Email Alert | RSS

北京大学学报（医学版） ›› 2019, Vol. 51 ›› Issue (2): 239-244. doi: 10.19723/j.issn.1671-167X.2019.02.007

• 论著 • 上一篇    下一篇

二氧化硫对大鼠肢体缺血再灌注致急性肺损伤中肺泡巨噬细胞凋亡的影响

赵彦瑞,刘洋,王东,吕文睿,周君琳()   

  1. 首都医科大学附属北京朝阳医院骨科, 北京 100020
  • 收稿日期:2017-04-06 出版日期:2019-04-18 发布日期:2019-04-26
  • 通讯作者: 周君琳 E-mail:doctorzyr@163.com
  • 基金资助:
    国家自然科学基金(81070050);北京市自然科学基金(7152061)

Effects of sulfur dioxide on alveolar macrophage apoptosis in acute lung injury induced by limb ischemia/reperfusion in rats

Yan-rui ZHAO,Yang LIU,Dong WANG,Wen-rui LV,Jun-lin ZHOU()   

  1. Department of Orthopedics, Beijing Chao-yang Hospital, Capital Medical University, Beijing 100020, China
  • Received:2017-04-06 Online:2019-04-18 Published:2019-04-26
  • Contact: Jun-lin ZHOU E-mail:doctorzyr@163.com
  • Supported by:
    the National Natural Science Foundation of China(81070050);Beijing Natural Science Foundation(7152061)

RICH HTML

 13   

PDF (PC)

261

摘要：

目的: 探讨二氧化硫(sulfur dioxide,SO2)在肢体缺血再灌注(ischemia/reperfusion,I/R)致急性肺损伤(acute lung injury,ALI)保护作用中对肺泡巨噬细胞(alveolar macrophage,AM)凋亡的影响,为控制炎症反应寻找新的靶点。方法: 分离培养AM,应用肢体缺血再灌注致ALI大鼠血清制备细胞模型,给予外源性SO2,然后检测线粒体膜电位以及线粒体通透性转换孔(mitochondrial permeability transition pore,mPTP)开放情况,AM凋亡情况及凋亡相关Bcl-2、Caspase-3分子蛋白表达情况。结果: 与对照组相比,I/R组红、绿荧光的比值下降,吸光度显著降低,AM凋亡率增加到43.81%±2.40%,Caspase-3蛋白表达升高,Bcl-2蛋白表达下降;而与I/R组比较,I/R+SO2组红、绿荧光的比值升高,吸光度增高,AM凋亡率减少37.01%±1.93%,Caspase-3蛋白表达降低,Bcl-2蛋白表达升高。结论: 外源性SO2可通过抑制线粒体途径改善巨噬细胞的凋亡。

关键词: 二氧化硫, 再灌注损伤, 急性肺损伤, 巨噬细胞, 肺泡

Abstract:

Objective: To investigate the effect of sulfur dioxide (SO2) on the apoptosis of alveolar macrophage (AM) in lung protection of limb ischemia/reperfusion (I/R) induced acute lung injury (ALI), and to find a new target for the control of inflammatory response.Methods: Twenty pathogen-free, adult male Sprague-Dawley (SD) rats (180-230 g) were used in this study. Five rats were to be used for limb ischemia/reperfusion, then plasma was extracted as ischemia/reperfusion serum stimulation. Fifteen rats were to be used for extracting AM by bronchoalveolar lavage. The AM was isolated and cultured, then the cell count was adjusted to 1×10 6/mL, and randomly divided into the following 4 groups (n=6): control group, I/R group, SO2 group, and I/R+SO2 group. The I/R group was given ischemia/reperfusion serum (500 μg/L) to stimulate 6 h; the SO2 group was given an SO2 donor, Na2SO3/NaHSO3 [(0.54 mmol/kg) / (0.18 mmol/kg)]; and the I/R+SO2 group was given the same ischemia/reperfusion serum and Na2SO3/NaHSO3 at the same time. The level of mitochondrial membrane potential, the state of mitochondrial permeability transition pore (mPTP), the rate of AM apoptosis, the expression of Bcl-2 and Caspase-3 proteins were detected by flow cytometry, microplate reader and Western blotting.Results: Compared with the control group, in the I/R group, the ratio of red to green fluorescence and the absorbance decreased significantly, the percentage of apoptotic cells increased obviously, the apoptotic rate was 43.81%±2.40%, Caspase-3 protein expression increased, Bcl-2 protein expression decreased. While compared with the I/R group, in the I/R+SO2 group, the ratio of red to green fluorescence and the absorbance increased significantly; the apoptotic rate decreased to 37.01%±1.93%, Caspase-3 protein expression decreased, Bcl-2 protein expression increased.Conclusion: Exo-genous SO2 has the effect of accelerating AM apoptosis by stimulating mPTP to open and mitochondrial membrane potential to decrease; besides, exogenous SO2 could stimulate AM to secrete more anti-inflammatory cytokines and less inflammatory cytokines. In conclusion, exogenous SO2 can reduce macrophage apoptosis by inhibiting mitochondrial pathways.

