氯气吸入致大鼠肺血管内皮细胞的线粒体损伤及红景天苷的干预作用

doi:10.3969/j.issn.1004-616x.2018.01.006

癌变·畸变·突变 ›› 2018, Vol. 30 ›› Issue (1): 31-36,41.doi: 10.3969/j.issn.1004-616x.2018.01.006

氯气吸入致大鼠肺血管内皮细胞的线粒体损伤及红景天苷的干预作用

刘萌萌, 李彦文, 周庆彪, 张佳欣, 王乐乐, 刘江正, 于卫华, 孔德钦, 刘瑞, 海春旭, 张晓迪

空军军医大学军事预防医学系毒理学教研室, 陕西西安 710032

收稿日期:2017-11-23 修回日期:2018-01-09 出版日期:2018-01-30 发布日期:2018-01-30
通讯作者: 张晓迪,E-mail:zhangxiaodi@fmmu.edu.cn E-mail:zhangxiaodi@fmmu.edu.cn
作者简介:刘萌萌,E-mail:liumeng1209@126.com。
基金资助:
军事医学创新工程专项（16CXZ021）；陕西省自然科学基金重点项目（2014J2009）

Effects of salidroside on mitochondria of lung in rats with chlorine-induced acute lung injury

LIU Mengmeng, LI Yanwen, ZHOU Qingbiao, ZHANG Jiaxin, WANG Lele, LIU Jiangzheng, YU Weihua, KONG Deqin, LIU Rui, HAI Chunxu, ZHANG Xiaodi

Department of Toxicology, School of Public Health, Medical University of the Air Force, Xi'an 710032, Shaanxi, China

Received:2017-11-23 Revised:2018-01-09 Online:2018-01-30 Published:2018-01-30

摘要/Abstract

摘要： 目的：观察氯气致大鼠肺损伤过程中，线粒体的结构与功能变化及红景天苷的干预作用，探讨红景天苷可能的保护机制。方法：36只雄性SD大鼠随机分为6组，分别为未染毒组（给予空气），以及氯气染毒后0.5、1.5、3、6和9 h组，给予1 200 mg/m³氯气暴露处理5 min。另24只雄性SD大鼠随机分为4组，分别为氯气暴露组、红景天苷干预组、红景天苷对照组和阴性对照组，氯气染毒条件同上，红景天苷干预组和红景天苷对照组在氯气染毒前30 min和染毒后15 min各给予1次300 mg/kg红景天苷灌胃，阴性对照组给予生理盐水灌胃。比色法检测各组大鼠肺泡灌洗液（BALF）和血清中乳酸脱氢酶（LDH）的活力；透射电镜下观察氯气暴露后3 h肺组织血管内皮细胞线粒体超微结构改变；定磷法检测肺组织Na⁺，K⁺-ATP酶和Ca²⁺，Mg²⁺-ATP酶的活力，低温差速离心法分离并提取线粒体蛋白；Western blot法检测肺组织或线粒体中Tom 20、PINK1以及Parkin蛋白表达水平。结果：与未染毒组相比，氯气暴露0.5 h后，大鼠BALF中LDH活力显著升高（P < 0.05）；肺组织PINK1、Parkin、Tom20蛋白表达水平均显著升高（P < 0.05），且Tom20蛋白表达在染毒后3 h最高（P < 0.05）；故选取染毒后3 h为干预时间点。透射电镜结果显示，氯气暴露后3 h，肺血管内皮细胞中线粒体明显肿胀，嵴数量减少或消失。与阴性对照组相比，氯气染毒后大鼠肺组织中PINK1、Parkin以及线粒体PINK1蛋白表达显著升高（P < 0.05），线粒体Parkin蛋白表达显著下降（P < 0.05）；肺组织Na⁺，K⁺-ATP酶和Ca²⁺，Mg²⁺-ATP酶以及血清LDH的活力均显著升高（P < 0.05）。红景天苷干预可显著逆转因氯气暴露引起的上述检测指标的异常表达（P < 0.05）。结论：氯气致大鼠急性肺损伤过程中，血管内皮细胞线粒体结构与功能受损，呈时间效应关系。红景天苷可能通过保护线粒体，改善氯气暴露引起的急性肺损伤。

