Animal Bioscience

Asian Australasian Association of Animal Production Societies (아세아태평양축산학회)
권호 표지
DOI QR코드

DOI QR Code

Distinct damage levels and transcriptional responses of lung in Hezuo pigs and Bama pigs during cold exposure

  • Yajuan Li (College of Animal Science and Technology, Gansu Agricultural University) ;
  • Xiaoli Gao (College of Animal Science and Technology, Gansu Agricultural University) ;
  • Yating Zhang (College of Animal Science and Technology, Gansu Agricultural University) ;
  • Shuangbao Gun (College of Animal Science and Technology, Gansu Agricultural University)
  • Received : 2025.12.01
  • Accepted : 2026.02.27
  • Published : 2026.06.01
Download PDF
⟨ Previous Next ⟩

Abstract

Objective: The aim of the present study is to compare cold adaptation mechanisms between cold-tolerant Hezuo and cold-sensitive Bama pigs. Methods: A total of 40 healthy pigs (75 days old), including 20 Hezuo pigs and 20 Bama pigs, were used in this study. A 2×2 factorial design was employed, with factors including breed (Hezuo vs. Bama) and ambient temperature (23±2℃ vs. -15±2℃). After 7 days of acclimation, pigs in the cold groups were exposed to low temperature and slaughtered on days 0, 1, 5, and 10 (n = 5 per group per time point). All pigs had ad libitum access to feed and water. Lung histology, wet/dry ratio, oxidative and inflammatory biomarkers, apoptosis, and transcriptomics were analyzed. Results: The results showed that Hezuo pigs displayed less severe alveolar septal thickening, inflammatory infiltration, and fine bronchial fold extension during cold exposure than Bama pigs. The W/D ratio dramatically decreased in Hezuo pigs while rising in Bama pigs. Hezuo pigs exhibited significantly higher aquaporin-1 (AQP-1) and aquaporin-5 (AQP-5) expressions than Bama pigs during the middle and late phases. Bama pigs displayed increased reactive oxygen species, malondialdehyde, tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β) and decreased glutathione (GSH) levels. Hezuo pigs maintained stable GSH levels and no significant changes in late-phase inflammatory markers. Bama pigs had a greater apoptosis density and more TUNEL-positive cells than Hezuo pigs, which was related to the down-regulation of B-cell lymphoma 2 (Bcl-2) and the up-regulation of Bcl2-associated X protein (Bax) and Caspase-3. Transcriptomic analysis revealed that in Bama pigs, distinctive genes such as mucin 5B (MUC5B), matrix metallopeptidase 9 (MMP9), alveolar macrophage chemotactic factor-II (AMCF-II), interleukin 22 receptor subunit alpha 1 (IL22RA1), C-C motif chemokine ligand 16 (CCL16), SRY-box transcription factor 9 (SOX9), keratin 5 (KRT5) contribute to mucus hypersecretion, extracellular matrix degradation, and sustained inflammatory chemotaxis, worsening tissue damage. In contrast, Hezuo pigs possess unique genes such as aldehyde dehydrogenase 1 family member A2 (ALDH1A2), acyl-CoA synthetase long chain family member 6 (ACSL6), ACSM5, aldo-keto reductase family 1 member C1 (AKR1C1), nuclear receptor subfamily 4 group A member 3/2 (NR4A3/2), G protein subunit gamma 4 (GNG4), glycogen synthase 2 (GYS2), which enhance lipid metabolism, facilitate aldehyde detoxification, and mitigate oxidative stress, thereby orchestrating a cellular protection. Conclusion: Hezuo pigs exhibit protective molecular mechanisms, suggesting potential targets for cold-resistance breeding.

Keywords

Acknowledgement

Thank Shanghai Peseno Biotechnology Co., Ltd. for performing RNA sequencing analysis. We also appreciate the cooperation of the staff involved in this study.

