ApoJ基因修饰的BMSCs移植对脑出血大鼠C3表达的影响  

The effect of ApoJ gene-modified BMSCs transplantation on expression of C3 in rats with intracerebral hemorrhage

作　　者：刘茂春 刘亮 普娟 陈慧 徐斌 刘学良 郑晓梅 Liu Maochun;Liu Liang;Pu Juan(Dept of Neurology,West China-Guang′an Hospital,Sichuan University,Guang′an 638000;Dept of Neurosurgery;Dept of Neurology,The Affiliated Provincial Hospital of Southwest Medical University,Luzhou 646000)

机构地区：[1]四川大学华西广安医院神经内科,广安638000 [2]西南医科大学附属医院神经外科,泸州646000 [3]西南医科大学附属医院神经内科,泸州646000

出　　处：《安徽医科大学学报》2018年第9期1364-1369,共6页Acta Universitatis Medicinalis Anhui

基　　金：四川省教育厅重点项目(编号:12ZA075);四川省医学会科研课题计划(编号:S17074)

摘　　要：目的探讨载脂蛋白J(Apo J)基因修饰的骨髓间充质干细胞(BMSCs)移植对脑出血大鼠补体3(C3)表达的影响。方法采用贴壁筛选法体外培养并纯化大鼠BMSCs,利用脂质体介导重组质粒p EGFP-N1-Apo J转染BMSCs。建立大鼠尾状核脑出血模型,随机分为Apo J/BMSCs组、BMSCs组和NS组,每组24只,三组分别于建模后24 h在脑出血部位移植等体积转染Apo J基因的BMSCs悬液、BMSCs悬液、生理盐水,各组再根据移植后喂养时间不同分为1、3、5、7 d 4个亚组,每亚组6只。采用RT-PCR法和免疫组织化学染色技术分别检测血肿周围脑组织C3 mRNA和蛋白质的表达情况。结果应用全骨髓贴壁法成功分离、培养出活性及纯度较高的BMSCs,携带Apo J基因的BMSCs能够表达外源性Apo J。RT-PCR和免疫组化结果显示,与NS组和BMSCs组比较,Apo J/BMSCs组各时间点C3的mRNA和蛋白表达均明显降低(P<0.05),且BMSCs组亦低于NS组(P<0.05)。结论Apo J基因修饰的BMSCs能明显下调C3的表达,证实Apo J对脑出血的神经保护作用是通过抑制补体激活而实现的。Objective To explore the effect of apolipoprotein J（ Apo J） gene modified bone marrow mesenchymal stem cells（ BMSCs） transplantation treatment on the expressons of complement 3（ C3） in rats with intracerebral hemorrhage. Methods Using stick wall screening method cultivates and purifies the BMSCs of rats in vitro. Recombinant plasmid p EGFP-N1-Apo J was transferred into BMSCs with lipofectamine. The rat intracerebral hemorrhage（ ICH） models was established,and then ICH rats were randomly divided into Apo J/BMSCs group,BMSCs group and NS group with 24 rats. The three groups were injected with the same volue of transfected cell suspension,BMSCs suspension and normal saline in 24 hours after modeling by intracranial injection. Each group were divided into 1 d,3 d,5 d and 7 d four subgroups according to different refeeding time,6 rats of each subgroups. The expression of C3 mRNA and protein was observed by RT-PCR and immunohistochemistry. Results Applying the bone marrow adherent method separates and develops BMSCs with high activity and purity successfully. BMSCs with Apo J gene were able to expression exogenous Apo J protein. Compared with NS group and BMSCs group,the expression of C3 mRNA and protein in Apo J/BMSCs group were significantly decreased at same time point（ P〈0. 05）,and BMSCs group was lower than NS group（ P〈0. 05）. Conclusion Apo J gene modified BMSCs transplantation can reduce expression of C3 obviously. The findings support a neuroprotective role of Apo J on ICH by inhibition of complement activation.

关 键 词：载脂蛋白J 骨髓间充质干细胞 脑出血 补体3 

分 类 号：R743.34[医药卫生—神经病学与精神病学]