关键词: 氯气, 线粒体, 急性肺损伤, 红景天苷

Abstract: OBJECTIVE: To observe the effects of chlorine on structure and function of mitochondria of lungs in rats and explore the protective effects of salidroside on acute lung injury induced by chlorine. METHODS: 36 healthy male Sprague-Dawley (SD) rats were divided into 6 groups:1 control and 5 exposure. Rats in the exposure groups were exposed to an initial concentration of 1 200 mg/m³ chlorine for 5 min in a devised cabinet or room air. Rats were sacrificed at 0.5,1.5,3,6 and 9 h,respectively,after exposure. In addition,24 rats were randomly divided into 4 groups:control,chlorine,chlorine+salidroside and salidroside. Chlorine and chlorine+salidroside groups were exposed to chlorine as mentioned above. Chlorine+salidroside and salidroside groups were treated with salidroside (300 mg/kg) at 30 min before chlorine exposure and at 15 min after chlorine exposure. Then the rats were sacrificed at 3 h after chlorine exposure,and the lung tissues were harvested. Lung mitochondria ultra structures were observed with electron microscope. The Na⁺,K⁺-ATPase,Ca²⁺,Mg²⁺-ATPase in lung tissues and lactate dehydrogenase (LDH) in serum and bronchoalveolar lavage fluid (BALF) were measured using kits,respectively. The mitochondria in lung tissues were isolated with differential centrifugation. The protein levels of Tom 20,PINK1 and Parkin in lung tissues or mitochondria were detected by western blotting. RESULTS: Compared with control group,LDH of BALF in exposure groups were significantly increased (P < 0.05). Tom 20,PINK1 and Parkin protein expressions in lung tissues were all higher than that in control group (P < 0.05). Expression of Tom 20 was highest at 3 h. It was also shown by transmission electron microscope that the mitochondria structure in the control group was normal. The mitochondria in rat pulmonary vascular endothelial cells were swollen with disrupted or disintergrated cristae. When treated with salidroside,the expression levels of PINK1 and Parkin in lung tissues and PINK1 in mitochondria were reduced compared with chlorine group at 3 h. The expressions of Parkin in mitochondria was markedly increased (P < 0.05);Na⁺,K⁺-ATPase,Ca²⁺,Mg²⁺-ATPase in lung tissues and LDH in serum were decreased. CONCLUSION: Chlorine induced mitochondrial structural damage and functional impairment in rats and salidroside exerted a beneficial effect through protecting the mitochondria.

Key words: chlorine, mitochondria, acute lung injury, salidroside

中图分类号:

R961

刘萌萌, 李彦文, 周庆彪, 张佳欣, 王乐乐, 刘江正, 于卫华, 孔德钦, 刘瑞, 海春旭, 张晓迪. 氯气吸入致大鼠肺血管内皮细胞的线粒体损伤及红景天苷的干预作用[J]. 癌变·畸变·突变, 2018, 30(1): 31-36,41.

LIU Mengmeng, LI Yanwen, ZHOU Qingbiao, ZHANG Jiaxin, WANG Lele, LIU Jiangzheng, YU Weihua, KONG Deqin, LIU Rui, HAI Chunxu, ZHANG Xiaodi. Effects of salidroside on mitochondria of lung in rats with chlorine-induced acute lung injury[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2018, 30(1): 31-36,41.