Key words: Sulfur dioxide, Reperfusion injury, Acute lung injury, Macrophages, alveolar

中图分类号: 

  • R364.12

表1

SO2对AM线粒体膜电位及mPTP开放情况的影响(n=8)"

Group Ratio (Red/Green) Absorbance
Control 4.94±0.26 0.45±0.03
SO2 5.00±0.29 0.44±0.02
I/R 1.75±0.24* 0.24±0.02*
I/R+SO2 2.12±0.15# 0.27±0.03#

图1

Annexin V-FITC和PI双标流式细胞仪法检测AM凋亡情况"

图2

AM中Bcl-2、Caspase-3蛋白表达情况"

[1] Steiger AK, Yang Y, Royzen M , et al. Bio-orthogonal “click-and-release” donation of caged carbonyl sulfide (COS) and hydrogen sulfide (H2S)[J]. Chem Commun (Camb), 2017,53(8):1378-1380.
doi: 10.1039/C6CC09547J
[2] Steiger AK, Pardue S, Kevil CG , et al. Self-immolative thiocarbamates provide access to triggered H2S donors and analyte replacement fluorescent probes[J]. J Am Chem Soc, 2016,138(23):7256-7259.
doi: 10.1021/jacs.6b03780
[3] Aggarwal NR, D’Alessio FR, Tsushima K , et al. Moderate oxygen augments lipopolysaccharide-induced lung injury in mice[J]. Am J Physiol Lung Cell Mol Physiol, 2010,298(3):L371-L381.
doi: 10.1152/ajplung.00308.2009
[4] Howard KM . Differential expression of platelet-activating factor acetylhydrolase in lung macrophages[J]. Am J Physiol Lung Cell Mol Physiol, 2009,297(6):L1141-L1150.
doi: 10.1152/ajplung.00022.2009
[5] Z’Graggen BR, Tornic J, Muller-Edenborn B , et al. Acute lung injury: apoptosis in effector and target cells of the upper and lower airway compartment[J]. Clin Exp Immunol, 2010,161(2):324-331.
[6] Meng Z, Liu Y . Cell morphological ultrastructural changes in va-rious organs from mice exposed by inhalation to sulfur dioxide[J]. Inhal Toxicol, 2007,19(6-7):543-551.
doi: 10.1080/08958370701271373
[7] Ubuka T, Yuasa S, Ohta J , et al. Formation of sulfate from L-cysteine in rat liver mitochondria[J]. Acta Med Okayama, 1990,44(2):55-64.
[8] 赵彦瑞, 刘洋, 王东 , 等. PI3KAkt和JAK2STAT3信号转导通路在SO2抗大鼠肢体缺血再灌注致急性肺损伤中的作用[J]. 中华病理生理杂志, 2015,31(11):2076-2082.
[9] Huang XL, Liu Y, Zhou JL , et a1. Role of sulfur dioxide in acute lung injury following limb ischemia/reperfusion in rats[J]. J Biochem Mol Toxicol, 2013,27(8):389-397.
doi: 10.1002/jbt.2013.27.issue-8
