hMASP2 DNA纳米脂质体对BCG感染小鼠肺组织T细胞亚群和AKT/pAKT蛋白表达的影响  

Effects of hMASP2 DNA nano-liposomes on T-lymphocyte subsets and expression of AKT/pAKT protein in lung tissue of BCG-infected mice

作　　者：高琪 张国超[1] 何琦 董信芳[1] 雒艳萍[1] 车团结 马兴铭[1] GAO Qi;ZHANG Guo-Chao;HE Qi;DONG Xin-Fang;LUO Yan-Ping;CHE Tuan-Jie;MA Xing-Ming(Department of Immunology, School of Basic Medical Science,Lanzhou University;Gansu Key Laboratory of Functional Genomics and Molecular Diagnosis, Lanzhou 730000, China)

机构地区：[1]兰州大学基础医学院 [2]甘肃省功能基因组与分子诊断重点实验室,甘肃兰州730000

出　　处：《转化医学电子杂志》2017年第10期35-39,共5页E-Journal of Translational Medicine

基　　金：兰州大学中央高校基本科研业务费专项资金(lzujbky-2017-117);兰州市科技计划项目(2016-3-107)

摘　　要：目的:构建hMASP2 DNA纳米脂质体复合物,探讨hMASP2 DNA纳米脂质体对BCG感染小鼠肺组织T细胞亚群和AKT/pAKT蛋白表达的影响.方法:构建pcDNA3.1-hMASP2重组表达质粒,以脂质体作为载体,制备质粒DNA纳米脂质体复合物并检测Zeta电位和粒径;经鼻滴入1×10~9CFU BCG 100μL,建立结核分枝杆菌感染小鼠模型,实验分DNA纳米脂质体组(pcDNA3.1-hMASP2组)和对照组,在感染当天小鼠尾静脉分别注射pcDNA3.1-hMASP2 200μL(25μg DNA)和PBS 200μL,每3天注射一次,连续治疗三周后处死小鼠;分离肺组织淋巴细胞,流式细胞仪检测T细胞亚群;肺组织免疫组化分析AKT、pAKT蛋白表达.结果:pcDNA3.1-hMASP2 DNA纳米脂质体复合物Zeta电位52.6 m V、平均粒径279.9 nm;与对照组相比,pcDNA3.1-hMASP2组小鼠肺组织CD4^+T细胞亚群百分比明显升高(P<0.05)、CD4^+PD-1^+T细胞亚群百分比降低(P<0.05);pcDNA3.1-hMASP2组小鼠肺组织AKT和pAKT的阳性细胞百分比明显增高(P<0.05).结论:构建稳定的pcDNA3.1-hMASP2 DNA纳米脂质体复合物,pcDNA3.1-hMASP2纳米脂质体增加BCG感染小鼠的肺组织的CD4^+T细胞、促进肺组织pAKT蛋白的表达,降低肺组织的PD-1^+T细胞亚群,从而上调小鼠T细胞功能.AIM: To evaluate the potential impacts of human MASP2( hMASP2) DNA nano liposomes on T-lymphocyte subsets and AKT/pAKT protein expression in lung tissue of BCG-infected mice through constructing the hMASP2 DNA nano-liposomes complex. METHODS: The pcDNA3.1-hMASP2 recombinant plasmids were constructed and then the hMASP2 DNA nanoliposomes complex were prepared with a carrier of liposome. The zeta potential and particle size of hMASP2 DNA nano-liposomes were detected. After infection with 100 μL of 1 × 10~9 CFU BCG,mice were randomly divided into two groups and received the following treatments: the mice of hMASP2 DNA nano-liposomes group were injected by tail vein with 200 μL( 25 μg DNA) of DNA nano-liposomes and the mice of control group were received PBS( 200 μL) by tail vein. All mice were injected every three days and treated for 3 weeks,then were killed. Lymphocytes were isolated from lung tissue and PD-1^+ T cell subsets were detected by flow cytometry. The expression of AKT/pAKT protein was detected with immunohistochemical staining. RESULTS: The zeta potential of pcDNA3. 1-hMASP2 DNA nano-liposome complex was 52. 6 m V and the average particle size was 279. 9 nm.Compared with the control group,the percentage of CD4^+ T-lymphocyte subsets in the lung tissue of mice treated with hMASP2 DNA nano-liposomes were significantly increased( P < 0. 05) andthe percentage of CD4^+PD-1^+ T-lymphocyte subsets were significantly decreased( P < 0. 05). The percentage of AKT and pAKT positive cells in hMASP2 DNA nano-liposomes group were significantly increased( P < 0. 05). CONCLUSION: A stable pcDNA3.1-hMASP2 DNA nano-liposome complex is firstly constructed,which shows the positive efficacy in increasing the number of CD4^+T cells,reducing the number of PD-1^+ T-lymphocyte subsets,and promoting the expression of pAKT of BCG-infected mice. These findings provide experimental evidence that hMASP2 DNA nano-liposomes display a potential role of up-regulating T-cell-mediated immunity in tuberculosis.

关 键 词：纳米脂质体 MASP2 结核病 PD-1 AKT 

分 类 号：R730.45[医药卫生—肿瘤]