Early Socialization Triggered ROS-Mediated Activation of Canonical NF-κB Pathway Leading to Inflammation of Spleen in Suckling Piglets
Abstract
:1. Introduction
2. Materials and Methods
2.1. Treatment of Experimental Animals
2.2. Behavioral Observation
2.3. Growth Performance and Intake Measurement
2.4. Detection of Reactive Oxygen Species (ROS) and Oxidative Stress Indexes
2.5. Transcriptome Sequencing
2.6. Real-Time Quantitative-PCR Verification
2.7. Western Blot Analysis
2.8. Statistical Analysis
3. Results
3.1. Aggressive Behavior of Piglets
3.2. Growth Performance and Intake Measurement
3.3. Comparison of ROS Expression Levels and Oxidative Stress Indexes
3.4. Transcriptomics DEGs’ Analysis and Validation
3.5. Early Socialization Induced an Abnormally High Expression of NF-κB Signaling Pathway and Inflammation-Related Protein and mRNA Levels in the Spleen of Piglets
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- Morgan, T.; Pluske, J.; Miller, D.; Collins, T.; Barnes, A.L.; Wemelsfelder, F.; Fleming, P.A. Socialising piglets in lactation positively affects their post-weaning behaviour. Appl. Anim. Behav. Sci. 2014, 158, 23–33. [Google Scholar] [CrossRef]
- Peden, R.S.E.; Turner, S.P.; Boyle, L.A.; Camerlink, I. The translation of animal welfare research into practice: The case of mixing aggression between pigs. Appl. Anim. Behav. Sci. 2018, 204, 1–9. [Google Scholar] [CrossRef]
- Alina, S.; Jarkko, K.N.; Mari, H.; Anna, V. Tail biting and production performance in fattening pigs. Livest. Sci. 2012, 143, 220–225. [Google Scholar] [CrossRef]
- D’Eath, R.B. Socialising piglets before weaning improves social hierarchy formation when pigs are mixed post-weaning. Appl. Anim. Behav. Sci. 2005, 93, 199–211. [Google Scholar] [CrossRef]
- Salazar, L.C.; Ko, H.L.; Yang, C.H.; Llonch, L.; Manteca, X.; Camerlink, I.; Llonch, P. Early socialization as a strategy to increase piglets’social skills in intensive farming conditions. Appl. Anim. Behav. Sci. 2018, 206, 25–31. [Google Scholar] [CrossRef]
- Ko, H.L.; Chong, Q.; Escribano, D.; Camerlink, I.; Manteca, X.; Llonch, P. Pre-weaning socialization and environmental enrichment affect life-long response to regrouping in commercially-reared pigs. Appl. Anim. Behav. Sci. 2020, 229, 105044. [Google Scholar] [CrossRef]
- Becker, B.A.; Misfeldt, M.L. Evaluation of the mitogen-induced proliferation and cell surface differentiation antigens of lymphocytes from pigs 1 to 30 days of age. J. Anim. Sci. 1993, 71, 2073–2078. [Google Scholar] [CrossRef]
- Ji, W.B.; Li, X.; Zhang, X.H.; Liu, R.Z.; Shu, Y.F.; Cheng, Z.; Bi, Y.J.; Liu, H.G.; Bao, J. Effects of two different early socialization models on social behavior and physiology of suckling piglets. Appl. Anim. Behav. Sci. 2021, 243, 105436. [Google Scholar] [CrossRef]
