【作者】 谢中；

【导师】 康意军；

【作者基本信息】 中南大学 ， 脊柱外科， 2011， 硕士

【摘要】 目的研究骨髓间充质干细胞与促红细胞生成素联合应用对大鼠脊髓损伤修复的影响方法将80只SD大鼠随机分成手术对照组(简称对照组,n=15)、促红细胞生成素(Erythropoietin,EPO)治疗组(简称EPO组,n=15)、大鼠骨髓间充质干细胞(BMSCs)治疗组(简称BMSCs组,n=25)和BMSCs联合EPO治疗组(简称联合组,n=25)。造模后第7天EPO组经腹腔注射促红细胞生成素,而BMSCs组、联合组则经损伤脊髓局部注射BMSCs和局部注射BMSCs的同时加用EPO腹腔注射。移植后第3d、1w、3w,BMSCs组、联合组两组分别应用双标免疫荧光法检测5-溴-2脱氧尿嘧啶核苷(BrdU)标记BMSCs的神经元中丝蛋白(NF-M)的表达情况,移植后第3周各组取脊髓组织作HE病理切片染色,再用免疫组化观察神经元中丝蛋白(NF-M)和胶质纤维酸性蛋白(GFAP)的表达情况,每个时间点取5只大鼠,移植后第1d、1w、2w、3w和4w,各组选10只大鼠分别用(BassoBeattie Bresnahan, BBB)法检测大鼠神经功能的恢复情况。所得数据采用组间均数比较t检验进行统计学处理。结果1、脊髓病理切片显示联合组较BMSCs组胶质细胞增生更加活跃,炎性细胞浸润和组织水肿显著减轻。2、双标免疫荧光结果显示移植的BMSCs在宿主脊髓中存活,从第7天开始就有NF-M表达。免疫组化结果显示联合组中NF-M和GFAP阳性细胞数目均比其余组多(P<0.05)。3、治疗组脊髓损伤修复较对照组明显,神经功能BBB评分也明显提高(P<0.05)。4、联合组在不同的时间点神经功能BBB评分都较EPO组、BMSCs组高。结论1、BMSCs移植与EPO腹腔注射是治疗大鼠脊髓损伤的有效方法；2、促红细胞生成素在体内有诱导BMSCs向神经细胞分化的能力。3. BMSCs与EPO联合应用对脊髓损伤修复具有协同作用。更多还原

【Abstract】 ObjectiveTo study the effects of bone marrow-derived mesenchymal stem cells(BMSCs) transplantation combined with Erythropoietin(EPO) in-jection for repairing spinal cord injury in ratsMethodsEighty clean healthy SD rats were divided randomly into operated group (n=15), EPO injection group (n=15), BMSCs transplantation group (n=25), BMSCs transplantation in combination with EPO group (called Associated group,n=25), At the 7 th day after operation, EPO were injected to the cavum abdominis of the rats in EPO group and the BMSCs were given by local injection to the spinal cord in BMSCs group and Associated group.21 days after transplantation, the pathological changes of the injured spinal cord was observed. After transplantation 3d, 1w,3w the expressions of neurofilament-M (NF-M) in the BMSCs which were labeled by Brdu in the spinal cord tissue were examined by immun-ofluorescence technique.21 days after transplantation, the expressions of NF-M and glialfibrial lary acidic protein(GFAP) in the BMSCs were examined by immune histochemistry,at eaeh time Point five rat. Besides,10 each in check, the degree of nerve function recovew was estimated on the 1th,7th,14th,21st and 28th day after treatment by scoring according to the Basso-Beattie-Bresnahan (BBB) locomotor rating scale. The data used are the number of inter-group comparison t-test.Result1、As compared with BMSCs Group, The Associated Group showed a milder inflammatory cell infiltration with lessened tissue edema in the spinal cord and more active proliferation of gliacyte.2、Double-labeled EnvisionTM examination turned out that the transplanted BMSCs were living alively in the host’s spinal cord. From the 7th day after transplantation, the expressions of NF-M were revealing in the injured spine cord. The amounts of NF-M and GFAP positive cells in Group D was much more than that in other Groups(P<0.05)3、The recovery of spinal cord function in EPO group, BMSCs group and Associated group were better than that of operated group, showing higher BBB scores.4、The BBB scores of Associated group is higher than the BBB scores of EPO group and BMSCs group at all the dateline date.Conclusions1、The transplantation of BMSCs and EPO injection are effective ways in cure the rat’s spinal cord injury;2、EPO can induce BMSCs differentiated into neurons.3、EPO has the ability of synergizing with BMSCs to accelerate the repairing of the spinal cord from injury.更多还原