Acute exercise in ozone-polluted air induces apoptosis in rat quadriceps femoris muscle cells via mitochondrial pathway

Abstract

Ozone (O₃) pollution can decrease sport performance and induce respiratory toxicity, but relatively few studies have investigated its effects on skeletal muscles. We randomly assigned rats to the following groups based on a 2 ​× ​4 two-factor factorial design: Air+0, Air+10, Air+15, and Air+20, O₃+0, O₃+10, O₃+15, and O₃+20. The rats in the +0 groups rested, whereas those in the +10, +15, and +20 groups ran on a treadmill (in clean air for Air groups and in air polluted with 0.14 ​parts per million [ppm] O₃ for O₃ groups) at speeds of 10, 15, and 20 ​m/min, respectively, for 1 ​h. Thereafter, key enzyme activities involving the tricarboxylic acid cycle, oxidative phosphorylation, adenosine triphosphate (ATP) content, histopathological changes, oxidative stress, inflammation factors, and apoptosis were assessed in the rat quadriceps femoris samples. Ozone reduced key enzyme activities and ATP contents in the quadriceps femoris regardless of whether the rats exercised. Pathological changes, inflammatory factors, oxidative stress, and mitochondria-dependent apoptosis were only evident under conditions of exercise combined with ozone and increasingly worsened as exercise intensity increased. These findings suggested that acute exercise under ozone exposure could induce damage to the quadriceps femoris, which would negatively affect sport performance. Ozone-induced disrupted energy metabolism might be an early event that becomes more critical as exercise intensity increases. Therefore, care should be taken when exercising in polluted air, even when ozone pollution is mild.

Keywords

Ozone / Exercise / Quadriceps femoris / Apoptosis / Mitochondria

References

[[1]] RD Peng, E Samoli, L Pham, et al.. Acute effects of ambient ozone on mortality in Europe and North America: results from the APHENA study. Air Qual Atmos Health, 6 (2) ( 2013), pp. 445-453, DOI: 10.1007/s11869-012-0180-9

[[2]] R Chen, J Cai, X Meng, et al.. Ozone and daily mortality rate in 21 cities of East Asia: how does season modify the association?. Am J Epidemiol, 180 (7) ( 2014), pp. 729-736, DOI: 10.1093/aje/kwu183

[[3]] AJ Cohen, M Brauer, R Burnett, et al.. Estimates and 25-year trends of the global burden of disease attributable to ambient air pollution: an analysis of data from the Global Burden of Diseases Study 2015. Lancet, 389

( 10082 (2017), pp. 1907-1918, DOI: 10.1016/S0140-6736(17 ) 30505-6

[[4]] P Orellano, J Reynoso, N Quaranta, A Bardach, A Ciapponi.Short-term exposure to particulate matter (PM10 and PM2.5), nitrogen dioxide ( NO2), and ozone (O3) and all-cause and cause-specific mortality: systematic review and meta-analysis. Environ Int, 142 ( 2020), Article 105876, DOI: 10.1016/j.envint.2020.105876

[[5]] SK Sahu, S Liu, S Liu, D Ding, J Xing. Ozone pollution in China: background and transboundary contributions to ozone concentration & related health effects across the country. Sci Total Environ, 761 ( 2020), Article 144131, DOI: 10.1016/j.scitotenv.2020.144131

[[6]] Y Wang, K Wang, W Cheng, Y Zhang. Global burden of chronic obstructive pulmonary disease attributable to ambient ozone in 204 countries and territories during 1990-2019. Environ Sci Pollut Res Int, 29 (6) ( 2022), pp. 9293-9305, DOI: 10.1007/s11356-021-16233-y

[[7]] XY Zheng, P Orellano, HL Lin, M Jiang, WJ Guan. Short-term exposure to ozone, nitrogen dioxide, and sulphur dioxide and emergency department visits and hospital admissions due to asthma: a systematic review and meta-analysis. Environ Int, 150 (7) ( 2021), Article 106435, DOI: 10.1016/j.envint.2021.106435

[[8]] JJ Zhang, Y Wei, Z Fang.Ozone pollution: a major health hazard worldwide. Front Immunol, 10 ( 2019), p. 2518, DOI: 10.3389/fimmu.2019.02518

