脂氧素A_4对高氧损伤肺泡Ⅱ型细胞系MLE-12的保护作用  

Protective effect of lipoxin A_4 on hyperoxia injury of typeⅡalveolar epithelial cell line MLE-12

作　　者：李淑君[1] 李冰洁[1] 罗妍妍[1] 卢红艳[2] 吴升华[1] 陈筱青[1] 

机构地区：[1]南京医科大学第一附属医院儿科,江苏南京210029 [2]江苏大学附属医院儿科,江苏镇江212000

出　　处：《南京医科大学学报（自然科学版）》2018年第2期185-190,共6页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家自然科学基金资助(81300521;81270821;81370746)

摘　　要：目的:探讨脂氧素A_4(Lipoxin A_4,LXA_4)对高氧损伤肺泡Ⅱ型细胞系MLE-12的保护作用及机制。方法:体外传代培养MLE-12细胞,随机分为:空气组、高氧组、高氧+LXA_4组、高氧+转化生长因了(TGF)-β1中和抗体组、高氧+LXA_4+TGF-β1中和抗体组。倒置相差显微镜下观察各组MLE-12形态学变化;实时荧光定量PCR(q RT-PCR)法检测Ⅰ型胶原(collagenⅠ)、肌腱蛋白C(tenascin-C)、TGF-βR1、TGF-βR2和Smad3的m RNA水平。Western blot检测TGF-β1/Smads信号(TGF-βR1、Smad2、Smad3、Smad4、p-Smad2和p-Smad3)蛋白表达。结果:(1)细胞形态:高氧组MLE-12明显失去原有正常细胞形态,细胞变圆,核固缩;药物干预组正常细胞数增多,大部分细胞形态与正常细胞形态基本相似;(2)细胞外基质(extracellular matrix,ECM)m RNA表达:LXA_4和TGF-β1中和抗体能抑制高氧介导的collagenⅠ和tenascin-C m RNA的表达量升高(P<0.05),其中高氧+LXA_4+TGF-β1中和抗体组作用最为显著(P<0.05);(3)TGF-β1/Smads信号m RNA和蛋白含量:LXA_4和TGF-β1中和抗体能抑制高氧介导的TGF-βR1、TGF-βR2和Smad3的m RNA水平以及下调TGF-β1/Smads信号相关蛋白的表达量(P<0.05),其中高氧+LXA_4+TGF-β1中和抗体组细胞的表达量下调最为显著(P<0.05)。结论:LXA_4对高氧损伤肺泡Ⅱ型细胞系MLE-12的保护可能与调控TGF-β1/Smads信号以及collagenⅠ和tenascin-C的表达有关。Objective：To investigate protectory effects and possible mechanisms of lipoxin A4（LXA4）on the type Ⅱ alveolar epithelial cell line MLE-12 with hyperoxia injury. Methods：MLE-12 cells were cultured and divided into five groups randomly：I：air group,Ⅱ：hyperoxia group,Ⅲ：hyperoxia＋ LXA4 group,Ⅳ：hyperoxia＋ TGF-β1 neutralizing antibodies,Ⅴ：hyperoxia＋ TGF-β1 neutralizing antibodies＋ LXA4. The growth state and morphological changes of the cells were observed by the inverted phase microscope;the mRNA expression of collagen Ⅰ,tenascin-C,TGF-β1 receptor（TGF-βR1）,TGF-βR2 and Smad3 were measured by real-time quantitative PCR;the protein levels of TGF-β1/Smads signaling pathway（TGFβR1,Smad2,Smad3,Smad4,p-Smad2 and p-Smad3）were detected by Western blot. Results：（1）Morphological change：the cells under hyperoxia lost their normal shapes and showed necrosis,while the cells cultured with LXA4 and/lor TGF-β1 neutralizing antibodies were normal appropriately.（2）The mRNA expression of extracellular matrix（ECM）：LXA4 and TGF-β1 neutralizing antibodies attenuated the mRNA expression of collagenⅠ,tenascin-C markedly（P〈0.05）,which expressed in the group hyperoxia＋TGF-β1 neutralizing antibodies ＋ LXA4 decreased most obviously.（3）The mRNA and protein levels of TGF-β1/Smads signaling pathway：Compared with that of the hyperoxia group,the mRNA expression of TGF-βR1,TGF-βR2 and Smad3 decreased（P〈0.05）,while the protein level of TGF-β1/Smads signaling pathway had the same change in intervention group,and it in the the group hyperoxia＋ TGF-β1 neutralizing antibodies ＋LXA4 decreased most obviously（P〈0.05）. Conclusion：LXA4 can achieve protection of hy-peroxia-induced injury to MLE-12 which may be related to collagenⅠ,tenascin-C and TGF-β1/Smads signaling pathway.

关 键 词：脂氧素A_4 TGF-β1/Smads信号通路 细胞外基质 高氧损伤 

分 类 号：R329.26[医药卫生—人体解剖和组织胚胎学]