骨髓间充质干细胞干预对高氧肺损伤新生大鼠核转录因子-κB和血小板内皮细胞黏附分子-1表达的影响  

The influence of bone marrow-derived mesenchymal stem cells on the expressions of NF-κB and PECAM-1 in neonatal rats with hyperoxia-induced lung injury

作　　者：杨旻[1] 苏浩彬[1] 张红珊[1] 

机构地区：[1]中山大学孙逸仙纪念医院儿科,广州510120

出　　处：《中国新生儿科杂志》2013年第2期122-126,共5页Chinese Journal of Neonatology

基　　金：广东省科技计划资助项目(2010B060900027)

摘　　要：目的探讨骨髓间充质干细胞(BMSCs)干预治疗新生大鼠高氧肺损伤的作用机制。方法体外分离培养4～6周龄SD大鼠BMSCs。利用携带增强型绿色荧光蛋白基因的慢病毒液对BMSCs进行转染,3日龄SD新生大鼠高氧暴露7天制备高氧肺损伤模型。建模后干预组经尾静脉注射BMSCs1×105个(0.1ml);对照组经尾静脉注射磷酸盐缓冲液(PBS)0.1ml。注射后3、7、14天,病理学方法检测大鼠肺组织放射状肺泡计数(RAC),免疫组化法检测核转录因子(NF-κB)、血小板内皮细胞黏附分子-1(PECAM-1)的表达。结果高氧干预组在BMSCs干预7、14天RAC值明显高于高氧对照组[(9.4±0.5)比(7.4±0.4),(12.6±0.5)比(9.6±0.4),P<0.05],干预3、7、14天NF-κB水平明显低于高氧对照组[(10.0±0.5)比(14.5±0.4),(6.0±0.4)比(7.8±0.2),(5.8±0.6)比(7.2±0.2),P<0.05],而PECAM-1水平明显高于高氧对照组[(11.3±0.4)比(10.1±0.3),(12.3±0.8)比(10.0±0.3),(17.2±0.3)比(10.2±0.4),P<0.05];高氧干预组在干预14天NF-κB、PECAM-1水平接近于空气干预组[(5.8±0.6)比(5.6±0.4),(17.2±0.3)比(17.5±0.3),P>0.05]。结论慢病毒转染后BMSCs表达绿色荧光蛋白,适合体内示踪。BMSCs经尾静脉注射可减轻新生大鼠高氧肺损伤,其机制可能与BMSCs抑制NF-κB活化、增加PECAM-1表达有关。Objective To investigate the mechanism underlying the therapeutical effect of bone mesenchymal stem cells （BMSCs） transplanted into neonatal rats with hyperoxia-induced lung injury. Methods BMSCs from 4-to-6-week-old Sprague-Dawley rats were isolated and cultured. Then transfected with lentivirus expressing EGFP （enhanced green fluorescent protein ）. Hyperoxia-incluced lung injury model was established in 3-day-old neonatal SD rats exposing to hyperoxia for 7 days. 1 × 10^5 BMSCs and O. 1 ml of phosphate buffered saline （PBS） were separately injected through the rats audal veins as the BMSCs transplant group and the control group. All the radial alveolar counts （RAC） were based on pathological method and the levels of NF-KB, PECAM-1 were detected using immunohistochemistry method at 1, 3, 7, 14 days after the injection. Results The RAC in BMSCs transplant group at 7 and 14-day increased significantly comparing with the control group [ （9.4 ±0. 5 ） vs. （7.4 ± 0. 4）, （ 12.6 ± O. 5 ） vs. （9.6 ± 0.4 ）, P 〈 0. 05 ]. The levels of NF-κB in the lungs at 3,7, 14-day after injection in BMSCs transplant group were significantly lower than the control group [ （ 10. 0 ±0.5） vs. （14.5±0.4）, （6.0±0.4） vs. （7.8±0.2）, （5.8±0.6） vs. （7.2±0.2）, P〈0.05]. However, the levels of PECAM-1 were higher in BMSCs transplant group [ （ 11.3 ± 0.4） vs. （ 10. 1 ± 0.3）, （12.3±0.8） vs. （10.0±0.3）, （17.2±0.3） vs. （ 10.2 ±0.4）,P〈0.05]. The levels of NF- KB and PECAM-lwere similar between the two groups at 14-day after injection [ （5.8 ±0. 6） vs. （ 5.6 ± 0. 4）, （ 17.2 ± 0.3 ） vs. （ 17.5 ± 0.3 ）, P 〉 0. 05 ]. Conclusions After transfeeted with lentivirus, BMSCs expressing GFP can be adequately traced in vivo. BMSCs can reduce hyperoxia-indueed lung injury,which may be associated with the inhibition of NF-KB and the elevation of PECAM-1 expression.

关 键 词：骨髓间充质干细胞 高氧肺损伤 核转录因子ΚB 血小板内皮细胞粘附分子-1 

分 类 号：R722.1[医药卫生—儿科]