[[9]] WF Mcdonnell, PW Stewart, MV Smith, CS Kim, ES Schelegle. Prediction of lung function response for populations exposed to a wide range of ozone conditions. Inhal Toxicol, 24 (10) ( 2012), pp. 619-633, DOI: 10.3109/08958378.2012.705919

[[10]] EI Thaller, SA Petronella, D Hochman, et al.. Moderate increases in ambient PM2.5 and ozone are associated with lung function decreases in beach lifeguards. J Occup Environ Med, 50 (2) ( 2008), pp. 202-211, DOI: 10.1097/JOM.0b013e31816386b4

[[11]] GC Chuang, Z Yang, DG Westbrook, et al.. Pulmonary ozone exposure induces vascular dysfunction, mitochondrial damage, and atherogenesis. Am J Physiol Lung Cell Mol Physiol, 297 (2) ( 2009), pp. L209-L216, DOI: 10.1152/ajplung.00102.2009

[[12]] G Wang, R Jiang, Z Zhao, W Song. Effects of ozone and fine particulate matter (PM(2.5)) on rat system inflammation and cardiac function. Toxicol Lett, 217 (1) ( 2013), pp. 23-33, DOI: 10.1016/j.toxlet.2012.11.009

[[13]] L Tian, N Chu, H Yang, et al.. Acute ozone exposure can cause cardiotoxicity: mitochondria play an important role in mediating myocardial apoptosis. Chemosphere, 268 ( 2020), Article 128838, DOI: 10.1016/j.chemosphere.2020.128838

[[14]] RB Devlin, KE Duncan, M Jardim, et al.. Controlled exposure of healthy young volunteers to ozone causes cardiovascular effects. Circulation, 126 (1) ( 2012), pp. 104-111, DOI: 10.1161/CIRCULATIONAHA.112.094359

[[15]] X Jia, X Song, M Shima, et al.. Acute effect of ambient ozone on heart rate variability in healthy elderly subjects. J Expo Sci Environ Epidemiol, 21 (5) ( 2011), pp. 541-547, DOI: 10.1038/jes.2011.18

[[16]] WJ Gauderman, E Avol, F Gilliland, et al.. The effect of air pollution on lung development from 10 to 18 years of age. N Engl J Med, 351 (11) ( 2004), pp. 1057-1067, DOI: 10.1056/NEJMoa040610

[[17]] WJ Gauderman, et al..GF Gilliland, H Vora, Association between air pollution and lung function growth in southern California children: results from a second cohort. Am J Respir Crit Care Med, 166 (1) ( 2002), pp. 76-84, DOI: 10.1164/rccm.2111021

[[18]] N Künzli, M Jerrett, WJ Mack, et al.. Ambient air pollution and atherosclerosis in Los Angeles. Environ Health Perspect, 113 (2) ( 2005), pp. 201-206, DOI: 10.1289/ehp.7523

[[19]] IB Tager, J Balmes, F Lurmann, et al.. Chronic exposure to ambient ozone and lung function in young adults. Epidemiology, 16 (6) ( 2005), pp. 751-759, DOI: 10.1097/01.ede.0000183166.68809.b0

[[20]] YL Li, TW Chuang, PY Chang, et al.. Long-term exposure to ozone and sulfur dioxide increases the incidence of type 2 diabetes mellitus among aged 30 to 50 adult population. Environ Res, 194 (2) ( 2021), Article 110624, DOI: 10.1016/j.envres.2020.110624

[[21]] Email SA. Integrated Science Assessment (ISA) of Ozone and Related Photochemical Oxidants (Final Report, Feb 2013). United States Environmental Protection Agency; 2013.

[[22]] EJ Werder, LS Engel, KG Lawrence, DP Sandler.Ambient particulate matter, ozone, and neurologic symptoms in U.S. Gulf states adults. Environ Epidemiol, 5 (4) ( 2021), p. e160, DOI: 10.1097/EE9.0000000000000160

[[23]] GC Rowe, A Safdar, Z Arany. Running forward: new frontiers in endurance exercise biology. Circulation, 129 (7) ( 2014), pp. 798-810, DOI: 10.1161/CIRCULATIONAHA.113.001590

[[24]] JT Mullins. Ambient air pollution and human performance: contemporaneous and acclimatization effects of ozone exposure on athletic performance. Health Econ, 27 (8) ( 2018), pp. 1189-1200, DOI: 10.1002/hec.3667

