rhCC10对大鼠烟雾吸入性肺损伤的作用研究

Effect of rhCC10 on smoke inhalation lung injury in rats

目的研究重组人Club细胞10-kDa蛋白(rhCC10)对大鼠烟雾吸入性肺损伤的作用。方法156只成年雄性SD大鼠随机分为对照组(sham组,n=6)、烟雾损伤组(SII组,烟雾吸入30 min,n=50)以及烟雾+rhCC10干预组[SII+rhC2组(烟雾+2 mg/kg的rhCC10)、SII+rhC10组(烟雾+10 mg/kg的rhCC10),n=50]。首先观察各组大鼠24 h存活情况,并在24 h每组随机选取6只大鼠腹主动脉血气分析后安乐死,收集支气管肺泡灌洗液(BALF)和肺组织;然后采用生化与免疫检测分析大鼠血浆、BALF及肺组织中炎症相关因子,评估BALF中蛋白渗出情况;最后通过病理学检查肺组织病理学改变。实验28 d将剩余实验大鼠安乐死,取肺组织行病理学分析。结果SII组大鼠24 h存活率为36%;SII+rhC10组存活率74%高于SII+rhC2组(62%),也高于SII组(P均<0.05);SII组血浆炎性因子和BALF中蛋白含量、炎性细胞、炎性因子水平以及肺组织髓过氧化物酶(myeloperoxidase,MPO)、高迁移率族蛋白B1(Highmobilitygroupboxl,HMGB1)蛋白表达均显著高于sham组(P均<0.05);与SII组比较,SII+rhC2、SII+rhC10组血浆炎性因子以及BALF中蛋白含量、炎性细胞、炎性因子水平均有不同程度降低(P均<0.05),其中SII+rhC2、SII+rhC10组在白细胞计数、HMGB1和BALF中蛋白水平的差异有统计学意义(P均<0.05);实验24 h SII组肺组织可见弥漫性炎性细胞浸润,肺泡内出血及大量水肿液,肺空泡腔面积较sham组显著减少(P<0.05);与SII组比较,SII+rhC2、SII+rhC10组肺炎性浸润减轻,肺空泡腔面积显著下降(P均<0.05);实验28 d SII组肺组织可见中重度肺纤维化,肺空泡腔面积较sham组明显减少(P<0.05);与SII组比较,SII+rhC2、SII+rhC10组肺纤维化减轻,肺空泡腔面积显著下降(P均<0.05);结论rhCC10可以显著提高烟雾吸入肺损伤大鼠的存活率,减轻肺部及全身炎症反应,改善肺部纤维化水平;且10 mg/kg组rhCC10效果优于2 mg/kg组。

Objective To investigate the effect of recombinant human Club 10-KDA protein(rhCC10)on smoke inhalation lung injury in rats.Methods 156 adult male SD rats were randomly divided into control group(SHAM group,n=6),smoke injury group(SII group,smoke inhalation for 30 min,n=50)and smoke+rhCC10 intervention group(SII+rhC2 group 2 mg/kg,SII+rhC10 group 10 mg/kg,n=50).First,the survival of rats in each group was observed for 24 h.Six rats were randomly selected to take the abdominal aorta for blood gas analysis before euthanasia.Bronchoalveolar lavage fluid(BALF)and lung tissue were collected.Then biochemical and immunoassay were used to analyze inflammatory factors in plasma,BALF and lung tissues of rats to evaluate protein exudation in BALF.Finally,the histopathological changes of the lung were observed through pathological examination.All experimental rats were euthanized and lung tissues were taken for pathological analysis on 28 d.Results The 24 h survival rate of rats in SII group was 36%.The survival rate of SII+rhC10 group was 74%higher than that of SII+rhC2 group and 62%higher than that of SII group(all P<0.05).Protein content,inflammatory cells,inflammatory factors,myeloperoxidase(MPO)and highmobilitygroupboxl(HMGB1)protein expression in lung tissues in SII group were significantly higher than that in SHAM group(all P<0.05).Compared with the SII group,the protein content,inflammatory cells and inflammatory factors in the plasma of the SII+rhC2 and SII+rhC10 groups were all reduced to different degrees(all P<0.05),and the differences in the WBC count,HMGB1 and BALF protein levels in the SII+rhC2 and SII+rhC10 groups were statistically significant(all P<0.05).In the 24 h experiment,diffuse inflammatory cell infiltration,alveolar hemorrhage and massive edema were observed in the lung tissues of the SII group,and the pulmonary cavity area was significantly reduced compared with that of the SHAM group(P<0.05).Compared with the SII group,the pneumonic invasion was reduced in the SII+rhC2 group and the SII+rhC10 group,and the pulmonary cavitation cavity area significantly decreased(all P<0.05).Moderate and severe pulmonary fibrosis was observed in the lung tissues of the SII group on 28 days of experiment,and the pulmonary cavitation cavity area was significantly reduced compared with the SHAM group(P<0.05).Compared with the SII group,pulmonary fibrosis was reduced in the SII+rhC2 group and the SII+rhC10 group,and the area of pulmonary cavitation cavity was significantly decreased(all P<0.05).Conclusion rhCC10 can significantly improve the survival rate of rats with smoke inhalation lung injury,reduce pulmonary and systemic inflammation,and improve the level of pulmonary fibrosis.And the rhCC10 effect in 10 mg/kg group was better than that in 2 mg/kg group.