ApoJ基因修饰的骨髓间充质干细胞移植治疗大鼠脑出血的作用机制  

Therapeutic effect of transplantation of ApoJ-modified bone marrow mesenchymal stem cells on intracerebral hemorrhage in rats and its action mechanism

作　　者：刘茂春 刘亮 普娟 陈慧 刘学良 徐斌 郑晓梅 LIU Mao-chun1,LIU Liang2,PU Juan1,CHEN Hui1,LIU Xue-liang2,XU Bin2,ZHENG Xiao-mei1(1.Department of Neurology,2.Department of Neurosurgery,the Affiliated Hospital of Southwest Medical University,Luzhou 646000,Sichuan,Chin)

机构地区：[1]西南医科大学附属第一医院神经内科,泸州医学硕士研究生646000 [2]西南医科大学附属第一医院神经外科,泸州646000

出　　处：《医学研究生学报》2018年第5期470-475,共6页Journal of Medical Postgraduates

基　　金：四川省教育厅重点项目课题(12ZA075);四川省教育厅课题(14110);四川省医学科研课题计划项目(S17074)

摘　　要：目的目前载脂蛋白J(Apo J)对脑出血的神经保护作用尚不明确,文中旨在探讨携带载脂蛋白J基因的骨髓间充质干细胞(BMSCs)移植对脑出血大鼠的治疗作用及其可能的机制。方法体外分离、培养大鼠BMSCs;通过脂质体介导重组质粒p EGFP-N1-Apo J转染BMSCs。将96只SD大鼠随机数字表法分为3组:Apo J/BMSCs组、BMSCs组和对照组,通过"二次注血法"制作大鼠脑出血(ICH)模型,造模成功后24 h分别移植等体积转染Apo J基因的BMSCs悬液、BMSCs悬液和等渗盐水至3组大鼠脑出血部位。在移植后1、3、5、7d的不同时间点,利用改良神经功能评分(m NSS)评估大鼠神经功能恢复情况,通过干湿重法测量脑含水量,并采用Western blot技术检测各组大鼠补体C3表达的差异。结果 3组大鼠移植后1 d的m NSS评分差异无统计学意义(P>0.05),移植后3、5、7 d时Apo J/BMSCs组的m Nss评分[(8.13±0.99、6.75±1.04、5.63±0.52)分]显著低于BMSCs组[(9.25±1.28、8.50±1.41、7.00±0.54)分]和对照组[(10.88±0.84、9.75±0.89、7.88±1.25)分],且BMSCs组显著低于对照组(P<0.05)。Apo J/BMSCs组1、3、5、7 d脑组织含水量[(78.17±0.82)%、(78.68±0.55)%、(77.00±0.58)%、(75.89±0.46)%]显著低于同时间点BMSCs组[(78.83±0.56)%、(79.12±0.26)%、(78.13±0.46)%、(76.86±0.29)%]和对照组[(80.38±0.35)%、(81.47±0.26)%、(79.74±0.41)%、(78.44±0.44)%],其中BMSCs组显著低于对照组(P<0.05)。Western blot结果显示,Apo J/BMSCs组1、3、5、7 d的C3表达(0.096±0.011、0.083±0.005、0.064±0.009、0.045±0.007)较BMSCs组(0.212±0.014、0.164±0.013、0.105±0.010、0.091±0.004)和对照组(0.440±0.006、0.604±0.011、0.333±0.010、0.141±0.003)显著下调(P<0.05),其中BMSCs组低于对照组(P<0.05)。结论 Apo J可通过抑制补体激活介导的继发性脑损伤,减轻脑水肿,从而促进脑出血大鼠神经功能的恢复。Objective The nerve-protective effect of Apolipoprotein J（ Apo J） in intracerebral hemorrhage（ ICH） is not yet clarified. This study aimed to explore the therapeutic effect of transplantation of bone marrow mesenchymal stem cells（ BMSCs） carrying the Apo J gene on intracerebral hemorrhage in rats and its possible action mechanism. Methods Rat BMSCs were isolated,cultured in vitro,and transfected with the recombinant plasmid p EGFP-N1-Apo J mediated with lipofectamine. Ninety-six SD rats were randomly divided into groups A,B and C,and ICH models were established by two-step autologous intracranial blood injection. At 24 hours after modeling,the rats in groups A,B,and C were transplanted with the same volume of Apo J-transfected BMSC suspension,BMSC suspension and normal saline,respectively. At 1,3,5 and 7 days after transplantation,the neurofunction recovery of the rats were evaluated with modified Neurological Severity Scores（ m NSS）,the brain water content measured by the dry-wet weight method,and the expression level of complement component 3（ C3） in the brain tissue detected by Western blot. Results The m NSS exhibited no statistically significant differences among the three group of rats on the 1 st day after transplantation（ P〈0.05）,but was remarkably lower in group A than in B and C on the 3 rd（ 8.13±0.99 vs 9.25±1.28 and 10.88±0.84,P〈0.05）,5 th（ 6.75± 1.04 vs 8.50± 1.41 and 9.75± 0.89,P〈0.05） and 7 th day（ 5.63±0.52 vs 7.00±0.54 and 7.88±1.25,P〈0.05）,and markedly lower in group B than in C（ P〈0.05）. The water content in the brain tissue was also significantly lower in group A than in B and C on the 1 st（ 78.17± 0.82 vs 78.83± 0.56 and80.38±0.35,P〈0.05）,3 rd（ 78.68±0.55 vs 79.12±0.26 and 81.47± 0.26,P〈0.05）,5 th（ 77.00± 0.58 vs 78.13± 0.46 and 79.74±0.41,P〈0.05） and 7 th day（ 75.89±0.46 vs 76.86±0.29 and 78.44±0.44,P〈0.05）,and remarkably lower in group B than in C（ P〈0.05）. Western blot showed that the expre

关 键 词：载脂蛋白J 骨髓间充质干细胞 脑出血 脑含水量 神经功能评分 补体3 

分 类 号：R743.34[医药卫生—神经病学与精神病学]