超细颗粒物对大鼠离体灌注心脏功能的影响

doi:10.19723/j.issn.1671-167X.2021.02.002

摘要/Abstract

摘要：

目的: 研究超细颗粒物(ultrafine particulates,UFPs)对大鼠离体心脏功能的影响及其机制。方法: 以含或不含UFPs的台式液经Langendorff系统持续灌流大鼠离体心脏模型40 min,观察灌流前后两组大鼠心脏血流动力学指标[左心室舒张压(left ventricular developed pressure,LVDP)、左心室内压最大上升和最大下降速率(±dp/dt_max)及冠脉流量(coronary flow,CF)]的变化。收集肺动脉流出液,采用硫代巴比妥酸法测定丙二醛(malondialdehyde,MDA),水溶性四唑盐法测定超氧化物歧化酶(superoxide dismutase,SOD), 比色法测定总抗氧化能力(total antioxidant capacity,TAOC)。免疫组织化学法和Western blots法测定两组心脏标本p-p38 MAPK、p-JNKs、p-ERKs的表达。结果: 相对于对照组,UFPs灌流组大鼠离体心脏功能指标LVDP、+dp/dt_max、-dp/dt_max、CF分别从(82.6 ± 2.1) mmHg、(1 624 ± 113) mmHg/s、(1 565 ± 116) mmHg/s、(12.0 ± 0.2) mL/min降至灌注结束时的(56.8 ± 4.4) mmHg、(1 066 ± 177) mmHg/s、(1 082 ± 134) mmHg/s、(8.7 ± 0.3) mL/min,各指标灌流结束时相比灌流初始值差异均有统计学意义(P<0.05)。UFPs灌流组肺动脉流出液MDA含量明显高于对照组[(1.95±0.18) nmol/L vs. (0.98±0.14) nmol/L,P<0.05],而SOD、TAOC明显低于对照组[(6.50±1.04) U/mL vs. (12.50±1.87) U/mL,(3.67±0.82) U/mL vs. (6.83±1.16) U/mL, P<0.05 ]。UFPs灌流组p-p38 MAPK、p-JNKs、p-ERKs 较对照组表达明显增加(P<0.05)。结论: UFPs短期暴露对大鼠离体心脏有直接的急性毒性作用,其作用机制可能与氧化应激及MAPK信号通路激活有关。

关键词: 超细颗粒物, 大鼠离体心脏, 氧化应激, MAPK

Abstract:

Objective: To evaluate whether ultrafine particulates (UFPs) have direct deleterious effects on cardiac function through activating MAPK signaling. Methods: Langendorff-perfused Sprague-Dawley rat hearts were randomly divided into 2 groups (n=10/each group). In control group, the rat hearts were perfused with Tyrode’s buffer for 40 min; in UFPs-treated group, the hearts were perfused with UFPs at a concentration of 12.5 mg/L. Cardiac function was determined by measuring left ventricular developed pressure (LVDP), left ventricular peak rate of contraction and relaxation (±dp/dt_max) and coronary flow (CF). The levels of malondialdehyde (MDA), superoxide dismutase (SOD), total anti-oxidant capacity (TAOC) were detected in order to evaluate cardiac oxidative stress via the thiobarbituric acid assay, water soluble tetrazolium salt assay and colorimetry, respectively. The expressions of p-p38 MAPK, p-ERKs and p-JNKs in the myocardium were observed using immunohistochemical staining and Western blots. Results: No significant changes in cardiac function were detected before and after the perfusion in control group while UFPs perfused hearts showed a decline in cardiac function in a time-dependent manner (all P<0.05). In UFPs-treated group, LVDP, +dp/dt_max, -dp/dt_max and CF were statistically reduced from (82.6±2.1) mmHg, (1 624±113) mmHg/s, (1 565±116) mmHg/s, (12.0±0.2) mL/min to (56.8±4.4) mmHg, (1 066±177) mmHg/s, (1 082±134) mmHg/s, (8.7±0.3) mL/min (all P<0.05), respectively. Furthermore, The comparison between the two groups observed that UFPs perfusion caused a significant decrease in cardiac function at 30 and 40 min compared with the control group (all P<0.05). At the end of the perfusion, the level of MDA was increased from (0.98±0.14) nmol/L to (1.95±0.18) nmol/L, while SOD and TAOC were reduced from (12.50±1.87) U/mL and (6.83±1.16) U/mL to (6.50 ±1.04) U/mL and (3.67±0.82) U/mL (all P<0.001) in UFPs group, respectively. In coincidence with these changes, immunohistochemistry and Western blots results showed that the levels of p-p38 MAPK, p-ERKs and p-JNKs in the myocardium significantly increased in UFPs group as compared with control group (all P<0.05). Conclusion: The results of this study demonstrated that the short-term exposure of UFPs to the isolated rat hearts has direct and acute toxic effects on cardiac function, probably related to attenuation of anti-oxidative capacity and activation of MAPK signaling pathways.

Key words: Ultrafine particulates(UFPs), Isolated rat heart, Oxidative stress, MAPK

中图分类号:

R122.7

白枫,何倚帆,牛亚楠,杨若娟,曹静. 超细颗粒物对大鼠离体灌注心脏功能的影响[J]. 北京大学学报（医学版）, 2021, 53(2): 240-245.

