Do NGF and LPS Interact Synergistically to Modulate Inflammation in Sheep Endometrial Epithelial Cells?
Abstract
1. Introduction
2. Results
2.1. Viability and Epithelial Identity of SELEC
2.2. Prostaglandin Secretion Levels in SELEC Culture Medium
2.3. Gene Expression Signatures in SELECs
3. Discussion
4. Materials and Methods
4.1. Animal Selection and Experimental Design
4.2. Establishment of Primary SELEC Cultures
4.3. Immunofluorescence Characterization of Tight Junctions
4.4. SELEC Incubation with NGF and LPS
4.5. Cell Viability Assay
4.6. Culture Medium: PGE2 and PGF2α Assessment
4.7. SELEC: Gene Expression Signatures
4.8. Statistical Analyses
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- Mravec, B.; Tibensky, M.; Horvathova, L. Stress and Cancer. Part II: Therapeutic Implications for Oncology. J. Neuroimmunol. 2020, 346, 577312. [Google Scholar] [CrossRef] [PubMed]
- Dissen, G.A.; Romero, C.; Hirshfield, A.N.; Ojeda, S.R. Nerve Growth Factor Is Required for Early Follicular Development in the Mammalian Ovary. Endocrinology 2001, 142, 2078–2086. [Google Scholar] [CrossRef] [PubMed]
- Salas, C.; Julio-Pieper, M.; Valladares, M.; Pommer, R.; Vega, M.; Mastronardi, C.; Kerr, B.; Ojeda, S.R.; Lara, H.E.; Romero, C. Nerve Growth Factor-Dependent Activation of trkA Receptors in the Human Ovary Results in Synthesis of Follicle-Stimulating Hormone Receptors and Estrogen Secretion. J. Clin. Endocrinol. Metab. 2006, 91, 2396–2403. [Google Scholar] [CrossRef] [PubMed]
- Minnone, G.; De Benedetti, F.; Bracci-Laudiero, L. NGF and Its Receptors in the Regulation of Inflammatory Response. Int. J. Mol. Sci. 2017, 18, 1028. [Google Scholar] [CrossRef] [PubMed]
- Segal, T.R.; Kim, K.; Mumford, S.L.; Goldfarb, J.M.; Weinerman, R.S. How Much Does the Uterus Matter? Perinatal Outcomes Are Improved When Donor Oocyte Embryos Are Transferred to Gestational Carriers Compared to Intended Parent Recipients. Fertil. Steril. 2018, 110, 888–895. [Google Scholar] [CrossRef] [PubMed]
- Peng, B.; Zhan, H.; Alotaibi, F.; Alkusayer, G.M.; Bedaiwy, M.A.; Yong, P.J. Nerve Growth Factor Is Associated with Sexual Pain in Women with Endometriosis. Reprod. Sci. 2018, 25, 540–549. [Google Scholar] [CrossRef] [PubMed]
- Zhao, B.; Niu, X.; Huang, S.; Yang, J.; Wei, Y.; Wang, X.; Wang, J.; Wang, Y.; Guo, X. TLR4 Agonist and Hypoxia Synergistically Promote the Formation of TLR4/NF-κB/HIF-1α Loop in Human Epithelial Ovarian Cancer. Anal. Cell. Pathol. 2022, 2022, 1–19. [Google Scholar] [CrossRef] [PubMed]
- Machairiotis, N.; Vasilakaki, S.; Thomakos, N. Inflammatory Mediators and Pain in Endometriosis: A Systematic Review. Biomedicines 2021, 9, 54. [Google Scholar] [CrossRef] [PubMed]
- Chen, S.; Liu, Y.; Zhong, Z.; Wei, C.; Liu, Y.; Zhu, X. Peritoneal Immune Microenvironment of Endometriosis: Role and Therapeutic Perspectives. Front. Immunol. 2023, 14, 1134663. [Google Scholar] [CrossRef] [PubMed]
