CAJ | 학술논문

目的探讨幼年鼠急性肺损伤(acute lung injury,ALI)肺组织核转录因子(nuclear factor,NF)-κB及肿瘤坏死因子(tumor necrosis factor,TNF)-α的变化,观察ALI动物模型应用骨髓间充质干细胞(mesenchymal stem cells,MSCs)干预后肺组织损伤的修复程度.方法 30只雄性C57幼年鼠采用随机数字表法随机分为3组:模型组、对照组、模型± MSCs组(实验组),每组再分为12h、48h2个亚组(每组各5只).模型组腹腔注射脂多糖(lipopolysaccharide,LPS)(10 mg/kg)建立幼年鼠ALI模型,对照组同期注入等容积生理盐水,实验组在腹腔注射LPS同时经尾静脉注射MSCs(1×106/ml)0.1 ml.观察肺组织病理变化并应用免疫组织化学法检测NF-κB及TNF-α蛋白表达的变化.结果与对照组相比,模型组12h、48 h时,幼年鼠肺组织NF-κB、TNF-α蛋白表达显著增高(P＜0.05),而实验组12h、48 h时幼年鼠肺组织NF-κB、TNF-α蛋白表达较模型组显著降低[NF-κB:12 h:(0.181 ±0.008)OD值vs (0.203±0.008) OD值,48 h:(0.197±0.002) OD值vs (0.210±0.005) OD值;TNF-α:12 h:(0.185±0.004)OD值vs (0.201±0.011)OD值,48 h:(0.185 ±0.002) OD值vs (0.215±0.009)OD值](P＜0.05).病理学观察显示:模型组鼠肺组织呈现典型的炎症病理变化,包括肺泡充血、出血、水肿,肺泡腔和血管壁中性粒细胞浸润,肺泡壁增厚等肺损伤性病变;实验组肺组织损伤程度明显减轻.结论幼年鼠LPS致ALI模型组肺组织NF-κB及TNF-α蛋白表达增高,LPS致ALI的发病机制可能与NF-κB及TNF-α蛋白表达升高有关.应用MSCs干预后,实验组12h、48 h时幼年鼠肺组织NF-κB、TNF-α蛋白表达较模型组显著降低,实验组肺组织损伤减轻,提示MSCs能够减轻ALI肺组织的炎症反应并参与了肺组织损伤的修复.
목적 탐토유년서급성폐손상(acute lung injury,ALI)폐조직핵전록인자(nuclear factor,NF)-κB급종류배사인자(tumor necrosis factor,TNF)-α적변화,관찰ALI동물모형응용골수간충질간세포(mesenchymal stem cells,MSCs)간예후폐조직손상적수복정도.방법 30지웅성C57유년서채 용수궤수자표법수궤분위3조:모형조、대조조、모형± MSCs조(실험조),매조재분위12h、48h2개아조(매조각5지).모형조복강주사지다당(lipopolysaccharide,LPS)(10 mg/kg)건립유년서ALI모형,대조조동기주입등용적생리염수,실험조재복강주사LPS동시경미정맥주사MSCs(1×106/ml)0.1 ml.관찰폐조직병리변화병응용면역조직화학법검측NF-κB급TNF-α단백표체적변화.결과 여대조조상비,모형조12h、48 h시,유년서폐조직NF-κB、TNF-α단백표체현저증고(P＜0.05),이실험조12h、48 h시유년서폐조직NF-κB、TNF-α단백표체교모형조현저강저[NF-κB:12 h:(0.181 ±0.008)OD치vs (0.203±0.008) OD치,48 h:(0.197±0.002) OD치vs (0.210±0.005) OD치;TNF-α:12 h:(0.185±0.004)OD치vs (0.201±0.011)OD치,48 h:(0.185 ±0.002) OD치vs (0.215±0.009)OD치](P＜0.05).병이학관찰현시:모형조서폐조직정현전형적염증병리변화,포괄폐포충혈、출혈、수종,폐포강화혈관벽중성립세포침윤,폐포벽증후등폐손상성병변;실험조폐조직손상정도명현감경.결론 유년서LPS치ALI모형조폐조직NF-κB급TNF-α단백표체증고,LPS치ALI적발병궤제가능여NF-κB급TNF-α단백표체승고유관.응용MSCs간예후,실험조12h、48 h시유년서폐조직NF-κB、TNF-α단백표체교모형조현저강저,실험조폐조직손상감경,제시MSCs능구감경ALI폐조직적염증반응병삼여료폐조직손상적수복.
Objective To explore the changes in expression levels of nuclear factor(NF)-κB,tumor necrosis factor(TNF)-e in the lungs of juvenile mice with acute lung injury(ALI) induced by lipopolysaccharide(LPS).And observe the repair of lung damage after intervening with exogenous mesenchymal stem cells(MSCs).Methods Thirty male juvenile C57 mice were randomly divided into the control group,the ALI group,and the ALI + MSCs group by the random number table method.Mice from each group were euthanized at 12 h and 48 h.The ALI model of juvenile mice was established by intraperitoneal injection of LPS 10 mg/kg.MSCs from mice bone marrow were isolated,cultured and amplified in vitro,and the MSCs (1 × 106/ml) 0.1 ml were given to mice via caudal vein.MSCs marker were identificated by flow cytometry.Pathomorphological changes of mice lung were observed under light microscope after Hematoxylin-Eosin staining.The protein expression changes of NF-κB,TNF-α were observed using immunohistochemistry.Resu]ts Compared with the control group,the protein expression levels of NF-κB,TNF-α were significantly higher at 12 h and 48 h in the lungs of the ALI group(P ＜ 0.05).While those in ALI + MSCs group were markedly lower at these time points than the ALI group [NF-κB:12 h:(0.181 ± 0.008) OD vs (0.203 ±0.008) OD,48 h:(0.197 ± 0.002) OD vs (0.210 ± 0.005) OD; TNF-α:12 h:(0.185 ± 0.004) OD vs (0.201 ± 0.011) OD,48 h:(0.185 ± 0.002) OD vs (0.215 ± 0.009) OD] (P ＜ 0.05).Histopathological evalution showed that typical pathological inflammation lesions in the lung were observed in ALI group,including alveolar congestion,hemorrhage,edema,infiltration of neutrophils in the airspace or vessel wall,thickness of the alveolar wall;pathological changes were relieved obviously in ALI + MSCs group.Conclusion The expression of NF-κB and TNF-α are increased in lung tissues in the juvenile mice model of ALI induced by LPS.MSCs can alleviate injury degree of ALI induced by LPS in mice,the mechanism of action may correlate with decreasing NF-κB and TNF-α content in lung tissue.