参考文献

[1] Van SICKLE D,WENCK M A,BELFLOWER A,et al. Acute health effects after exposure to chlorine gas released after a train derailment[J]. Am J Emerg Med,2009,27(1):1-7.
[2] JURKUVENAITE A,BENAVIDES G A,KOMAROVA S,et al. Upregulation of autophagy decreases chlorine gas induced mitochondrial injury and lung inflammation[J]. Free Radic Biol Med,2015,85:83-94.
[3] DEAS E,PLUN-FAVREAU H,WOOD N W. PINK1 function in health and disease[J]. Embo Mol Med,2009,1(3):152-165.
[4] SCARFFE L A,STEVENS D A,DAWSON V L,et al. Parkin and PINK1:much more than mitophagy[J]. Trends Neurosci,2014,37(6):315-324.
[5] 张晓迪,赵琰,李文丽,等. 红景天苷对氯气致大鼠肺损伤的保护作用[J]. 预防医学情报杂志,2013,29(4):269-272.
[6] 尚涛,李立萍,张建新. 线粒体与急性肺损伤[J]. 河北医药,2007,29(4):367-369.
[7] ISLAM M N,DAS S R,EMIN M T,et al. Mitochondrial transfer from bone-marrow-derived stromal cells to pulmonary alveoli protects against acute lung injury[J]. Nat Med,2012,18(5):759-765.
[8] 焦光宇,毕涛,高鸿美,等. 内毒素对大鼠膈肌收缩功能及线粒体超微结构的影响[J]. 中华结核和呼吸杂志,2008,31(6):431-435.
[9] 刘萌萌,孔德钦,邹远康,等. 红景天苷对氯气暴露致急性肺损伤肺血管通透性的保护作用[J]. 癌变·畸变·突变,2016,28(5):377-382.
[10] ZHONG H,XIN H,WU L X,et al. Salidroside attenuates apoptosis in ischemic cardiomyocytes:a mechanism through a mitochondria-dependent pathway[J]. J Pharmacol Sci,2010,114(4):399-408.
[11] 章娟. 大花红景天质量控制和相关成分药代动力学研究[D]. 沈阳:沈阳药科大学,2008.
[12] VALENTE E M,ABOU-SLEIMAN P M,CAPUTO V,et al. Hereditary early-onset Parkinson's disease caused by mutations in PINK1[J]. Science,2004,304(5674):1158-1160.
[13] BEILINA A,Van Der BRUG M,AHMAD R,et al. Mutations in PTEN-induced putative kinase 1 associated with recessive parkinsonism have differential effects on protein stability[J]. Proc Natl Acad Sci U S A,2005,102(16):5703-5708.
[14] KITADA T,ASAKAWA S,HATTORI N,et al. Mutations in the parkin gene cause autosomal recessive juvenile parkinsonism[J]. Nature,1998,392(6676):605-608.
[15] GEGG M E,COOPER J M,SCHAPIRA A H,et al. Silencing of PINK1 expression affects mitochondrial DNA and oxidative phosphorylation in dopaminergic cells[J]. PLoS One,2009,4(3):e4756.
[16] MORAIS V A,VERSTREKEN P,ROETHIG A,et al. Parkinson's disease mutations in PINK1 result in decreased complex I activity and deficient synaptic function[J]. Embo Mol Med,2009,1(2):99-111.
[17] ROTHFUSS O,FISCHER H,HASEGAWA T,et al. Parkin protects mitochondrial genome integrity and supports mitochondrial DNA repair[J]. Hum Mol Genet,2009,18(20):3832-3850.
[18] ZHANG H,MI L,WANG T,et al. PINK1/Parkin-mediated mitophagy play a protective role in manganese induced apoptosis in SH-SY5Y cells[J]. Toxicol in Vitro,2016,34:212-219.
[19] NARENDRA D P,JIN S M,TANAKA A,et al. PINK1 is selectively stabilized on impaired mitochondria to activate Parkin[J]. PLoS Biol,2010,8(1):e1000298.
[20] 张伟,李涛,陈磊,等. 红景天苷通过PINK1-Parkin通路在MPP+诱导的SH-SY5Y细胞中维持线粒体形态和功能[J]. 现代生物医学进展,2016,16(9):1649-1653.
[21] QIN X,LI Y,LIANG X,et al. The dysfunction of ATPases due to impaired mitochondrial respiration in phosgene-induced pulmonary edema[J]. Biochem Bioph Res Co,2008,367(1):150-155.
[22] MCMAHON S,CHARBONNEAU M,GRANDMONT S,et al. Transforming growth factor beta1 induces hypoxia-inducible factor-1 stabilization through selective inhibition of PHD2 expression[J]. J Biol Chem,2006,281(34):24171-24181.

