中国医科大学学报

中国医科大学学报
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中国医科大学学报 ›› 2019, Vol. 48 ›› Issue (1): 1-6.doi: 10.12007/j.issn.0258-4646.2019.01.001

• 论著 •    下一篇

MnTMPyP通过抑制氧化应激和内质网应激减轻百草枯致肺上皮细胞损伤

许勇民, 孙大壮, 宋春青, 王蕊, 董雪松   

  1. 中国医科大学附属第一医院急诊科, 沈阳 110001
  • 收稿日期:2018-03-27 出版日期:2019-01-30 发布日期:2019-01-05
  • 通讯作者: 董雪松 E-mail:dongxues@163.com
  • 作者简介:许勇民(1987-),男,硕士研究生.
  • 基金资助:
    国家自然科学基金(81471851);辽宁省博士启动基金(20141033)

MnTMPyP Reduces Paraquat-induced Lung Epithelial-like Cell Injury by Inhibiting Oxidative Stress and Endoplasmic Reticulum Stress

XU Yongmin, SUN Dazhuang, SONG Chunqing, WANG Rui, DONG Xuesong   

  1. Department of Emergency, The First Hospital, China Medical University, Shenyang 110001, China
  • Received:2018-03-27 Online:2019-01-30 Published:2019-01-05

摘要: 目的 探讨锰(Ⅲ)-四(4-N-甲基吡啶基)卟啉(MnTMPyP)对百草枯(PQ)致肺上皮细胞损伤是否具有保护作用及相关机制。方法 体外培养人肺泡Ⅱ型上皮样细胞A549细胞,以超氧化物歧化酶的模拟物MnTMPyP为干预因素。实验分为4组:对照组,加入RPMI 1640培养液;MnTMPyP组,加入MnTMPyP 10 μmol/L;PQ组,加入PQ 750 μmol/L;PQ+MnTMPyP组,MnTMPyP 10 μmol/L预处理90 min,再加入PQ 750 μmol/L。各组细胞处理24 h后检测相关指标。MTT法检测细胞活性;流式细胞术检测活性氧(ROS)、细胞质内Ca2+水平;紫外比色法检测谷胱甘肽还原酶活性;Western blotting检测内质网应激蛋白葡萄糖调节蛋白78(Grp78)、C/EBP同源蛋白(CHOP)蛋白的表达。结果 与对照组相比,MnTMPyP组检测指标的变化无统计学意义;PQ组细胞活性降低,ROS产生显著增加,细胞质内Ca2+水平明显升高,谷胱甘肽还原酶活性明显降低,Western blotting显示内质网应激蛋白Grp78、CHOP表达量均显著增加。与PQ组相比,PQ+MnTMPyP组细胞活性增加,ROS产生减少,细胞质内Ca2+水平降低,谷胱甘肽还原酶活性增加,Grp78、CHOP蛋白表达量均显著减少。结论 MnTMPyP有效减轻PQ诱导的肺上皮细胞损伤,其机制与拮抗PQ引起的氧化应激和内质网应激有关。

关键词: MnTMPyP, 百草枯, 肺上皮细胞, 氧化应激, 内质网应激

Abstract: Objective To investigate the protective effect and underlying mechanism of the superoxide dismutase mimic,manganese (Ⅲ) tetrakis (1-methyl-4-pyridyl) porphyrin pentachloride (MnTMPyP),on paraquat (PQ) -induced lung epithelial-like cell injury. Methods alveolar epithelial-like cells (A549) were pretreated with 10 μmol/L of MnTMPyP for 1.5 h and then cultured with or without PQ (750 μmol/L) for 24 h. Cell survival was determined using the MTT assay. Reactive oxygen species (ROS) production and Ca2+ levels were measured using flow cytometry. Glutathione reductase (GR) activity was determined using spectrophotometry. Expressions of the endoplasmic reticulum (ER) stress proteins,glucose regulatory protein 78 (Grp78) and C/EBP homologous protein (CHOP),were measured using Western blotting. Results Cell viability and GR activity were decreased,but ROS production,cytoplasmic Ca2+ levels,and expressions of Grp78 and CHOP were all increased in the PQ group compared to those in the control group. There were no statistically significant changes in the MnTMPyP group. Cell viability and GR activity were increased,while ROS production,cytoplasmic Ca2+ levels,and expressions of Grp78 and CHOP were all significantly reduced in the MnTMPyP group compared to those in the PQ group. Conclusion MnTMPyP effectively reduced PQ-induced lung epithelial-like cell injury,and the underlying mechanism is related to antagonism of PQ-induced ER stress and oxidative stress.

