[1]杨诚忠,李满满,罗羽莎,等.LIGHT在低氧性肺动脉高压形成中的作用及机制[J].第三军医大学学报,2018,40(08):643-651.
 YANG Chengzhong,LI Manman,LUO Yusha,et al.Role of LIGHT in development of hypoxic pulmonary hypertension in mice[J].J Third Mil Med Univ,2018,40(08):643-651.
点击复制

LIGHT在低氧性肺动脉高压形成中的作用及机制(/HTML )
分享到:

《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
40卷
期数:
2018年第08期
页码:
643-651
栏目:
基础医学
出版日期:
2018-04-30

文章信息/Info

Title:
Role of LIGHT in  development of hypoxic pulmonary hypertension in mice
作者:
杨诚忠李满满罗羽莎徐刚李桂清许桂莲谭小玲高钰琪
陆军军医大学(第三军医大学):高原军事医学系,高原生理学与病理学教研室,高原军事医学系,高原特殊药品与装备研究室,全军高原医学重点实验室,基础医学院免疫学教研室
 
Author(s):
YANG Chengzhong LI Manman LUO Yusha1 XU Gang LI Guiqing XU Guilian TAN Xiaoling GAO Yuqi

Department of High Altitude Physiology and Pathology, 2Department of High Altitude Special Drug and Equipment Research, Key Laboratory of High Altitude Medicine of PLA, College of High Altitude Military Medicine, 3Department of Immunology, College of Basic Medical Sciences, Army Medical University (Third Military Medical University) , Chongqing, 400038, China

关键词:
LIGHTIL-6单核巨噬细胞肺动脉高压低氧
Keywords:
tumor necrosis factor superfamily member 14 interleukin-6 monocyte-macrophages pulmonary hypertension hypoxia
分类号:
R392.11;R363.21;R544.02
文献标志码:
A
摘要:

目的     初步探讨LIGHT在低氧性肺动脉高压(hypoxic pulmonary hypertension, HPH)形成中的作用及其机制。方法     将20只8周龄雌性C57BL/6J小鼠[体质量(17.90±0.91)g]和20只8周龄雌性LIGHT-/- C57BL/6J小鼠[体质量(17.55±0.93)g]分为4组(n=10):①野生小鼠常氧组(WT-C 组)、②野生小鼠低氧组(WT-H组)、③LIGHT KO小鼠常氧组(LIGHT KO-C组)、④LIGHT KO小鼠低氧组(LIGHT KO-H组)。WT-H组和LIGHT KO-H组小鼠置于模拟6 000 m低压舱内连续低氧饲养30 d,WT-C组和LIGHT KO-C组小鼠舱外(海拔308 m)常规饲养。检测右心室收缩压(right ventricular systolic pressure,RVSP)和右心肥厚指数(right ventricular hypertrophy index,RVHI);HE染色观察肺小动脉结构;免疫组化检测LIGHT及其受体HVEM、LTβR表达;荧光定量PCR和Western blot检测肺组织LIGHT、HVEM和LTβR、IL-6的mRNA和蛋白水平。流式细胞术检测肺组织中各类炎症细胞的比例。结果     与WT-C组相比,WT-H组肺组织中LIGHT的mRNA和蛋白水平均显著增高(P<0.05),WT-H组RVSP和RVHI明显升高(P<0.05),肺小动脉明显增厚;与LIGHT KO-C组相比,LIGHT KOH组小鼠的RVSP、RVHI和肺小动脉厚度显著增加(P<0.05),但与WT-H组相比,LIGHT KO-H组的RVSP、RVHI和肺小动脉增厚程度明显降低(P<0.05)。与WTC组相比,WT-H组LIGHT受体HVEM表达增加,LTβR表达降低,差异具有统计学意义(P<0.05)。LIGHT KOH组与WT-H组相比,肺组织IL-6 mRNA和蛋白水平显著降低(P<0.05)。流式细胞检测发现,与WTC组相比,WTH组小鼠肺组织中单核细胞比例降低[(3.88±0.87)% vs (11.03±1.71)%,P<0.05 ],间质巨噬细胞比例升高[(15.56±2.69)% vs (8.57±2.17)%,P<0.05 ];与WTH组相比,LIGHT KOH组的肺组织单核细胞增加[(6.55±1.01)% vs (3.88±0.87)%,P<0.05 ],而间质巨噬细胞的比例降低[(10.87±1.68)% vs.(15.56±2.69)%,P<0.05 ]。结论    慢性低氧诱导肺组织中LIGHT表达增加与HPH发病机制密切相关。LIGHT可能通过HVEM信号途径促进细胞增殖、上调肺组织IL-6表达、促进肺间质巨噬细胞产生,参与HPH的形成。
 