References

  1. Kim WS, Ghassemi Nejad J, Lee HG. Impact of cold stress on physiological, endocrinological, immunological, metabolic, and behavioral changes of beef cattle at different stages of growth. Animals 2023;13:1073. https://doi.org/10.3390/ani13061073
  2. Sue-Chu M. Winter sports athletes: long-term effects of cold air exposure. Br J Sports Med 2012;46:397-401. https://doi.org/10.1136/bjsports-2011-090822
  3. Valavanidis A, Vlachogianni T, Fiotakis K, Loridas S. Pulmonary oxidative stress, inflammation and cancer: respirable particulate matter, fibrous dusts and ozone as major causes of lung carcinogenesis through reactive oxygen species mechanisms. Int J Environ Res Public Health 2013;10:3886-907. https://doi.org/10.3390/ijerph10093886
  4. Dubey M, Nagarkoti S, Awasthi D, et al. Nitric oxide-mediated apoptosis of neutrophils through caspase-8 and caspase3-dependent mechanism. Cell Death Dis 2016;7:e2348. https://doi.org/10.1038/cddis.2016.248
  5. Liu J, Wu J, Qiao C, et al. Impact of chronic cold exposure on lung inflammation, pyroptosis and oxidative stress in mice. Int Immunopharmacol 2023;115:109590. https://doi.org/10.1016/j.intimp.2022.109590
  6. Sun CY, Zhao YX, Zhong W, et al. The expression of aquaporins 1 and 5 in rat lung after thoracic irradiation. J Radiat Res 2014;55:683-9. https://doi.org/10.1093/jrr/rru008
  7. Teng T, Yang H, Xu T, et al. Activation of inflammatory networks in the lungs caused by chronic cold stress is moderately attenuated by glucose supplementation. Int J Mol Sci 2022; 23:10697. https://doi.org/10.3390/ijms231810697
  8. Luo B, Shi H, Zhang K, et al. Cold stress provokes lung injury in rats co-exposed to fine particulate matter and lipopolysaccharide. Ecotoxicol Environ Saf 2019;168:9-16. https://doi.org/10.1016/j.ecoenv.2018.10.064
  9. Jiao D, Ji K, Wang W, et al. Transcriptome profiles of the liver in two cold-exposed sheep breeds revealed different mechanisms and candidate genes for thermogenesis. Genet Res 2021;2021:5510297. https://doi.org/10.1155/2021/5510297
  10. Hayward L, Robertson CE, McClelland GB. Phenotypic plasticity to chronic cold exposure in two species of Peromyscus from different environments. J Comp Physiol B 2022;192: 335-48. https://doi.org/10.1007/s00360-021-01423-4
  11. Marmol P, Krapacher F, Ibáñez CF. Control of brown adipose tissue adaptation to nutrient stress by the activin receptor ALK7. Elife 2020;9:e54721. https://doi.org/10.7554/eLife.54721
  12. Yang C, Cao C, Liu J, et al. Distinct transcriptional responses of skeletal muscle to short-term cold exposure in Tibetan pigs and Bama pigs. Int J Mol Sci 2023;24:7431. https://doi.org/10.3390/ijms24087431
  13. Ma YF, Han XM, Huang CP, et al. Population genomics analysis revealed origin and high-altitude adaptation of Tibetan pigs. Sci Rep 2019;9:11463. https://doi.org/10.1038/s41598-019-47711-6
  14. He Y, Deng J, Zhou C, et al. Ursodeoxycholic acid alleviates sepsis-induced lung injury by blocking PANoptosis via STING pathway. Int Immunopharmacol 2023;125:111161. https://doi.org/10.1016/j.intimp.2023.111161
  15. Zhao FQ, Zhang ZW, Qu JP, et al. Cold stress induces antioxidants and Hsps in chicken immune organs. Cell Stress Chaperones 2014;19:635-43. https://doi.org/10.1007/s12192-013-0489-9
  16. Joo SY, Park MJ, Kim KH, et al. Cold stress aggravates inflammatory responses in an LPS-induced mouse model of acute lung injury. Int J Biometeorol 2016;60:1217-25. https://doi.org/10.1007/s00484-015-1116-5
  17. Wei H, Zhang Y, Li T, et al. Intermittent mild cold stimulation alleviates cold stress-induced pulmonary fibrosis by inhibiting the TGF-β1/Smad signaling pathway in broilers. Poult Sci 2024;103:103246. https://doi.org/10.1016/j.psj.2023.103246
  18. Yang P, Sjoding MW. Acute respiratory distress syndrome: definition, diagnosis, and routine management. Crit Care Clin 2024;40:309-27. https://doi.org/10.1016/j.ccc.2023.12.003
