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寒冷刺激支气管上皮细胞产生外泌体促肺成纤维细胞表达气道重塑相关因子 被引量:1

Exosomes Produced by Cold-stimulated Bronchial Epithelial Cells Promote Expression of Airway Remodeling-related Factors in Lung Fibroblasts
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摘要 外泌体可由多种细胞分泌,是具有多种生物学功能的细胞外囊泡,但其在气道重塑中的作用尚不明确。为探讨经寒冷刺激的人支气管上皮细胞(BEAS-2B)分泌的外泌体对人胚肺成纤维细胞(HLF1)气道重塑相关因子表达的影响,收集BEAS-2B细胞株培养液提取外泌体,利用透射电镜及Western印迹对外泌体进行其大小、形态及标志性蛋白的检测;提取的外泌体与HLF1共同培育,分别设置空白对照组、正常对照组(加入未作干预的BEAS-2B细胞所产的外泌体)及寒冷刺激组(加入经寒冷刺激后的BEAS-2B细胞所产的外泌体)。运用Real-time PCR及Western印迹技术,分别检测各组HLF1表达FGF-2、TNF-α、MMP-9的mRNA及蛋白情况。结果显示,提取BEAS-2B细胞分泌的外泌体为直径小于100 nm的圆形或椭圆形结构,并表达外泌体标志性蛋白CD9、TSG101、ALIX;寒冷刺激组24 h后,其FGF-2、TNF-α、MMP-9的mRNA及蛋白表达均显著高于空白对照组及正常对照组(均P〈0.05)。本研究结果表明,BEAS-2B细胞能够释放外泌体;经寒冷刺激后的BEAS-2B细胞所释放的外泌体可以携带并传递生物信号,诱导HLF1表达气道重塑相关因子。 Exosomes are extracellular vesicles secreted by various cells. They have multiple biological functions, but their roles in airway remodeling remain uncertain. The study aims to explore the effects of cold-stimulated exosomes secreted by human bronchial epithelial cells (BEAS-2B) on the expression of airway remodeling-related factors in human embryonic lung fibroblasts (HLF1). The exosomes were identified by transmission electron microscope (TEM) and Western blotting analysis after extracted from BEAS-2B cell supernatants. The extracted exosomes were then incubated with HLF1 cells. The blank control group (conventional culture) , the normal control group (exosomes secreted by BEAS-2B cells without intervention) and the cold stimulation group (exosomes produced by BEAS-2B cells incubated in cold temperatures) were used. The results showed that the exosomes secreted by BEAS-2B cells wereround or oval with diameters less than 100 nm, and expressed the exosome markers cDg, TSG101 and ALIX. By real-time PCR and Western blotting, we found that the mRNA and protein levels of fibroblast growth factors-2 (FGF-2), tumor necrosis factor-or (TNF-a) and matrix metalloprotein-9 (MMP-9) in the cold stimulation group were significantly higher than those in the blank and normal control groups (both P 〈 0.05). The results showed that BEAS-2B cells could release exosomes. The exosomes released by BEAS-2B cells after cold stimulation could carry and transmit biological signals and induce HLF1 cells to produce airway remodeling-related factors.
作者 朱芮 李敏超 ZHU Rui, LI Min-Chao ( Department of Respiratory Medicine, The Second Affiliated Hospital of Chongqing Medical University Chongqing400010, China)
出处 《中国生物化学与分子生物学报》 CSCD 北大核心 2017年第10期1070-1075,共6页 Chinese Journal of Biochemistry and Molecular Biology
基金 国家自然科学基金面上基金(N0.81270102) 重庆市自然科学基金(No.cstc2012jjA10050) 重庆市教委科学技术研究(No.KJ120301)资助
关键词 寒冷刺激 外泌体 气道重塑 人支气管上皮细胞 人胚肺成纤维细胞 cold stimulation exosomes airway remodeling human bronchial epithelial cells humanembryonic lung fibroblast cells
作者简介 通讯作者Tel:023-63693758;E—mail:lirainc66@aliyun.Com
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