- Camerlink, I.; Farish, M.; D’Eath, R.B.; Arnott, G.; Turner, S.P. Long term benefits on social behaviour after early life socialization of piglets. Animals 2018, 8, 192. [Google Scholar] [CrossRef]
- Bronte, V.; Pittet, M.J. The Spleen in Local and Systemic Regulation of Immunity. Immunity 2013, 39, 806–818. [Google Scholar] [CrossRef]
- Hayden, M.S.; Ghosh, S. Shared Principles in NF-κB Signalling. Cell 2008, 132, 344–362. [Google Scholar] [CrossRef] [PubMed]
- Hayden, M.S.; Ghosh, S. NF-κB in immunobiology. Cell Res. 2011, 21, 223–244. [Google Scholar] [CrossRef]
- Klement, J.F.; Rice, N.R.; Car, B.D.; Abbondanzo, S.J.; Powers, G.D.; Bhatt, P.H.; Chen, C.H.; Rosen, C.A.; Stewart, C.L. I-kappaB-alpha deficiency results in a sustained NF-kappaB response and severe widespread dermatitis in mice. Mol. Cell. Biol. 1996, 16, 2341–2349. [Google Scholar] [CrossRef] [PubMed]
- Beaulieu, M.; Mboumba, S.; Willaume, E.; Kappeler, P.M.; Charpentier, M.J.E. The oxidative cost of unstable social dominance. J. Exp. Biol. 2014, 217, 2629–2632. [Google Scholar] [CrossRef]
- Felippe, R.M.; Oliveira, G.M.; Barbosa, R.S.; Esteves, B.D.; Gonzaga, B.M.S.; Horita, S.I.M.; Garzoni, L.R.; Beghini, D.G.; Araujo-Jorge, T.C.; Fragoso, V.M.S. Experimental Social Stress: Dopaminergic Receptors, Oxidative Stress, and c-Fos Protein Are Involved in Highly Aggressive Behavior. Front. Cell. Neurosci. 2021, 15, 696834. [Google Scholar] [CrossRef] [PubMed]
- Zhang, S.Q.; Kovalenko, A.; Cantarella, G.; Wallach, D. Recruitment of the IKK signalosome to the p55 TNF receptor: RIP and A20 bind to NEMO (IKKgamma) upon receptor stimulation. Immunity 2000, 12, 301–311. [Google Scholar] [CrossRef]
- Liu, H.G.; Wang, C.; Lv, J.; Yu, Y.Q.; Zhang, J.L.; Bao, J. Behavior and performance in primiparous sows of two Min pig hybrid breeds in outdoor housing systems. Appl. Anim. Behav. Sci. 2013, 146, 37–44. [Google Scholar] [CrossRef]
- Altmann, J. Observational Study of Behavior: Sampling Methods. Behaviour 1974, 49, 227–266. [Google Scholar] [CrossRef]
- Kelley, K.W.; McGlone, J.J.; Gaskins, C.T. Porcine aggression: Measurement and effects of crowding and fasting. J. Anim. Sci. 1980, 50, 336–341. [Google Scholar] [CrossRef]
- Jensen, P. Effects of confinement on social interaction patterns in dry sows. Appl. Anim. Behav. Sci. 1984, 12, 93–101. [Google Scholar] [CrossRef]
- Worobec, E.K.; Duncan, I.J.H.; Widowski, T. The effects of weaning at 7, 14 and 28 days on piglet behaviour. Appl. Anim. Behav. Sci. 1999, 62, 173–182. [Google Scholar] [CrossRef]
- Morrison, R.S.; Hemsworth, P.H.; Cronin, G.M.; Campbell, R.G. The social and feeding behaviour of growing pigs in deep-litter, large group housing systems. Appl. Anim. Behav. Sci. 2003, 82, 173–188. [Google Scholar] [CrossRef]
- Lucas, M.E.; Hemsworth, L.M.; Hemsworth, P.H. Review: Early life piglet experiences and impacts on immediate and longer-term adaptability. Animal 2023, 100889. [Google Scholar] [CrossRef] [PubMed]