  19. Nakahira K, Haspel JA, Rathinam VAK, et al. Autophagy proteins regulate innate immune responses by inhibiting the release of mitochondrial DNA mediated by the NALP3 inflammasome. Nat Immunol 2011;12:222-30. https://doi.org/10.1038/ni.1980
  20. Wei H, Li T, Zhang Y, et al. Cold stimulation causes oxidative stress, inflammatory response and apoptosis in broiler heart via regulating Nrf2/HO-1 and NF-κB pathway. J Therm Biol 2023;116:103658. https://doi.org/10.1016/j.jtherbio.2023.103658
  21. Ren J, Long Y, Liu R, Song G, Li Q, Cui Z. Characterization of biological pathways regulating acute cold resistance of zebrafish. Int J Mol Sci 2021;22:3028. https://doi.org/10.3390/ijms22063028
  22. Xu Q, Wang YC, Liu R, et al. Differential gene expression in the peripheral blood of Chinese Sanhe cattle exposed to severe cold stress. Genet Mol Res 2017;16:gmr16029593. https://doi.org/10.4238/gmr16029593
  23. Ji K, Jiao D, Yang G, et al. Transcriptome analysis revealed potential genes involved in thermogenesis in muscle tissue in cold-exposed lambs. Front Genet 2022;13:1017458. https://doi.org/10.3389/fgene.2022.1017458
  24. Cheung SWM, Yiu JHC, Chin KTC, et al. Content of stress granules reveals a sex difference at the early phase of cold exposure in mice. Am J Physiol Endocrinol Metab 2024;326: E29-37. https://doi.org/10.1152/ajpendo.00317.2023
  25. Li W, Chen Y, Zhang Y, et al. Transcriptome analysis revealed potential genes of skeletal muscle thermogenesis in mashen pigs and large white pigs under cold stress. Int J Mol Sci 2023; 24:15534. https://doi.org/10.3390/ijms242115534
  26. Fan X, Cui L, Hou T, Xue X, Zhang S, Wang Z. Stress responses of testicular development, inflammatory and apoptotic activities in male zebrafish (Danio rerio) under starvation. Dev Comp Immunol 2021;114:103833. https://doi.org/10.1016/j.dci.2020.103833
  27. Malone PE, Hernandez MR. 4-Hydroxynonenal, a product of oxidative stress, leads to an antioxidant response in optic nerve head astrocytes. Exp Eye Res 2007;84:444-54. https://doi.org/10.1016/j.exer.2006.10.020
  28. Cao Y, Li J, Qiu S, Ni S, Duan Y. ACSM5 inhibits ligamentum flavum hypertrophy by regulating lipid accumulation mediated by FABP4/PPAR signaling pathway. Biol Direct 2023; 18:75. https://doi.org/10.1186/s13062-023-00436-z
  29. Marszalek JR, Kitidis C, Dirusso CC, Lodish HF. Long-chain acyl-CoA synthetase 6 preferentially promotes DHA metabolism. J Biol Chem 2005;280:10817-26. https://doi.org/10.1074/jbc.M411750200
  30. Ullman JC, Mellem KT, Xi Y, et al. Small-molecule inhibition of glycogen synthase 1 for the treatment of Pompe disease and other glycogen storage disorders. Sci Transl Med 2024; 16:eadf1691. https://doi.org/10.1126/scitranslmed.adf1691
  31. Li L, Zhang H, Min D, et al. Sox9 activation is essential for the recovery of lung function after acute lung injury. Cell Physiol Biochem 2015;37:1113-22. https://doi.org/10.1159/000430236
  32. Kesimer M. Mucins MUC5AC and MUC5B in the airways: MUCing around together. Am J Respir Crit Care Med 2022; 206:1055-7. https://doi.org/10.1164/rccm.202208-1459E
  33. Yu Y, Wang R, Zhang H, Wang J. Circ_0044411 silencing protects infantile pneumonia from lipopolysaccharide-induced cell injury by sponging miR-141-3p to inhibit CCL16 expression. Int Immunopharmacol 2023;114:109425. https://doi.org/10.1016/j.intimp.2022.109425
  34. Zhang Y, Chang M, Xue Q, et al. Intermittent cold stimulation acclimates broilers to acute cold stress by affecting cardiac lipid metabolism. Anim Biosci 2025;38:775-87. https://doi.org/10.5713/ab.24.0389
  35. Cheng S, Shi F, Tang Z, Yang F, Meng Q, Liu T. Identification and characterization of microRNAs in the liver of yak (Bos grunniens) associated with energy deficiency at high-altitude. Anim Biosci 2025;38:1372-83. https://doi.org/10.5713/ab.24.0756
  36. Meenongyai W, Wongpanit K, Khejornsart P, et al. Effects of varying levels of coated cysteamine hydrochloride in diet on growth performance and carcass quality of steers. Anim Biosci 2025;38:2136-48. https://doi.org/10.5713/ab.24.0863