Abstract:

Objective     To investigate the role of tumor necrosis factor superfamily member 14 (LIGHT/TNFSF14) in the development of hypoxic pulmonary hypertension in mice and explore the mechanism. Methods     Twenty 8-week-old wild-type (WT) female C57BL/6J mice and 20 age-matched LIGHT-deficient (LIGHT-/-) transgenic female C57BL/6J mice were both randomized equally into normoxia groups and hypoxia groups. In the 2 hypoxia groups, the mice were housed in a hypobaric chamber simulating the condition of 6 000 m altitude for 30 d, and mice in normoxic groups were housed in routine conditions. The changes in right ventricular systolic pressure (RVSP) were recorded and right ventricular hypertrophy index (RVHI) was evaluated. HE staining was used to observe the changes in pulmonary vascular structures. Immunohistochemical staining was performed to observe the distribution of LIGHT, HVEM and LTβR in the lung of the mice. The expressions of LIGHT, HVEM, LTβR and IL-6 at both mRNA and protein levels in the lungs were tested using RT-PCR and Western blotting. The proportions of inflammatory cells in the lungs were determined using flow cytometry. Results     Compared with the WT normoxic mice, the WT mice with chronic hypoxia showed significantly increased mRNA and protein levels of LIGHT in the lungs (P<0.05) with also significantly increased RVSP and RVHI and obviously thickened pulmonary arterioles (P<0.05). These hypoxiainduced changes were markedly in attenuated LIGHT-/-  mice. In WT mice, hypoxia induced significantly increased protein expression of LIGHT receptor HVEM (P<0.05) and decreased expression of LTβR (P<0.05). LIGHT-/- mice with hypoxia showed significantly lower mRNA and protein levels of IL-6 in the lungs than WT hypoxic mice (P<0.05). Compared with WT normoxic mice, WT hypoxic mice showed significantly decreased proportions of monocytes [(3.88±0.87)% vs (11.03±1.71)%, P<0.05 ] and increased interstitial macrophages [(15.56±2.69)% vs (8.57±2.17)%, P<0.05 ] in the lungs. In LIGHT-/- mice with hypoxia, the proportion of monocytes was significantly increased to (6.55±1.01)% (P<0.05) and that of interstitial macrophage was significantly decreased to (10.87±1.68)% (P<0.05) compared with those in WT hypoxic mice. Conclusion    Chronic hypoxia-induced development of HPH is closely associated with increased pulmonary LIGHT expression, which promotes cell growth, upregulates IL-6, and increases the production of pulmonary interstitial macrophages possibly by activating HVEM under hypoxic condition.

参考文献/References:

