沉默非肌肉肌球蛋白ⅡA的骨髓间充质干细胞对烟雾吸入性损伤大鼠早期肺损伤的影响

Effects of non-muscle myosin Ⅱ A silenced bone marrow mesenchymai stem cells on lung damage of rats at early stage of smoke inhalation injury

目的探讨沉默非肌肉肌球蛋白ⅡA（NMⅡA）的骨髓间充质干细胞（BMSC）对烟雾吸入性损伤大鼠早期肺损伤的影响。方法将40只SD大鼠按照完全随机法分为对照组10只、单纯损伤组10只、BMSC组10只和NMⅡA－BMSC组10只。对照组大鼠正常吸入空气，其余3组大鼠吸入烟雾制备烟雾吸入性损伤模型。伤后30 min，单纯损伤组大鼠尾静脉注射1 mL生理盐水，BMSC组大鼠注射1 mL 1×107个/mL的第5代BMSC，NMⅡA－BMSC组大鼠注射1 mL 1×107个/mL沉默NMⅡA的BMSC。伤后24 h，取各组大鼠腹主动脉血和右肺，血气分析仪检测PaO2、PaCO2和pH值，干湿质量法测定肺湿干质量比，HE染色观察肺病理变化；收集肺泡灌洗液（BALF），ELISA法检测TNF－α、IL－10含量。对数据行单因素方差分析、Kruskal－Wallis H检验、LSD检验。结果（1）伤后24 h，与对照组比较，单纯损伤组、BMSC组、NMⅡA－BMSC组大鼠PaO2明显降低（P值均小于0.05），PaCO2明显升高（P值均小于0.05）。与单纯损伤组比较，BMSC组、NMⅡA－BMSC组大鼠PaO2明显升高（P值均小于0.05），PaCO2明显降低（P值均小于0.05）。与BMSC组比较，NMⅡA－BMSC组大鼠PaO2明显升高（P〈0.05）。单纯损伤组大鼠动脉血pH值较对照组明显降低（P〈0.05）。（2）伤后24 h，对照组、单纯损伤组、BMSC组和NMⅡA－BMSC组大鼠肺湿干质量比分别为4.36±0.15、7.79±0.42、5.77±0.18、5.11±0.20。与对照组比较，其余3组大鼠肺湿干质量比明显升高（P值均小于0.05）。与单纯损伤组比较，BMSC组、NMⅡA－BMSC组大鼠肺湿干质量比明显降低（P值均小于0.05）。与BMSC组比较，NMⅡA－BMSC组大鼠肺湿干质量比明显降低（P〈0.05）。（3）伤后24 h，对照组大鼠肺泡结构完整，无异常。与单纯损伤组比较，BMSC组及NMⅡA－BMSC组大鼠肺损伤明显减轻，肺泡结构相对完整，肺泡壁未增厚，炎性细胞浸润明显减轻。（4）伤后24 h，与对照组比较，单纯损伤组、BMSC组大鼠BALF中TNF－α含量明显升高（P值均小于0.05）。与单纯损伤组比较，BMSC组和NMⅡA－BMSC组大鼠BALF中TNF－α含量明显降低（P值均小于0.05）。与对照组比较，单纯损伤组、BMSC组、NMⅡA－BMSC组大鼠BALF中IL－10含量明显升高（P值均小于0.05）。与单纯损伤组比较，BMSC组和NMⅡA－BMSC组大鼠BALF中IL－10含量明显升高（P值均小于0.05）。与BMSC组比较，NMⅡA－BMSC组大鼠BALF中IL－10含量明显升高（P〈0.05）。结论沉默NMⅡA的BMSC能够减轻烟雾吸入性损伤大鼠早期肺损伤，比单纯使用BMSC更为有效。

