Protective role of mesenchymal stem cells and mesenchymal stem cell-derived exosomes in cigarette smoke-induced mitochondrial dysfunction in mice
Protective role of mesenchymal stem cells and mesenchymal stem cell-derived exosomes in cigarette smoke-induced mitochondrial dysfunction in mice
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DOI：
10.1016/j.taap.2019.114788
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发表时间：
2019-12-15
影响因子：
3.8
通讯作者：
Rahman, Irfan
中科院分区：
文献类型：
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作者：
Maremanda, Krishna Prahlad;Sundar, Isaac Kirubakaran;Rahman, Irfan
背景：香烟烟雾（CS）诱导的肺部炎症和慢性阻塞性肺疾病（COPD）涉及线粒体功能障碍。据报道，间充质干细胞（MSC）和间充质干细胞衍生的外泌体（EXO）在许多炎症和损伤动物模型中显示出治疗作用。在本研究中，我们确定了MSC和EXO干预在cs诱导的肺部炎症中的作用，重点是线粒体功能障碍。方法：采用Western blot检测外泌体标记，qNano可调电阻脉冲传感和透射电子显微镜（TEM）对EXO进行表征。线粒体报告小鼠（mt-Keima和mitto - qc）暴露于空气或CS中10天。给mt-Keima小鼠腹腔注射MSC或EXO或MSC和EXO （MSC + EXO）治疗10天。用自动细胞计数仪和流式细胞仪分别进行总细胞计数、差异细胞计数。此外，采用ELISA法测定支气管肺泡灌洗液（BAL）中促炎介质的水平。本研究采用Western blot分析、定量PCR、共聚焦显微镜检测CS对小鼠肺部的影响。采用海马通量分析仪测定BEAS2B细胞氧化磷酸化（OXPHOS）及BEAS2B - mMSC共培养实验。结果：CS暴露增加了mt-Keima小鼠肺部炎症细胞浸润。与单独治疗（MSC或EXO）相比，MSC + EXO治疗显示出保护作用。在使用mito-QC小鼠时也发现，有丝分裂蛋白如PINK1和Parkin没有变化。CS暴露导致线粒体裂变蛋白DRP1和其他DAMPs通路介质如S100A4和S100A8、HMGB1、RAGE和AGE显著增加。MSC + EXO处理增加了（融合基因）mfn1、mfn2和opa1的基因表达。此外，与空气和CS暴露组相比，MSC + EXO处理组rhot1基因表达增加。BEAS2B-mMSC共培养对cse改变的线粒体呼吸参数表现出保护反应，证实了MSC对人支气管肺上皮细胞的有益作用。结论：CS影响了参与裂变/融合过程的一些早期线粒体基因，增强了损伤反应，并改变了细胞因子水平。MSC + EXO联合治疗显示出保护作用。MSC + EXO联合治疗可能由于其抗炎和其他线粒体转移机制而对CS暴露引起的这些早期事件起作用。
Background: Cigarette smoke (CS)-induced lung inflammation and Chronic Obstructive Pulmonary disease (COPD) involves mitochondrial dysfunction. Mesenchymal stem cells (MSC) and MSC-derived exosomes (EXO) are reported to show therapeutic effects in many animal models of inflammation and injury. In the present study, we determined the role of MSC and EXO intervention in CS-induced lung inflammation with a focus on mitochondrial dysfunction.Methods: EXO were characterized using Western blot for exosomal markers, tunable resistive pulse sensing by qNano and transmission electron microscopy (TEM). Mitochondrial reporter mice (mt-Keima and mito-QC) were exposed to air or CS for 10 days. mt-Keima mice were treated with intraperitoneal injections of MSC or EXO or MSC and EXO (MSC + EXO) for 10 days. Total cell counts, differential cell counts were performed using automated cell counter and flow cytometry respectively. Further, the levels of pro-inflammatory mediators in bronchoalveolar lavage (BAL) fluid were measured using ELISA. Western blot analysis, quantitative PCR, confocal microscopy were used in the current study to determine the effects in the lungs of CS exposed mice. Seahorse flux analyzer was used to measure the oxidative-phosphorylation (OXPHOS) in the BEAS2B cells and BEAS2B - mMSC co-culture experiments.Results: CS exposure increased the inflammatory cellular infiltrations in the lungs of the mt-Keima mice. MSC + EXO treatment showed protection compared to individual treatments (MSC or EXO alone). There were no changes in the mitophagy proteins like PINK1 and Parkin, which was also found using the mito-QC mice. CS exposure led to significant increase in the mitochondrial fission protein DRP1 and other DAMPs pathway mediators like S100A4 and S100A8, HMGB1, RAGE and AGE. MSC + EXO treatment increased the gene expression of (fusion genes) mfn1, mfn2 and opa1. Additionally, the rhot1 gene expression was increased in MSC + EXO treatment group compared to Air- and CS exposed groups. BEAS2B-mMSC co-cultures showed protective response against the CSE-altered mitochondrial respiration parameters, confirming the beneficial effect of MSC towards human bronchial lung epithelial cells.Conclusion: CS affects some of early mitochondrial genes involved in the fission/fusion process, enhancing the damage response along with altered cytokine levels. MSC + EXO combination treatment showed their protective effects. MSC + EXO combination treatment may act against these early events caused by CS exposure owing to its anti-inflammatory and other mitochondrial transfer mechanisms.