[10] Zhao YR, Wang D, Liu Y , et al. The PI3K/Akt, p38MAPK, and JAK2/STAT3 signaling pathways mediate the protection of SO2 against acute lung injury induced by limb ischemia/reperfusion in rats[J]. J Physiol Sci, 2016,66(3):229-239.
doi: 10.1007/s12576-015-0418-z
[11] Cohen SM, Siddiqi FA, Darakchiev B , et al. Attenuation of acute lung injury caused by hind-limb ischemia-reperfusion injury by butyrolactone anti-inflammatory agent FLl003[J]. J Trauma, 1997,43(2):247-252.
doi: 10.1097/00005373-199708000-00007
[12] Javadov S, Choi A, Rajapurohitam V , et al. NHE-1 inhibition-induced cardioprotection against ischaemia/reperfusion is associa-ted with attenuation of the mitochondrial permeability transition[J]. Cardiovasc Res, 2008,77(2):416-424.
[13] Kaltenbach JP, Jennings RB . Metabolism of ischemic cardiac muscle[J]. Circ Res, 1960(8):207-213.
[14] Ward PA . Editorial commentary: New strategies for treatment of humans with acute lung injury/acute respiratory distress syndrome[J]. Clin Infect Dis, 2015,60(4):596-597.
doi: 10.1093/cid/ciu892
[15] Tauber AI . Metchnikoff and the phagocytosis theory[J]. Nat Rev Mol Cell Biol, 2003,4(11):897-901.
doi: 10.1038/nrm1244
[16] Ovchinnikov, Dmitry A . Macrophages in the embryo and beyond: Much more than just giant phagocytes[J]. Genesis, 2008,46(9):447-462.
doi: 10.1002/dvg.v46:9
[17] Yong Y, Matthew S, Wittwer J , et al. Dichamanetin inhibits cancer cell growth by affecting ROS-related signaling components through mitochondrial-mediated apoptosis[J]. Anticancer Res, 2013,33(12):5349-5355.
[1] 贾园,栗占国. 成人巨噬细胞活化综合征诊断困境和个体化治疗[J]. 北京大学学报（医学版）, 2020, 52(6): 991-994.
[2] 陈颖怡,胡紫琪,惠甜倩,刘鹤. Zeste同源蛋白2增强子通过调节巨噬细胞趋化影响牙髓炎症反应[J]. 北京大学学报（医学版）, 2020, 52(1): 18-23.
[3] 姚海红,王旖旎,张霞,赵金霞,贾园,王昭,栗占国. 67例成人巨噬细胞活化综合征的临床特征及治疗转归[J]. 北京大学学报（医学版）, 2019, 51(6): 996-1002.
[4] 王莹,李明慧,张岩,胡晓燕,马瑞霞. 狼疮性肾炎患者足细胞损伤与肾组织巨噬细胞浸润的关系[J]. 北京大学学报（医学版）, 2019, 51(4): 723-727.
[5] 吴小静，李敏，詹庆元. 以高热为表现的外源性脂质性肺炎1例[J]. 北京大学学报(医学版), 2018, 50(5): 921-923.
[6] 余建峰, 金月波, 何菁, 安媛, 栗占国. 皮肌炎继发干燥综合征伴肺间质病变的血清人Ⅱ型肺泡细胞表面抗原变化1例[J]. 北京大学学报(医学版), 2017, 49(5): 910-914.
[7] 高翔，陈香梅，张婷，张静，陈茉，郭正阳，石岩岩，鲁凤民，丁士刚. 巨噬细胞加帽蛋白与胃癌细胞增殖及迁移能力的关系[J]. 北京大学学报(医学版), 2017, 49(3): 489-494.