BAI Feng,HE Yi-fan,NIU Ya-nan,YANG Ruo-juan,CAO Jing. Effects of ultrafine particulates on cardiac function in rat isolated heart[J]. Journal of Peking University (Health Sciences), 2021, 53(2): 240-245.

图/表 5

图1

表1

图2

图3

图4

参考文献 19

[1]	Klompmaker JO, Montagne DR, Meliefste K, et al. Spatial variation of ultrafine particles and black carbon in two cities: results from a short-term measurement campaign[J]. Sci Total Environ, 2015,508:266-275.
[2]	Zareba W, Couderc JP, Oberdörster G, et al. ECG parameters and exposure to carbon ultrafine particles in young healthy subjects[J]. Inhal Toxicol, 2009,21(3):223-233. doi: 10.1080/08958370802492407 pmid: 18991063
[3]	Laumbach RJ, Kipen HM, Ko S, et al. A controlled trial of acute effects of human exposure to traffic particles on pulmonary oxidative stress and heart rate variability[J]. Part Fibre Toxicol, 2014,11:45. pmid: 25361615
[4]	Mills NL, Amin N, Robinson SD, et al. Do inhaled carbon nanoparticles translocate directly into the circulation in humans[J]. Am J Respir Crit Care Med, 2006,173(4):426-431. pmid: 16339922
[5]	Lundbäck M, Mills NL, Lucking A, et al. Experimental exposure to diesel exhaust increases arterial stiffness in man[J]. Part Fibre Toxicol, 2009,6:7. pmid: 19284640
[6]	Wang M, Beelen R, Stafoggia M, et al. Long-term exposure to elemental constituents of particulate matter and cardiovascular mortality in 19 European cohorts: results from the ESCAPE and TRANSPHORM projects[J]. Environ Int, 2014,66:97-106. doi: 10.1016/j.envint.2014.01.026 pmid: 24561271
[7]	Stewart JC, Chalupa DC, Devlin RB, et al. Vascular effects of ultrafine particles in persons with type 2 diabetes[J]. Environ Health Perspect, 2010,118(12):1692-1698. pmid: 20822968
[8]	Nemmar A, Subramaniyan D, Yasin J, et al. Impact of experimental type 1 diabetes mellitus on systemic and coagulation vulnerability in mice acutely exposed to diesel exhaust particles[J]. Part Fibre Toxicol, 2013,10:14. pmid: 23587270
[9]	Rückerl R, Phipps RP, Schneider A, et al. Ultrafine particles and platelet activation in patients with coronary heart disease: results from a prospective panel study[J]. Part Fibre Toxicol, 2007,4:1. doi: 10.1186/1743-8977-4-1 pmid: 17241467
[10]	Sun Q, Yue P, Ying Z, et al. Air pollution exposure potentiates hypertension through reactive oxygen species-mediated activation of Rho/ROCK[J]. Arterioscler Thromb Vasc Biol, 2008,28(10):1760-1766. pmid: 18599801
[11]	Simkhovich BZ, Marjoram P, Kleinman MT, et al. Direct and acute cardiotoxicity of ultrafine particles in young adult and old rat hearts[J]. Basic Res Cardiol, 2007,102(6):467-475.
[12]	Shaw CA, Robertson S, Miller MR, et al. Diesel exhaust particulate: exposed macrophages cause marked endothelial cell activation[J]. Am J Respir Cell Mol Biol, 2011,44(6):840-851.
[13]	Kim JB, Kim C, Choi E, et al. Particulate air pollution induces arrhythmia via oxidative stress and calcium calmodulin kinase Ⅱ activation[J]. Toxicol Appl Pharmacol, 2012,259(1):66-73.
[14]	Cozzi E, Hazarika S, Stallings HW, et al. Ultrafine particulate matter exposure augments ischemia-reperfusion injury in mice[J]. Am J Physiol Heart Circ Physiol, 2006,291(2):H894-903. pmid: 16582015
[15]	Cao J, Qin G, Shi RZ, et al. Overproduction of reactive oxygen species and activation of MAPKs are involved in apoptosis induced by PM2.5 in rat cardiac H9C2 cells[J]. J Apple Tocicol, 2016,36(4):609-617.
[16]	Baines CP, Molkentin JD. STRESS signaling pathways that modulate cardiac myocyte apoptosis[J]. J Mol Cell Cardiol, 2005,38(1):47-62. doi: 10.1016/j.yjmcc.2004.11.004 pmid: 15623421
[17]	Zhu W, Zou Y, Aikawa R, et al. MAPK superfamily plays an important role in daunomycin-induced apoptosis of cardiac myocytes[J]. Circulation, 1999,100(20):2100-2107. pmid: 10562267
[18]	Jarvis IW, Bergvall C, Morales DA, et al. Nanomolar levels of PAHs in extracts from urban air induce MAPK signaling in HepG2 cells[J]. Toxicol Lett, 2014,229(1):25-32. pmid: 24910982
[19]	Rui W, Guan L, Zhang F, et al. PM2.5-induced oxidative stress increases adhesion molecules expression in human endothelial cells through the ERK/AKT/NF-κB-dependent pathway[J]. J Appl Toxicol, 2016,36(1):48-59. pmid: 25876056

Metrics

Viewed

Full text

Abstract

Cited

Shared

Discussed