氯气吸入致大鼠肺血管内皮细胞的线粒体损伤及红景天苷的干预作用

Effects of salidroside on mitochondria of lung in rats with chlorine-induced acute lung injury

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	尹亚萍, 王宇, 田雨阳, 马焜, 李百祥. 褪黑素在帕金森病治疗中的研究进展[J]. 癌变·畸变·突变, 2022, 34(1): 75-78,81.
[2]	兰尤, 陈圆圆, 林忠宁, 林育纯. 病毒致癌过程中的线粒体糖酵解代谢重编程机制研究进展[J]. 癌变·畸变·突变, 2021, 33(6): 470-474.
[3]	陈廷玉, 陈大印, 谢金鹿, 高喜仁, 卢春凤. Nrf2/ARE信号通路在槲皮素抑制异烟肼诱导的肝细胞线粒体氧化损伤中的作用[J]. 癌变·畸变·突变, 2021, 33(3): 208-212,217.
[4]	陈廷玉, 陈大印, 沈洪, 富徐燕, 卢春凤. 槲皮素对异烟肼诱导肝细胞毒性的保护作用及其机制[J]. 癌变·畸变·突变, 2021, 33(1): 12-16.
[5]	师盼, 林重华, 舒易方, 瞿家权, 宋淑亮. 柠檬酸转运体SLC25A1对食管鳞癌细胞体外放射敏感性的影响及其分子机制[J]. 癌变·畸变·突变, 2020, 32(2): 132-138.
[6]	郭倪君, 林忠宁, 林育纯. 铁死亡信号介导抗肿瘤作用的细胞器调控机制研究进展[J]. 癌变·畸变·突变, 2020, 32(2): 149-154.
[7]	张虹, 王彤, 王坤, 王博深, 章梦莹, 周燕华, 浦跃朴, 张娟. Camk2b低表达对1,4-苯醌致K562细胞线粒体毒性的影响[J]. 癌变·畸变·突变, 2019, 31(4): 261-267.
[8]	梁辰, 靳翠红. 乳腺癌易感基因1调控氧化损伤在神经退行性疾病中作用的研究进展[J]. 癌变·畸变·突变, 2019, 31(3): 257-260.
[9]	车琳, 林忠宁, 林育纯. 蛋白磷酸酶2A在线粒体质量控制中的作用研究进展[J]. 癌变·畸变·突变, 2019, 31(1): 73-78.
[10]	陈文龙, 由淑萍, 赵军, 张杰, 刘涛. 阿尔泰金莲花浸膏粉对PM2.5致大鼠急性肺损伤的保护作用[J]. 癌变·畸变·突变, 2019, 31(1): 22-28.
[11]	郑义鹏, 魏学敏, 周庆彪, 赵九洲, 张晓迪, 孔德钦, 海春旭, 于卫华. 线粒体分裂融合与细胞氧化还原交互调控作用的研究进展[J]. 癌变·畸变·突变, 2018, 30(3): 239-241,247.
[12]	姚欢, 林育纯, 林忠宁. 线粒体关联性内质网膜与细胞Ca²⁺依赖性死亡的研究进展[J]. 癌变·畸变·突变, 2018, 30(3): 242-247.
[13]	张佳欣, 刘颖, 张伟, 周庆彪, 孔德钦, 刘瑞, 海春旭. TGF-β1诱导上皮间质转化过程中线粒体合成与功能的改变[J]. 癌变·畸变·突变, 2017, 29(6): 411-417.
[14]	韩孝先, 刘泽宇, 周庆彪, 张晓迪, 孔德钦, 海春旭, 于卫华. 线粒体在炎症调控中的作用研究进展[J]. 癌变·畸变·突变, 2017, 29(6): 467-470,475.
[15]	蒋小云, 孙蓉丽, 满招娣, 张娟, 浦跃朴. 1,4-苯醌致K562细胞线粒体功能障碍与凋亡作用[J]. 癌变·畸变·突变, 2017, 29(2): 129-133.