- Sturza, A.; Popoiu, C.M.; Ionică, M.; Duicu, O.M.; Olariu, S.; Muntean, D.M.; Boia, E.S. Monoamine Oxidase-Related Vascular Oxidative Stress in Diseases Associated with Inflammatory Burden. Oxidative Med. Cell. Longev. 2019, 2019, 8954201. [Google Scholar] [CrossRef] [PubMed]
- Rammal, H.; Bouayed, J.; Soulimani, R. A direct relationship between aggressive behavior in the resident/intruder test and cell oxidative status in adult male mice. Eur. J. Pharmacol. 2010, 627, 173–176. [Google Scholar] [CrossRef] [PubMed]
- Gloire, G.; Legrand-Poels, S.; Piette, J. NF-κB activation by reactive oxygen species: Fifteen years later. Biochem. Pharmacol. 2006, 72, 1493–1505. [Google Scholar] [CrossRef] [PubMed]
- Gilmore, T.D. Introduction to NF-κB: Players, pathways, perspectives. Oncogene 2006, 25, 6680–6684. [Google Scholar] [CrossRef] [PubMed]
- Chen, L.F.; Greene, W.C. Shaping the nuclear action of NF-kappaB. Nat. Rev. Mol. Cell Biol. 2004, 5, 392–401. [Google Scholar] [CrossRef]
- Blonska, M.; Shambharkar, P.B.; Kobayashi, M.; Zhang, D.; Sakurai, H.; Su, B.; Lin, X. TAK1 is recruited to the tumor necrosis factor-alpha (TNF-alpha) receptor 1 complex in a receptor-interacting protein (RIP)-dependent manner and cooperates with MEKK3 leading to NF-kappaB activation. J. Biol. Chem. 2005, 280, 43056–43063. [Google Scholar] [CrossRef]
- Simeonova, P.P.; Toriumi, W.; Kommineni, C.; Erkan, M.; Munson, A.E.; Rom, W.N.; Luster, M.I. Molecular regulation of IL-6 activation by asbestos in lung epithelial cells: Role of reactive oxygen species. J. Immunol. 1997, 159, 3921–3928. [Google Scholar] [CrossRef]
- Bonizzi, G.; Piette, J.; Merville, M.P.; Bours, V. Cell type-specific role for reactive oxygen species in nuclear factor-kappaB activation by interleukin-1. Biochem. Pharmacol. 2000, 59, 7–11. [Google Scholar] [CrossRef] [PubMed]
- Liu, J.; Wang, S.; Zhang, Q.; Li, X.; Xu, S. Selenomethionine alleviates LPS-induced chicken myocardial inflammation by regulating the miR-128-3p-p38 MAPK axis and oxidative stress. Metallomics 2019, 12, 54–64. [Google Scholar] [CrossRef] [PubMed]
- Van Noort, J. Stress proteins in CNS inflammation. J. Pathol. 2008, 214, 267–275. [Google Scholar] [CrossRef] [PubMed]
Item (%) | Creep Feed | Sow Compound Feed |
---|---|---|
Crude protein | ≥18.00 | ≥17.00 |
Crude fiber | ≤4.00 | ≤7.00 |
Crude ash | ≤7.00 | ≤9.00 |
Calcium | 0.70–1.50 | 0.70–1.30 |
Total phosphorus | ≥0.50 | ≥0.50 |
Sodium chloride | 0.30–1.50 | 0.30–1.20 |
Water | ≤14.00 | ≤14.00 |
Lysine | ≥1.35 | ≥1.10 |
Aggressive Behavior | Description |
---|---|
Fighting | The pigs spontaneously and violently bit the head, ear, and neck of the other pig, with tension of the body and tail, which lasted for more than 3 s. [19] |
