KLF7联合脂肪源性干细胞对脊髓损伤大鼠运动功能的影响及其机制  

Effect of KLF7 and adipose-derived stem cells on motor function after spinal cord injury in rats

作　　者：李文媛[1] 王莹[1] 郭素芬[1] 刘星[1] 李智刚[2] 刘贵波[1] 许志鹏[1] 张际菲[1] 

机构地区：[1]牡丹江医学院,黑龙江牡丹江157011 [2]牡丹江医学院红旗医院

出　　处：《山东医药》2017年第44期5-8,共4页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(81371362);黑龙江省科学基金资助项目(LC2017040);黑龙江省卫生厅科研课题(2016-357)

摘　　要：目的观察KLF样转录因子7(KLF7)联合脂肪源性干细胞(ADSC)对脊髓损伤大鼠运动功能的影响,并探讨其可能的机制。方法将30只SD大鼠随机分为KLF7联合ADSC组、ADSC组及对照组,每组10只,均采用离断右半脊髓的方法制备脊髓损伤模型。KLF7联合ADSC组于脊髓损伤组织注入100μL ADSC(ADSC均采用PKH26进行标记)及2μL AAV2-KLF7腺病毒,ADSC组仅注入100μL ADSC,对照组仅注入100μL PBS。术后1天及1、2、3、4周行运动功能评分(BBB评分);术后4周,应用诱发电位仪和磁刺激器经颅磁刺激运动诱发神经电生理反应,记录神经传导速度、潜伏期及波幅。术后4周处死,采用Western boltting法检测脊髓损伤组织KLF7、神经生长因子(NGF)及其受体酪氨酸激酶A(TrkA)蛋白相对表达量,计算脊髓损伤面积百分比及ADSC数量(以PKH26的光密度值表示)。结果与术后1天比较,各组术后1、2、3、4周BBB评分均升高(P均<0.05)。术后4周,KLF7联合ADSC组与ADSC组BBB评分、神经传导速度、波幅、ADSC数量及脊髓损伤组织NGF、TrkA蛋白相对表达量均高于对照组,潜伏期及脊髓损伤面积百分比均低于对照组,且KLF7联合ADSC组上述变化更明显(P均<0.05)。KLF7联合ADSC组脊髓损伤组织KLF7蛋白相对表达量均高于ADSC组及对照组(P均<0.05)。结论KLF7联合ADSC可减轻脊髓损伤大鼠的神经损伤,改善其运动功能;其机制可能与KLF7促进ADSC存活及提高脊髓损伤组织NGF、TrkA表达有关。Objective To observe the effect of KLF7 and adipose-derived stem cell （ADSC） on motor function after spinal cord injury （SCI） in rats, and to probe the possible mechanism. Methods Thirty healthy SD rats were randomly divided into the KLF7 ＋ ADSC group, ADSC group, and control group, with 10 in each. SCI models were prepared by the method of transection of right half spinal cord. The 100 μL ADSC （ADSC labeled with PKH26） and 2 μL AAV2-KLF7 were injected into the SCI tissues in the KLF7 ＋ ADSC group, 100 μL ADSC was injected in the ADSC group, and 100 μL PBS was injected in the control group. At 1 day and 1, 2, 3, 4 weeks after operation, BBB was used to evaluate rats＇ double lower limb functional recovery in each group. At week 4 after the operation, evoked electrophysiological responses were evoked by evoke potential instrument and tcMMEP, and the nerve conduction velocity, latency and amplitude were re- corded. The rats were killed at 4 weeks after the operation, KLFT, NGF and TrkA protein in the SCI tissues were evaluated by Western blotting. The lesion area percentage of SCI and the number of ADSC were evaluated （ the number of ADSC was represented by the light density value of PKH26）. Results The BBB scores increased at 1,2, 3 and 4 weeks after opera- tion as compared with that at 1 day after operation in each group （P 〈 0.05 ）. At 4 weeks after operation, compared with the control group, BBB score, nerve conduction velocity, amplitude, the number of ADSC, NGF and TrkA protein expression in the SCI tissues increased in the KLF7 ＋ ADSC group and ADSC group, however, the latency and the percentage of lesion area decreased, moreover, the effect of KLF7 ＋ ADSC group was more significant than that of the ADSC group （P 〈0.05）. KLF7 relative protein expression in the SCI tissues in the KLF7 ＋ ADSC group was higher than that of the ADSC group and control group （ all P 〈 0.05 ）. Conclusion KLF7 combined with ADSC could improve the motor function and alleviate the nerve

关 键 词：脊髓损伤 KLF样转录因子7 脂肪源性干细胞 神经生长因子 酪氨酸激酶A 大鼠 

分 类 号：R496[医药卫生—康复医学]