组(n=10)仅暴露脊髓,不做损伤处理;模型组、雌激素组、干细胞组、联合组(每组15只)采用改良的Allen’s法建立脊髓损伤模型。干细胞组、联合组造模后立即在损伤部位移植骨髓间充质干细胞;雌激素组、联合组造模后第1,24小时肌肉注射17β-雌二醇。造模后第1,3,5,7天,进行BBB行为学评价;造模后6,12,24,72 h,应用ELISA法测定脊髓匀浆组织白细胞介素1β、肿瘤坏死因子α水平,TUNEL染色观察脊髓组织切片神经细胞凋亡情况。结果与结论:(1)与假手术组比较,模型组造模后不同时间点的BBB行为学评分下降(P<0.05);与模型组比较,干细胞、雌激素组、联合组造模后5,7 d的BBB行为学评分升高(P<0.05),且联合组高于干细胞、雌激素组(P<0.05);(2)与假手术组比较,模型组造模后6,12,24 h的白细胞介素1β、肿瘤坏死因子α水平升高(P<0.05);与模型组比较,干细胞、雌激素组、联合组造模后12,24 h的白细胞介素1β、肿瘤坏死因子α水平降低(P<0.05),且联合组低于干细胞、雌激素组(P<0.05);(3)造模后72 h,模型组神经细胞凋亡率高于假手术组,干细胞、雌激素组、联合组低于模型组(P<0.05),且以联合组最低(P<0.05);(4)结果表明在脊髓损伤中,雌激素与骨髓间充质干细胞联合应用具有协同作用,能更有效抑制神经细胞凋亡,促进脊髓功能恢复。BACKGROUND:Combined use of multiple interventions for different targets play superimposed or synergistic effects,which has become the current idea for spinal cord injury treatment.OBJECTIVE:To investigate the synergistic effects of low doses of17-βestradiol combined with bone marrow mesenchymal stem cells(BMSCs)transplantation on the recovery of motor function and inflammatory reactions after spinal cord injury in rats.METHODS:The10of70male Sprague-Dawley rats served as sham group in which the spinal cord was only exposed but with no treatment,and the rest60rats were used to make animal models of spinal cord injury using modified Allen’s method and then randomized into four groups(n=15per group):model,estrogen,stem cell and combined treatment groups.Rats in the stem cell and combined treatment groups were given BMSCs transplantation at injured side;rats in the estrogen and combined treatment groups were given intramuscular injection of17-βestradiol at1and24hours after modeling.At1,3,5and7days after modeling,rat functional recovery was evaluated by the Basso,Beatlie,Bresnahan score.The expressions of interleukin-1βand tumor necrosis factor-αin the injured spinal cord were detected by ELISA at6,12,24,and72hours after modeling.Apoptosis in nerve cells was observed using TUNEL staining.RESULTS AND CONCLUSION:The Basso,Beatlie,Bresnahan scores were declined significantly after modeling,increased at5and7days after stem cell transplantation,estrogen treatment or their combined treatment(P<0.05),especially in the combined treatment group(P<0.05).The levels of interleukin-1βand tumor necrosis factor-αwere elevated gradually after spinal cord injury(P<0.05),but the levels decreased significantly at12and24hours in stem cell,estrogen and combined treatment groups(P<0.05),and this decrease trend was more significant in the combined treatment group compared with the stem cell and estrogen groups(P<0.05).At72hours after modeling,the rate of TUNEL positive cells was highest in the model group(P<0.05)and lowest in the com

关 键 词：干细胞 移植 脊髓损伤 雌激素 骨髓间充质干细胞 白细胞介素1Β 肿瘤坏死因子α 细胞凋亡 细胞移植 

分 类 号：R394.2[医药卫生—医学遗传学]