[1]MCLAUGHLIN V V, ARCHER SL, BADESCH D B, et al. ACCF/AHA 2009 expert consensus document on pulmonary hypertension a report of the American College of Cardiology Foundation Task Force on Expert Consensus Documents and the American Heart Association developed in collaboration with the American College of Chest Physicians;American Thoracic Society, Inc.; and the Pulmonary Hypertension Association[J]. J Am Coll Cardiol, 2009. 53(17): 1573-619. DOI: 10.1016/j.jacc.2009.01.004
[2]PUGLIESE SC, POTH JM,  FINI MA,  et al. The role of inflammation in hypoxic pulmonary hypertension: from cellular mechanisms to clinical phenotypes[J]. Am J Physiol Lung Cell Mol Physiol, 2015. 308(3): L229-52. DOI: 10.1152/ajplung.00238.2014
[3]HALVORSEN B, SANTILLI F, SCHOLZ H,  et al. LIGHT/TNFSF14 is increased in patients with type 2 diabetes mellitus and promotes islet cell dysfunction and endothelial cell inflammation in vitro[J]. Diabetologia, 2016. 59(10): 2134-2144. DOI: 10.1007/s001250164036y
[4]KRAUSE P, ZAHNER S P,  KIM G, et al. The tumor necrosis factor family member TNFSF14 (LIGHT) is required for resolution of intestinal inflammation in mice[J]. Gastroenterology,  2014. 146(7): 1752-1762 e4. DOI: 10.1053/j.gastro.2014.02.010
[5]DOHERTY T A, SOROOSH P, KHORRAM N, et al. The tumor necrosis factor family member LIGHT is a target for asthmatic airway remodeling[J]. Nat Med, 2011. 17(5): 596-603. DOI:  10.1038/nm.2356
[6]OTTERDAL K, ANDREASSEN AK, YNDESTAD A, et al. Raised LIGHT levels in pulmonary arterial hypertension: potential role in thrombus formation[J]. Am J Respir Crit Care Med,  2008. 177(2): 202-207. DOI:  10.1164/rccm.200703506OC
[7]BECHILL J, MULLER W J. Herpesvirus entry mediator (HVEM) attenuates signals mediated by the lymphotoxin beta receptor (LTbetaR) in human cells stimulated by the shared ligand LIGHT[J]. Mol Immunol, 2014. 62(1): 96-103. DOI:  10.1016/j.molimm.2014.06.013
[8]FLORENTIN J, DUTTA P. Origin and production of inflammatory perivascular macrophages in pulmonary hypertension[J]. Cytokine, 2017. 100: 11-15. DOI: 10.1016/j.cyto.2017.08.015
[9]WEI C Y, CHOU Y H, HO F M, et al. Signaling pathways of LIGHT induced macrophage migration and vascular smooth muscle cell proliferation[J]. J Cell Physiol, 2006. 209(3): 735-743. DOI: 10.1002/jcp.20742
[10]FRID M G, BRUNETTI J A, BURKE D L, et al. Hypoxiainduced pulmonary vascular remodeling requires recruitment of circulating mesenchymal precursors of a monocyte/macrophage lineage[J]. Am J Pathol, 2006. 168(2): 659-669. DOI:  10.2353/ajpath.2006.050599
[11]CHEN T, YANG C, LI M, et al. Alveolar HypoxiaInduced Pulmonary Inflammation: From Local Initiation to Secondary Promotion by Activated Systemic Inflammation[J]. J Vasc Res, 2016. 53(5-6): 317-329. DOI:  10.1159/000452800
[12]BURKE D L, FRID M G, KUNRATH C L, et al. Sustained hypoxia promotes the development of a pulmonary arteryspecific chronic inflammatory microenvironment[J]. Am J Physiol Lung Cell Mol Physiol, 2009. 297(2): L238-250. DOI:  10.1152/ajplung.90591.2008
[13]BEDORET D, WALLEMACQ H, MARICHAL T, et al. Lung interstitial macrophages alter dendritic cell functions to prevent airway allergy in mice[J]. J Clin Invest, 2009. 119(12): 3723-3738. DOI:  10.1172/JCI39717
[14]GIBBINGS S L, THOMAS S M, ATIF S M, et al. Three Unique Interstitial Macrophages in the Murine Lung at Steady State[J]. Am J Respir Cell Mol Biol, 2017. 57(1): 66-76. DOI:  10.1165/rcmb.20160361OC
[15]GUILLIAMS M, DE KLEER I, HENRI S, et al. Alveolar macrophages develop from fetal monocytes that differentiate into longlived cells in the first week of life via GMCSF[J]. J Exp Med, 2013. 210(10): 1977-1992. DOI:  10.1084/jem.20131199
[16]PUGLIESE S C, KUMAR S, JANSSEN W J, et al. A Time and CompartmentSpecific Activation of Lung Macrophages in Hypoxic Pulmonary Hypertension[J]. J Immunol, 2017. 198(12): 4802-4812. DOI:  10.4049/jimmunol.1601692
[17]HASHIMOTOKATAOKA T, HOSEN N, SONOBE T, et al. Interleukin6/interleukin21 signaling axis is critical in the pathogenesis of pulmonary arterial hypertension[J]. Proc Natl Acad Sci U S A, 2015. 112(20): E2677-2686. DOI: 10.1073/pnas.1424774112
[18]MASTON L D, JONES D T, GIERMAKOWSKA W, et al. Central role of T helper 17 cells in chronic hypoxia-induced pulmonary hypertension[J]. Am J Physiol Lung Cell Mol Physiol, 2017. 312(5): L609-L624. DOI:  10.1152/ajplung.00531.2016
[19]SAVALE L, TU L, RIDEAU D, et al. Impact of interleukin-6 on hypoxia-induced pulmonary hypertension and lung inflammation in mice[J]. Respir Res, 2009. 10: 6. DOI:  10.1186/1465-9921-106
[20]STEINER  M K, SYRKINA O L, KOLLIPUTI N, et al. Interleukin6 overexpression induces pulmonary hypertension[J]. Circ Res, 2009. 104(2): 236-244, 28p following 244. DOI:  10.1161/CIRCRESAHA.108.182014
[21]KAWANO H, KAYAMA H, NAKAMA T, et al. IL-10-producing lung interstitial macrophages prevent neutrophilic asthma[J]. Int Immunol, 2016. 28(10): 489-501. DOI:  10.1093/intimm/dxw012