正文作者信息基金 7 关键词 5English Abstract评论阅读 15 评论 0相关资源论文 ｜ 视频版权归中华医学会所有. 未经授权,不得转载、摘编本刊文章,不得使用本刊的版式设计. ObjectiveTo investigate the effects of non-muscle myosin ⅡA （NMⅡA） silenced bone marrow mesenchymal stem cells （BMSCs） on the lung damage of rats at early stage of smoke inhalation injury.MethodsForty Sprague-Dawley rats were divided into control, simple injury, NMⅡA-BMSCs, and BMSCs groups according to the completely random method, with 10 rats in each group. Rats in control group inhaled air normally, while rats in the latter 3 groups inhaled smoke to reproduce model of smoke inhalation injury. At 30 min post injury, rats in simple injury group were injected with 1 mL normal saline via caudal vein, and rats in group BMSCs were injected with 1 mL the fifth passage of BMSCs （1×107/mL）, and rats in group NMⅡA-BMSCs were injected with 1 mL NMⅡA silenced BMSCs （1×107/mL）. At post injury hour （PIH） 24, abdominal aorta blood and right lung of rats in each group were harvested, and then arterial partial pressure of oxygen （PaO2）, arterial partial pressure of carbon dioxide （PaCO2）, and pH value were detected by blood gas analyzer. Ratio of wet to dry weight of lung was determined by dry-wet weight method. Pathological changes of lung were observed with HE staining. Bronchoalveolar lavage fluid （BALF） were collected, and then tumor necrotic factor-α （TNF-α） and interleukin-10 （IL-10） content of BALF was determined by enzyme-linked immunosorbent assay. Data were processed with one-way analysis of variance, Kruskal-Wallis H test, and least-significant difference test.Results（1） At PIH 24, compared with those in control group, PaO2 values of rats in simple injury, BMSCs, and NMⅡA-BMSCs groups were obviously decreased （with P values below 0.05）, and PaCO2 values were obviously increased （with P values below 0.05）. Compared with those in simple injury group, PaO2 values of rats in groups NMⅡA-BMSCs and BMSCs were obviously increased （with P values below 0.05）, while PaCO2 values were obviously decreased （with P values below 0.05）. PaO2 value of rats in group NMⅡA-BMSCs was obviously increased as compared with that in group BMSCs （P〈0.05）. The pH value of arterial blood of rats in simple injury group was obviously lower than that in control group （P〈0.05）. （2） At PIH 24, ratios of wet to dry weight of lung of rats in control, simple injury, BMSCs, and NMⅡA-BMSCs groups were 4.36±0.15, 7.79±0.42, 5.77±0.18, and 5.11±0.20, respectively. Compared with that in control group, ratio of wet to dry weight of lung of rats was obviously increased in the other 3 groups （with P values below 0.05）. Compared with that in simple injury group, ratio of wet to dry weight of lung of rats was obviously decreased in groups BMSCs and NMⅡA-BMSCs （with P values below 0.05）. Compared with that in group BMSCs, ratio of wet to dry weight of lung of rats in group NMⅡA-BMSCs was obviously decreased （P〈0.05）. （3） At PIH 24, alveolar structure of rats in control group was complete without abnormality. Compared with those in simple injury group, lung injury and infiltration of inflammatory cells of rats in groups BMSCs and NMⅡA-BMSCs were obviously alleviated, and alveolar structure was relatively complete with no thickening of alveolar wall. （4） At PIH 24, compared with that in control group, TNF-α content of BALF of rats in simple injury and BMSCs groups was obviously increased （with P values below 0.05）. Compared with that in simple injury group, TNF-α content of BALF in groups BMSCs and NMⅡA-BMSCs was obviously decreased （with P values below 0.05）. Compared with that in control group, IL-10 content of BALF in simple injury, NMⅡA-BMSCs and BMSCs groups were obviously increased （with P values below 0.05）. Compared with that in simple injury group, IL-10 content of BALF in groups BMSCs and NMⅡA-BMSCs was obviously increased （with P values below 0.05）. Compared with that in group BMSCs, IL-10 content of BALF in group NMⅡA-BMSCs was obviously increased （P〈0.05）.ConclusionsNMⅡA silenced BMSCs can alleviate lung damage of rats at early stage of smoke inhalation injury, showing better effectiveness than using BMSCs only.