[[25]] E Pierson. Impact of air pollutants on athletic performance. Allergy Proc, 10 (3) ( 1989), pp. 209-214, DOI: 10.2500/108854189778960144

[[26]] S Cakmak, R Dales, J Leech, L Liu. The influence of air pollution on cardiovascular and pulmonary function and exercise capacity: Canadian Health Measures Survey (CHMS). Environ Res, 111 (8) ( 2011), pp. 1309-1312, DOI: 10.1016/j.envres.2011.09.016

[[27]] EC Gomes, JE Allgrove, G Florida-James, V Stone. Effect of vitamin supplementation on lung injury and running performance in a hot, humid, and ozone-polluted environment. Scand J Med Sci Sports, 21 (6) ( 2011), pp. e452-e460, DOI: 10.1111/j.1600-0838.2011.01366.x

[[28]] H Jr Gong, PW Bradley, MS Simmons, DP Tashkin. Impaired exercise performance and pulmonary function in elite cyclists during low-level ozone exposure in a hot environment. Am Rev Respir Dis, 134 (4) ( 1986), pp. 726-733, DOI: 10.1164/arrd.1986.134.4.726

[[29]] WC Adams, ES Schelegle. Ozone and high ventilation effects on pulmonary function and endurance performance. J Appl Physiol Respir Environ Exerc Physiol, 55 (3) ( 1983), pp. 805-812, DOI: 10.1152/jappl.1983.55.3.805

[[30]] LV Giles, MS Koehle. The health effects of exercising in air pollution. Sports Med, 44 (2) ( 2014), pp. 223-249, DOI: 10.1007/s40279-013-0108-z

[[31]] OR Cooper, DD Parrish, J Ziemke, et al.. Global distribution and trends of tropospheric ozone: an observation-based review. Elem Sci Anthropocene, 2 ( 2014), Article 000029, DOI: 10.12952/journal.elementa.000029

[[32]] PS Monks, AT Archibald, A Colette, et al.. Tropospheric ozone and its precursors from the urban to the global scale from air quality to short-lived climate forcer. Atmos Chem Phys, 15 (15) ( 2015), pp. 8889-8973, DOI: 10.5194/ACP-15-8889-2015

[[33]] G Cheng, L Hong, Z Lin, X Yue, R Dang, Y Yang. Persistent ozone pollution episodes in North China exacerbated by regional transport. Environ Pollut, 265 (Pt A) ( 2020), Article 115056, DOI: 10.1016/j.envpol.2020.115056

[[34]] C Braun-Fahrländer, N Künzli, G Domenighetti, CF Carell, U Ackermann-Liebrich. Acute effects of ambient ozone on respiratory function of Swiss schoolchildren after a 10-minute heavy exercise. Pediatr Pulmonol, 17 (3) ( 1994), pp. 169-177, DOI: 10.1002/ppul.1950170306

[[35]] SI Gibbons, WC Adams. Combined effects of ozone exposure and ambient heat on exercising females. J Appl Physiol Respir Environ Exerc Physiol, 57 (2) ( 1984), pp. 450-456, DOI: 10.1152/jappl.1984.57.2.450

[[36]] B Brunekreef, G Hoek, O Breugelmans, M Leentvaar. Respiratory effects of low-level photochemical air pollution in amateur cyclists. Am J Respir Crit Care Med, 150 (4) ( 1994), pp. 962-966, DOI: 10.1164/ajrccm.150.4.7921470

[[37]] L Grievink, SM Jansen, P van’t Veer, B Brunekreef. Acute effects of ozone on pulmonary function of cyclists receiving antioxidant supplements. Occup Environ Med, 55 (1) ( 1998), pp. 13-17, DOI: 10.1136/oem.55.1.13

[[38]] LJ Folinsbee, JF Bedi, SM Horvath. Pulmonary function changes after 1 h continuous heavy exercise in 0.21 ppm ozone. J Appl Physiol Respir Environ Exerc Physiol, 57 (4) ( 1984), pp. 984-988, DOI: 10.1152/jappl.1984.57.4.984

[[39]] TG Bedford, CM Tipton, NC Wilson, RA Oppliger, CV Gisolfi. Maximum oxygen consumption of rats and its changes with various experimental procedures. J Appl Physiol Respir Environ Exerc Physiol, 47 (6) ( 1979), pp. 1278-1283, DOI: 10.1152/jappl.1979.47.6.1278