相似文献/References:

[1]臧春宝,曹姝,李芳,等.IL-6对食管癌细胞侵袭迁移及上皮间质转化的影响[J].第三军医大学学报,2013,35(04):302.
 Zang Chunbao,Cao Shu,Li Fang,et al.Effects of interleukin-6 on invasion, migration and epithelial-mesenchymal transition in human esophageal carcinoma cells[J].J Third Mil Med Univ,2013,35(08):302.
[2]刘占国,谭晓莹,蔡靓,等.脓毒症患者危重程度与血清IL-6的相关性分析[J].第三军医大学学报,2012,34(21):2233.
[3]任泂,粟永萍,李洪涛,等.JAB1表达下调对LPS诱导炎性因子TNF-α和IL-6的影响[J].第三军医大学学报,2007,29(16):1552.
 REN Jiong,SU Yong-ping,LI Hong-tao,et al.Effect of JAB1 down-regulation on LPS-induced TNF-α and IL-6 in mouse macrophage cells[J].J Third Mil Med Univ,2007,29(08):1552.
[4]韩京,冉新泽,粟永萍,等.辐射损伤对巨核细胞造血调控因子IL-6和TNF-α的影响[J].第三军医大学学报,2005,27(18):1817.
[5]张晓东,杨天德,李洪.星状神经节阻滞对体外循环术中皮质醇、TNF-α、IL-1β、IL-6的影响[J].第三军医大学学报,2010,32(10):1028.
 Zhang Xiaodong,Yang Tiande,Li Hong.Effect of stellate ganglion blocking on serum levels of Col, TNF-α, IL-1β and IL-6 in patients during cardiopulmonary bypass[J].J Third Mil Med Univ,2010,32(08):1028.
[6]董晓灵,王曙光,张玉君,等.VEGF对大鼠50%肝脏移植术后血清HGF、IL-6浓度的影响[J].第三军医大学学报,2008,30(11):1037.
 DONG Xiao-ling,WANG Shu-guang,ZHANG Yu-jun,et al.VEGF enhances serum levels of HGF and IL-6 in rats following 50 percent partial liver transplantation[J].J Third Mil Med Univ,2008,30(08):1037.
[7]赵永亮,余佩武,刘伟,等.IL-6通过SOCS3诱导小鼠髓源性DCs分化成熟障碍的分子机制研究[J].第三军医大学学报,2011,33(15):1568.
 Zhao Yongliang,Yu Peiwu,Liu Wei,et al.IL-6 induces disturbance on differentiation and maturation of mouse bone marrow derived dendritic cells through SOCS3[J].J Third Mil Med Univ,2011,33(08):1568.
[8]黄艳玲,秦新月,王恬竹,等.马来酸桂哌齐特对大鼠局灶性脑缺血再灌注后白介素-1β、白介素-6的表达及神经功能的影响[J].第三军医大学学报,2011,33(06):596.
 Huang Yanling,Qin Xinyue,Wang Tianzhu,et al.Effects of cinepazide maleate on IL-1β and IL-6 expression and neural function in rats after focal cerebral ischemia/reperfusion[J].J Third Mil Med Univ,2011,33(08):596.
[9]倪志宇,丛斌,董玫,等.八肽胆囊收缩素对脂多糖诱导RAW 264.7细胞IL-1β、IL-6、IL-10表达的影响[J].第三军医大学学报,2011,33(22):2366.
 Ni Zhiyu,Cong Bin,Dong Mei,et al.Effect of cholecystokinin octapeptide on IL-1β, IL-6 and IL-10 expression in LPS-induced RAW 264.7 cells[J].J Third Mil Med Univ,2011,33(08):2366.
[10]吴晓云,匡凤梧,吴春,等.小儿体外循环围术期炎症因子释放规律及机制探讨[J].第三军医大学学报,2005,27(08):776.

更新日期/Last Update: 2018-04-25