Key words: MnTMPyP, paraquat, lung epithelial-like cell, oxidative stress, endoplasmic reticulum stress

中图分类号: 

  • R595.4
[1] ZHANG BL,YAO L,OU Y. Literature analysis of paraquat poisoning in China (1991-2008)[J]. Chin J Crit Care Med,2010,30(2):139-141. DOI:10.3969/j.issn.1002-1949.2010.02.013.
[2] DINIS-OLIVEIRA RJ,DUARTE JA,S NCHEZ-NAVARRO A,et al. Paraquat poisonings:mechanisms of lung toxicity,clinical features and treatment[J]. Crit Rev Toxicol,2008,38(1):13-71.
[3] BLANCO-AYALA T,ANDERICA-ROMERO AC,PEDRAZA-CHAV-ERRI J. New insights into antioxidant strategies against paraquat toxicity[J]. Free Radic Res,2014,48(6):623-640. DOI:10.3109/10715762.2014.899694.
[4] SHOKRZADEH M,SHAKI F,MOHAMMADI E,et al. Edaravone decreases paraquat toxicity in a549 cells and lung isolated mitochondria[J]. Iran J Pharm Res,2014,13(2):675-681.
[5] POURGHOLAMHOSSEIN F,RASOOLI R,POURNAMDARI M,et al. Pirfenidone protects against paraquat-induced lung injury and fibrosis in mice by modulation of inflammation,oxidative stress,and gene expression[J]. Food Chem Toxicol,2018,112:39-46. DOI:10.1016/j.fct.2017.12.034.
[6] JAVAD-MOUSAVI SA,HEMMATI AA,MEHRZADI S,et al. Protective effect of Berberis vulgaris fruit extract against Paraquat-induced pulmonary fibrosis in rats[J]. Biomed Pharmacother,2016,81:329-336. DOI:10.1016/j.biopha.2016.04.027.
[7] SHEN HN,WU Y,WANG H,et al. Chloroquine attenuates paraquat-induced lung injury in mice by altering inflammation,oxidative stress and fibrosis[J]. Int Immunopharmacol,2017,46:16-22. DOI:10.1016/j.intimp.2017.02.020.
[8] COCHEME HM,MURPHY MP. Complex Ⅰ is the major site of mitochondrial superoxide production by paraquat[J]. Biol Chem,2008,283(4):1786-1798. DOI:10.1074/jbc. M708597200.
[9] PATKI G,LAU YS. Melatonin protects against neurobehavioral and mitochondrial deficits in a chronic mouse model of Parkinson's disease[J]. Pharmacol Biochem Behav,2011,99(4):704-711. DOI:10.1016/j.pbb.2011.06.026.
[10] KIRBY K,HU J,HILLIKER AJ,et al. RNA interference-mediated silencing of Sod2 in Drosophila leads to early adult-onset mortality and elevated endogenous oxidative stress[J]. Proc Natl Acad Sci U S A,2002,99(25):16162-16167. DOI:10.1073/pnas.252342899.
[11] DIXIT A,SRIVASTAVA G,VERMA D. Minocycline,levodopa and MnTMPyP induced changes in the mitochondrial proteome profile of MPTP and maneb and paraquat mice models of Parkinson's disease[J]. Biochim Biophys Acta,2013,1832(8):1227-1240. DOI:10.1016/j.bbadis.2013.03.019.
[12] HAN SJ,JANG HS,SEU SY,et al. Hepatic ischemia/reperfusion injury disrupts the homeostasis of kidney primary cilia via oxidative stress[J]. Biochim Biophys Acta,2017,1863(7):1817-1828. DOI:10.1016/j.bbadis.2017.05.004.
[13] KIM JI,KIM J,JANG HS,et al. Reduction of oxidative stress during recovery accelerates normalization of primary cilia length that is altered after ischemic injury in murine kidneys[J]. Am J Physiol Renal Physiol,2013,304(10):F1283-F1294. DOI:10.1152/ajprenal.00427.2012.
[14] CHOI IY,LEE SJ,JU C,et al. Protection by a manganese porphyrin of endogenous peroxynitrite-induced death of glial cells via inhibition of mitochondrial transmembrane potential decrease[J]. Glia,2000,31(2):155-164.
[15] SUNTRES ZE. Role of antioxidants in paraquat toxicity[J]. Toxicology,2002,180(1):65-77.
[16] LI X,WANG Y,WANG H,et al. Endoplasmic reticulum stress is the crossroads of autophagy,inflammation,and apoptosis signaling pathways and participates in liver fibrosis[J]. Inflamm Res,2015,64(1):1-7. DOI:10.1007/s00011-014-0772-y.
[17] PRELL T,LAUTENSCHLAGER J,GROSSKREUTZ J. Calcium-dependent protein folding in amyotrophic lateral sclerosis[J]. Cell Calcium,2013,54(2):132-143. DOI:10.1016/j.ceca.2013.05.007.
[18] CLARKE HJ,CHAMBERS JE,LINIKER E,et al. Endoplasmic reticulum stress in malignancy[J]. Cancer Cell,2014,25(5):563-573. DOI:10.1016/j.ccr.2014.03.015.
[19] ZHANG J,LI Y,JIANG S,et al. Enhanced endoplasmic reticulum SERCA activity by overexpression of hepatic stimulator substance gene prevents hepatic cells from ER stress-induced apoptosis[J]. Am J Physiol Cell Physiol,2014,306(3):C279-C290. DOI:10.1152/ajpcell.00117.2013.
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