[[40]] F Qin, Y Dong, S Wang, et al.. Maximum oxygen consumption and quantification of exercise intensity in untrained male Wistar rats. Sci Rep, 10 (1) ( 2020), Article 11520, DOI: 10.1038/s41598-020-68455-8

[[41]] Y Chen, H Li, H Karimian, M Li, Q Fan, Z Xu. Spatio-temporal variation of ozone pollution risk and its influencing factors in China based on Geodetector and Geospatial models. Chemosphere, 302 ( 2022), Article 134843, DOI: 10.1016/j.chemosphere.2022.134843

[[42]] M Hargreaves, LL Spriet. Skeletal muscle energy metabolism during exercise. Nat Metab, 2 (9) ( 2020), pp. 817-828, DOI: 10.1038/s42255-020-0251-4

[[43]] P Moghetti, E Bacchi, C Brangani, S Donà, C Negri. Metabolic effects of exercise. Front Horm Res, 47 ( 2016), pp. 44-57, DOI: 10.1159/000445156

[[44]] F Li, M Xu, M Wang, et al.. Roles of mitochondrial ROS and NLRP 3 inflammasome in multiple ozone-induced lung inflammation and emphysema. Respir Res, 19 (1) ( 2018), p. 230, DOI: 10.1186/s12931-018-0931-8

[[45]] S Kierstein, FR Poulain, Y Cao, et al.. Susceptibility to ozone-induced airway inflammation is associated with decreased levels of surfactant protein D. Respir Res, 7 (1) ( 2006), p. 85, DOI: 10.1186/1465-9921-7-85

[[46]] A Bao, H Yang, J Ji, et al.. Involvements of p 38 MAPK and oxidative stress in the ozone-induced enhancement of AHR and pulmonary inflammation in an allergic asthma model. Respir Res, 18 (1) ( 2017), p. 216, DOI: 10.1186/s12931-017-0697-4

[[47]] VR Sunil, KN Vayas, CB Massa, AJ Cow, JD Laskin, DL Laskin. Ozone-induced injury and oxidative stress in bronchiolar epithelium are associated with altered pulmonary mechanics. Toxicol Sci, 133 (2) ( 2013), pp. 309-319, DOI: 10.1093/toxsci/kft071

[[48]] CH Wiegman, F Li, CJ Clarke, et al.. A comprehensive analysis of oxidative stress in the ozone-induced lung inflammation mouse model. Clin Sci (Lond), 126 (6) ( 2014), pp. 425-440, DOI: 10.1042/CS20130039

[[49]] AP Russell, VC Foletta, RJ Snow, GD Wadley. Skeletal muscle mitochondria: a major player in exercise, health and disease. Biochim Biophys Acta, 1840 (4) ( 2014), pp. 1276-1284, DOI: 10.1016/j.bbagen.2013.11.016

[[50]] M Valdez, JM Valdez, D Freeborn, AFM Johnstone, PRS Kodavanti. The effects of ozone exposure and sedentary lifestyle on neuronal microglia and mitochondrial bioenergetics of female Long-Evans rats. Toxicol Appl Pharmacol, 408 ( 2020), Article 115254, DOI: 10.1016/j.taap.2020.115254

[[51]] GC Chuang, Z Yang, DG Westbrook, et al.. Pulmonary ozone exposure induces vascular dysfunction, mitochondrial damage, and atherogenesis. Am J Physiol Lung Cell Mol Physiol, 297 (2) ( 2009), pp. L209-L216, DOI: 10.1152/ajplung.00102.2009

[[52]] QA Hathaway, N Majumder, WT Goldsmith, et al.. Transcriptomics of single dose and repeated carbon black and ozone inhalation co-exposure highlight progressive pulmonary mitochondrial dysfunction. Part Fibre Toxicol, 18 (1) ( 2021), p. 44, DOI: 10.1186/s12989-021-00437-8

[[53]] MC Valdez, D Freeborn, JM Valdez, et al.. Mitochondrial bioenergetics in brain following ozone exposure in rats maintained on coconut, fish and olive oil-rich diets. Int J Mol Sci, 20 ( 2019), p. 24, DOI: 10.3390/ijms20246303

[[54]] D Tang, R Kang, TV Berghe, P Vandenabeele, G Kroemer. The molecular machinery of regulated cell death. Cell Res, 29 (5) ( 2019), pp. 347-364, DOI: 10.1038/s41422-019-0164-5