[8] 涂静宜, 朱莹, 尚淑玲, 张茜, 唐慧, 王瑞敏. Keap1-tat小肽降低缺血后大鼠海马CA1区神经元氧化应激损伤和空间学习记忆缺陷[J]. 北京大学学报(医学版), 2016, 48(1): 154-159.
[9] 李刚，张洪宪，王云鹏，张径，洪锴，田晓军，马潞林. 间苯三酚对大鼠肾缺血再灌注损伤的保护作用[J]. 北京大学学报(医学版), 2015, 47(5): 743-748.
[10] 王艳飞, 贾新未, 赵文萍, 王凤娟, 刘亚宁, 张芳, 张丽敏, 王鸿超. 辛伐他汀后适应对缺血再灌注损伤大鼠TNF-α及NF-κB的影响[J]. 北京大学学报（医学版）, 2014, 46(6): 990-992.
[11] 王宁, 王贵松, 于海奕, 米琳, 郭丽君, 高炜. 远隔缺血后适应在急性ST段抬高型心肌梗死直接经皮冠状动脉介入治疗术中的心肌保护作用[J]. 北京大学学报（医学版）, 2014, 46(6): 838-843.
[12] 赵曼曼, 张巧丽, 闫辉, 杜军保, 耿彬, 唐朝枢, 金红芳. H2S对氧化型低密度脂蛋白诱导人单核巨噬细胞核转录因子-κB的影响及机制[J]. 北京大学学报（医学版）, 2013, 45(2): 192-.
[13] 金红芳, 李真真. 内源性含硫氨基酸代谢终产物的心血管调节作用[J]. 北京大学学报（医学版）, 2013, 45(2): 177-.
[14] 濮吉, 陈田, 陈章健, 王海芳, 聂海瑜, 贾光. 比较不同长度及表面修饰的多壁碳纳米管的细胞毒性和遗传毒性[J]. 北京大学学报（医学版）, 2013, 45(03): 405-411.
[15] 纪翔, 马潞林, 卢剑, 张树栋, 黄毅, 侯小飞, 马鑫. 肾缺血后处理对大鼠Kim-1表达的影响及对缺血再灌注损伤的保护作用[J]. 北京大学学报（医学版）, 2012, 44(4): 511-517.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
[1] 田增民, 陈涛, Nanbert ZHONG, 李志超, 尹丰, 刘爽. 神经干细胞移植治疗遗传性小脑萎缩的临床研究（英文稿）[J]. 北京大学学报（医学版）, 2009, 41(4): 456 -458 .
[2] 郭岩, 谢铮. 用一代人时间弥合差距——健康社会决定因素理论及其国际经验[J]. 北京大学学报（医学版）, 2009, 41(2): 125 -128 .
[3] 成刚, 钱振华, 胡军. 艾滋病项目自愿咨询检测的技术效率分析[J]. 北京大学学报（医学版）, 2009, 41(2): 135 -140 .
[4] 卢恬, 朱晓辉, 柳世庆, 郑杰, 邱晓彦. 白细胞介素2促进宫颈癌细胞系HeLaS3免疫球蛋白G的表达[J]. 北京大学学报（医学版）, 2009, 41(2): 158 -161 .
[5] 袁惠燕, 张苑, 范田园. 离子交换型栓塞微球及其载平阳霉素的制备与性质研究[J]. 北京大学学报（医学版）, 2009, 41(2): 217 -220 .
[6] 徐莉, 孟焕新, 张立, 陈智滨, 冯向辉, 释栋. 侵袭性牙周炎患者血清中抗牙龈卟啉单胞菌的IgG抗体水平的研究[J]. 北京大学学报（医学版）, 2009, 41(1): 52 -55 .
[7] 董稳, 刘瑞昌, 刘克英, 关明, 杨旭东. 氯诺昔康和舒芬太尼用于颌面外科术后自控静脉镇痛的比较[J]. 北京大学学报（医学版）, 2009, 41(1): 109 -111 .
[8] 祁琨, 邓芙蓉, 郭新彪. 纳米二氧化钛颗粒对人肺成纤维细胞缝隙连接通讯的影响[J]. 北京大学学报（医学版）, 2009, 41(3): 297 -301 .
[9] Jian-wei GU, Emily YOUNG, Zhi-jun PAN, Kevan B. TUCKER, Megan SHPARAGO, Min HUANG, Amelia Purser BAILEY. SD大鼠长期高盐饮食可导致其高血压并改变肾细胞因子基因表达谱[J]. 北京大学学报（医学版）, 2009, 41(5): 505 -515 .
[10] 李宏亮*, 安卫红*, 赵扬玉, 朱曦. 妊娠合并高脂血症性胰腺炎行血液净化治疗1例[J]. 北京大学学报（医学版）, 2009, 41(5): 599 -601 .
ISSN 1671-167X
CN 11-4691/R
主管单位：中华人民共和国教育部
主办单位：北京大学
地址： 北京市海淀区学院路38号 北京大学医学部 《北京大学学报（医学版）》
邮编：100191
电话：010-82801551
Email： xbbjb2@bjmu.edu.cn

《北京大学学报（医学版）》
微信公众号
版权所有 © 2018 《北京大学学报（医学版）》编辑部