携带SOD基因的骨髓间充质干细胞对小鼠百草枯肺损伤的保护作用  

The protective effect of bone marrow mesenchymal stem cells carrying antioxidant gene superoxidedismutase on paraquat lung injury in mice

作　　者：刘虹[1] 丁颖威[1] 侯跃辉[1] 赵光举[1] 卢阳[1] 陈肖[1] 蔡旗旗[1] 洪广亮[1] 邱俏檬[1] 卢中秋[1] 

机构地区：[1]温州医科大学附属第一医院急诊医学中心,325000

出　　处：《中华劳动卫生职业病杂志》2016年第1期1-7,共7页Chinese Journal of Industrial Hygiene and Occupational Diseases

基　　金：浙江省自然科学基金（LZl2H26001）;浙江省省医药卫生平台研究计划重点项目（2012ZDA034）;浙江省医学创新学科建设计划（11-CX26）;浙江省中医药重点学科计划（2012-XK-A28）;浙江省“十二五”重点学科建设项目（2012-207）

摘　　要：目的通过构建百草枯（paraquat，PQ）所致的小鼠肺损伤模型，研究携带抗氧化基因超氧化物歧化酶（supemxide dismutase，s0D）的骨髓间充质干细胞对其的治疗作用及可能机制。方法构建携带有SOD基因的过表达慢病毒并转染骨髓问充质干细胞（bone marrow mesenchymal stem cells，BMscs），建立携带有抗氧化基因SOD的BMSCs体系。将100只清洁级Balb／c小鼠按照随机数字表分为对照组、PQ染毒组、BMscs治疗组、携带空载基因的BMscs治疗组（BMsc．空载体治疗组）、携带s0D基因的BMscs治疗组（BMSC-SOD治疗组）。每组20只。PQ染毒组和各治疗组腹腔注射质量分数为20％PQ溶液（25mg/kg）构建小鼠PQ中毒肺损伤模型，对照组给予等体积生理盐水；在注射PQ溶液后4h给予相应组别球后静脉注射相应的骨髓间充质干细胞体系，分别在给予细胞治疗后的3、7、14、21d处死各组小鼠取相应肺组织标本，HE染色，观察肺组织损伤情况，观察肺组织超显微结构损伤情况。留取血液标本，用化学比色法检测SOD、丙二醛（methane dicarhoxvlic aldehyde，MDA）、谷胱甘肽（glutathione，GSH）等氧化应激指标；用ELISA方法检测肿瘤坏死因子（tumor necmsis factor，TNF）-α、转化生长因子（transfornling growth factor，TGF）-β等炎症指标，用蛋白免疫印迹法（westem Blot）检测肺组织SOD的表达情况。结果与对照组比较，第3天PQ染毒组SOD蛋白表达明显降低，第21天PQ染毒组s0D蛋白表达升高；与PQ染毒组比较，第3、21天BMSCs治疗组、BMSC-空载体治疗组、BMSc-S0D治疗组SOD蛋白表达明显升高，且BMSC-s0D治疗组升高最明显，差异均有统计学意义（P〈0．05）。与对照组比较，第3、7、14、21天PQ染毒组及BMSCs治疗组、BMSC-空载体治疗组、BMSC-S0D治疗组GsH含量和S0D活力均明显降低，且均明显高于PQ染毒组，以BMSC-s0D治疗组升高最为明显，差异均�Objective To explore the possible mechanism and protective effect of BMSCs （bone mesenchymal stem cells） carrying supernxide dismutase （SOD） geue on mice with paraquat-indnced acute lung injury. Methods To establish the cell line of BMSCs bringing SOD gene, lentiviral vector bringing SOD gene was built and co-cultured with BMSCs. A total of 100 BALB/c mice were randomly divided into five groups, namely Control group, poisoning group （PQ group）, BMSCs therapy group （BMSC group）, BMSCs-Cherry therapy group （BMSC-Cherry group）, BMSCs-SOD therapy group （BMSC-SOD group ）. PQ poisoning model was produced by stomach lavaged once with 1 ml of 25 mg/kg PQ solution, and the equal volume of normal saline （NS） was given to Control group mice instead of PQ. The corresponding BMSCs therapy cell lines were delivered to mice through the tail vein of mice 4h after PQ treatment.Five mice of each group were sacrificed 3 d,7 d, 14 d and 21 days after corresponding BMSCs therapy cell lines administration,and lung tissues of mice were taken to make sections for histological analysis. The serum levels of glutathione （GSH）, malondialdehyde （MDA）, SOD, and the levels of transforming growth factor-β （TGF-β） and tumor necrosis faetor-α （TNF-α） in lung tissue were determined. The level of SOD was assayed by Westen-blot Results Compared with Control group, the early （3 days） levels of SOD protein in lung tissue of PQ group obviously decreased, and the late （21 days） levels of SOD obviously increased, while in therapy groups, that was higher than that in PQ group, and the BMSCs-SOD group showed most obvious （all P〈0.05 ）. Compared with Control group, the levels of plasma GSH and SOD of PQ group and each therapy group wae significantly lower than those in Control group,while in therapy groups, those were higher than those of PQ group, and the BMSCs-SOD group showed most obvious （all P〈0.05 ）.Compared with Control group, the level of plasma MDA, TNF-et and TGF-13 in PQ group an

关 键 词：百草枯 中毒 呼吸窘迫综合征 骨髓间充质干细胞 超氧化物歧化酶 

分 类 号：R595.4[医药卫生—内科学]