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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 of bone marrow mesenchymal stem cells (BMSCs) intervention on hyperoxia-induced lung injury in neonatal rats. Methods SD rat BMSCs were isolated and cultured from 4 to 6 weeks in vitro. BMSCs were transfected with lentivirus carrying enhanced green fluorescent protein gene, and hyperoxia-induced lung injury was induced in neonatal SD neonatal rats at 3 days of age by hyperoxia exposure for 7 days. In the intervention group, 1 × 105 BMSCs (0.1ml) were injected into the caudal vein in the intervention group and 0.1ml phosphate buffered saline (PBS) in the control group. At 3, 7 and 14 days after injection, radial alveolar count (RAC) in lung tissue of rats was measured by histopathology. The expression of NF-κB and PECAM-1 were detected by immunohistochemistry. expression. Results Compared with hyperoxia group, the RAC value of BMSCs intervention group was significantly higher than that of hyperoxia group [(9.4 ± 0.5) vs (7.4 ± 0.4), (12.6 ± 0.5) vs 9.6 ± 0.4, P <0.05] , The levels of NF-κB at 3, 7 and 14 days after intervention were significantly lower than those in hyperoxia group [(10.0 ± 0.5) vs (14.5 ± 0.4), (6.0 ± 0.4) vs (7.8 ± 0.2) vs (P <0.05), while the PECAM-1 level was significantly higher than that in the hyperoxia group [(11.3 ± 0.4) vs (10.1 ± 0.3), (12.3 ± 0.8) (P <0.05). The levels of NF-κB and PECAM-1 in the hyperoxia group were similar to those in the air group [(5.8 ± 0.6) vs (5.6 ± 0.4) vs 17.2 ± 0.3) (17.5 ± 0.3), P> 0.05]. Conclusion BMSCs express green fluorescent protein after lentiviral transfection, which is suitable for in vivo tracing. BMSCs injected through the tail vein can reduce the hyperoxia-induced lung injury in neonatal rats, which may be related to the inhibition of NF-κB activation and the increase of PECAM-1 expression by BMSCs.