Head knocking | A pig pushed the head, neck, or body of another pig quickly with its head, touching each other but not biting [20]. |
Gene | Forward Primer (5′→3′) | Reverse Primer (5′→3′) |
---|---|---|
β-actin | GGCACCACACCTTCTACAACGAG | TCATCTTCTCACGGTTGGCTTTGG |
p65 | ACAACCCCTTCCAAGTTCCC | GCACGGTTGTCAAAGATGGG |
IKK-β | AGAGTTCTGCTCGGTCCCTTGTAG | CTTGAGGAGTTACCACGCATGACAG |
IkB-α | TGGTGTCGCTCTTGTTGAAGTGTG | GCTGCTGTATCCGAGTGCTTGG |
IkB-β | CTGCCTGTCCAAGATGAAGAACTCC | GTCCGATGTGATCCCAAACTCTGTC |
IL-1β | GCCAGTCTTCATTGTTCAGGTT | GGAGGGATTCTTCATCGGCTT |
IL-6 | CTGGGGCACCTGAGATTGATG | ACAACAAGGATCTTCAAGCCGT |
TNF-α | CCTCACTCACACCATCAGCC | TCCAGATAGTCGGGCAGGTT |
iNOS | TGGGTTGAATCTGGGTGAAGAG | TCTTGGAAAGTCATCCCGCT |
COX2 | TCCTGAACACCTCCGCTTTG | CATTCCGGGTGCTCCTGTTT |
HSP27 | GGCGTCTCGGAGATCCAGCAG | TGCTCATCCTGCCGCTCCTC |
HSP40 | CATTCAGCCAGATGTCGCAGGAG | GAGTACCGTGTGATGAGCCAGTTG |
HSP60 | TTCGTCAGATTAGGCCGGTG | TTTGGCCCCATAGTAACGGC |
HSP90 | ATCGCCCAGTTGATGTCGTT | TATCGTGAGGGTCCGGTCTT |
PKA | CTGCGTCTGGTCCATTGCTAGTG | TTCTGATGGGCACCTGGAGAGAG |
KCNMB4 | CTGCTGTAGCAGAAACCCCT | TGAAACAGACCTGCCCTTCC |
ATP6V1B1 | CAACGTGCTCCCTTCTCTGT | CAGGTCCTCGGAAGTAAGCG |
HEPACAM | TGGGAAAAGGCAGAACTGGG | GTCTGCTGGGGTCCTACCTT |
NME3 | CGCCTGGTGAAGTACATGGG | CCTCGATGCAGAAATCGCC |
HS3ST3B1 | GCATCATCAGCGACAAGCAC | TAATAAGCCCTGGGCGAAGTC |
SVIP | ACAACGCAGTTTGCAAGGC | ACATCCAAAATGCCCCGAGA |
FAM209B | TCTCAGTAGAGTCGCAGGGT | GGCTCATGGTGGGTTTGTCT |
FOXH1 | GAGTGCCAGTGGAAGAGAGG | TCTGAGGGAAGAGACGAGGG |
FCN2 | GCCTCAAATTGTGCAGTGCT | TAGGTGGCCCGAAACTTCATC |
Aggressive Behavior | CON | IC | SEp | t Value | p Value |
---|---|---|---|---|---|
Fighting | 4.14 | 7.39 * | 0.59 | 6.838 | 0.002 |
Head knocking | 2.86 | 4.78 * | 0.20 | 9.449 | 0.001 |
Item | IC | CON | SEp | t Value | p Vaule |
---|---|---|---|---|---|
Birth weight | 1.46 | 1.44 | 0.13 | 0.377 | 0.710 |
Weaning weight | 10.47 | 10.22 | 0.38 | 2.047 | 0.096 |
Feed consumption | 0.063 | 0.057 | 0.006 | 1.435 | 0.211 |
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Yang, Y.; Wu, M.; Zhang, X.; Zhao, Y.; Zhou, S.; Ji, W.; Liu, H. Early Socialization Triggered ROS-Mediated Activation of Canonical NF-κB Pathway Leading to Inflammation of Spleen in Suckling Piglets. Agriculture 2024, 14, 992. https://doi.org/10.3390/agriculture14070992
Yang Y, Wu M, Zhang X, Zhao Y, Zhou S, Ji W, Liu H. Early Socialization Triggered ROS-Mediated Activation of Canonical NF-κB Pathway Leading to Inflammation of Spleen in Suckling Piglets. Agriculture. 2024; 14(7):992. https://doi.org/10.3390/agriculture14070992
Chicago/Turabian StyleYang, Yue, Mengyao Wu, Xiaolong Zhang, Yunlong Zhao, Sitong Zhou, Wenbo Ji, and Honggui Liu. 2024. "Early Socialization Triggered ROS-Mediated Activation of Canonical NF-κB Pathway Leading to Inflammation of Spleen in Suckling Piglets" Agriculture 14, no. 7: 992. https://doi.org/10.3390/agriculture14070992