腺病毒介导白细胞介素-6基因对脊髓星形胶质细胞生物学行为的影响  

Effect of adenovirus-mediated interleukin-6 gene on biological behavior of spinal cord astrocytes

作　　者：刘涛[1] 曹富江[1] 徐云强[1] 冯世庆[1] 

机构地区：[1]天津医科大学总医院骨科,300052

出　　处：《中华实验外科杂志》2015年第4期850-853,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金重点项目（81330042）;国家自然科学青年基金资助项目（81101362、81401784）;国家科技部对俄合作专项项目（2014DFR31210）

摘　　要：目的 观察白细胞介素-6 （IL-6）基因对体外培养的小鼠脊髓星形胶质细胞增殖、迁移、黏附的影响.方法 将携带IL-6及绿色荧光蛋白（GFP）的腺病毒载体（Ad-IL-6-GFP）及空载体（Ad-GFP）感染脊髓星形胶质细胞,实时定量聚合酶链反应（Real-time PCR）和Western blot分别检测IL-6 mRNA及胶质纤维酸性蛋白（GFAP）蛋白的表达,噻唑蓝（MTT）法检测脊髓星形胶质细胞增殖;流式细胞仪测定脊髓星形胶质细胞周期;细胞迁移实验和黏附实验检测脊髓星形胶质细胞的体外迁移和黏附能力.结果 IL-6腺病毒载体感染脊髓星形胶质细胞96 h后,Ad-IL-6-GFP组蛋白的表达量明显高于Ad-GFP组,差异有统计学意义（P＜0.01）;MTT结果显示,Ad-IL-6-GFP组吸光度（A）值明显降低,与Ad-GFP组比较,差异有统计学意义（P＜0.05）;细胞周期结果显示,IL-6基因减少了脊髓星形胶质细胞在G1期[（5.40±0.90）％]的比例,而S期[（7.26±0.40）％]和G2期[（83.52 ±0.50）％]细胞比例显著增加,与Ad-GFP组比较,差异有统计学意义（P＜0.01）;细胞迁移实验和黏附实验结果显示,IL-6载体能够促进脊髓星形胶质细胞的体外迁移和黏附能力.结论 IL-6腺病毒载体能够促进脊髓星形胶质细胞周期进程、增殖、迁移及黏附能力.Objective To investigate the effect of interleukin-6 （IL-6） gene on proliferation,migration and invasion activity of spinal cord astrocytes.Methods IL-6 gene was cloned into adenovirus vector,then the astrocytes particles expressing IL-6 were infected into astrocytes.Real-time quantitative polymerase chain reaction （Real-time PCR） and Western blotting were performed to examine the expression of IL-6 in infected astrocytes.The growth ability was detected by methyl thiazol tetrazolium （MTT） assay.The cell cycle was analyzed by flow cytometry.The adhension and migration were examined by matrix （MTX） and multi tetrazolium compounds （MTS） assays.Results Comparing to controls group and Ad-IL-6-green fluorescent protein （GFP） group,the expression levels of IL-6 mRNA and protein were both significantly up-regulated after 4 days of IL-6 adenovirus infection.Comparing with the Ad-GFP group,MTT assay showed that the A490 of Ad-IL-6-GFP group was up-regulated after adenovirus infection （P 〈 0.05）.Comparing with the Ad-GFP group,cell cycle test revealed IL-6 gene could accelerate cells in G1 [（5.40 ± 0.90） %],and the proportion of KF cells was increased in S phase [（7.26 ± 0.40） %] and G2 phase [（83.52 ± 0.50） %] after cells were infected （P 〈 0.01）.Transfection of IL-6 adenovirus stimulated the migration and invasion activity of astrocytes.Conclusion Adenovirus-mediated IL-6 gene efficiently stimulate the proliferation,cell cycle progression,migration and invasion activity of astrocytes.

关 键 词：白细胞介素-6 脊髓 星形胶质细胞 增殖 迁移 黏附 INTERLEUKIN-6 

分 类 号：R96[医药卫生—药理学]