- Chao, M.V. Neurotrophins and Their Receptors: A Convergence Point for Many Signalling Pathways. Nat. Rev. Neurosci. 2003, 4, 299–309. [Google Scholar] [CrossRef] [PubMed]
- Mercati, F.; Guelfi, G.; Martí, J.I.; Dall’Aglio, C.; Calleja, L.; Caivano, D.; Marenzoni, M.L.; Capaccia, C.; Anipchenko, P.; Palermo, F.A.; et al. Seasonal Variation of NGF in Seminal Plasma and Expression of NGF and Its Cognate Receptors NTRK1 and p75NTR in the Sex Organs of Rams. Domest. Anim. Endocrinol. 2024, 89, 106877. [Google Scholar] [CrossRef] [PubMed]
- Maranesi, M.; Boiti, C.; Zerani, M. Nerve Growth Factor (NGF) and Animal Reproduction. In Recent Advances in NGF and Related Molecules: The Continuum of the NGF “Saga”; Calzà, L., Aloe, L., Giardino, L., Eds.; Springer International Publishing: Cham, Switzerland, 2021; pp. 277–287. ISBN 978-3-030-74046-7. [Google Scholar]
- Guelfi, G.; Pasquariello, R.; Dall’Aglio, C.; Mercati, F.; Suvieri, C.; Conte, C.; Capaccia, C.; Ratto, M.; Maranesi, M. The Autocrine Impact of Nerve Growth Factor on Sheep Uterine Epithelial Cells. Cells 2025, 14, 208. [Google Scholar] [CrossRef] [PubMed]
- Herath, S.; Lilly, S.T.; Fischer, D.P.; Williams, E.J.; Dobson, H.; Bryant, C.E.; Sheldon, I.M. Bacterial Lipopolysaccharide Induces an Endocrine Switch from Prostaglandin F2α to Prostaglandin E2 in Bovine Endometrium. Endocrinology 2009, 150, 1912–1920. [Google Scholar] [CrossRef] [PubMed]
- Hubner, A.M.; Canisso, I.F.; Peixoto, P.M.; Coelho, W.M.; Cunha, L.L.; Ribeiro, L.; Crump, S.; Lima, F.S. Effect of Nerve Growth Factor-β Administered at Insemination for Lactating Holstein Dairy Cows Bred after Timed-Artificial Insemination Protocol. J. Dairy Sci. 2022, 105, 6353–6363. [Google Scholar] [CrossRef] [PubMed]
- Tan, H.; Pan, P.; Zhang, L.; Cao, Z.; Liu, B.; Li, H.; Su, X. Nerve Growth Factor Promotes Expression of Costimulatory Molecules and Release of Cytokines in Dendritic Cells Involved in Th2 Response through LPS-Induced p75NTR. J. Asthma 2016, 53, 989–998. [Google Scholar] [CrossRef] [PubMed]
- Jiang, Y.; Chen, G.; Zhang, Y.; Lu, L.; Liu, S.; Cao, X. Nerve Growth Factor Promotes TLR4 Signaling-Induced Maturation of Human Dendritic Cells In Vitro through Inducible p75NTR 1. J. Immunol. 2007, 179, 6297–6304. [Google Scholar] [CrossRef] [PubMed]
- Ferraguti, G.; Fanfarillo, F.; Tarani, L.; Blaconà, G.; Tarani, F.; Barbato, C.; Minni, A.; Ralli, M.; Francati, S.; Greco, A.; et al. NGF and the Male Reproductive System: Potential Clinical Applications in Infertility. Int. J. Mol. Sci. 2022, 23, 13127. [Google Scholar] [CrossRef] [PubMed]
- Stewart, J.L.; Canisso, I.F.; Ellerbrock, R.E.; Mercadante, V.R.G.; Lima, F.S. Nerve Growth Factor-β Production in the Bull: Gene Expression, Immunolocalization, Seminal Plasma Constitution, and Association with Sire Conception Rates. Anim. Reprod. Sci. 2018, 197, 335–342. [Google Scholar] [CrossRef] [PubMed]
- Gajardo, G.; Paiva, L.; Ulloa-Leal, C.; Valderrama, X.; López, G.; Carrasco, A.; Hidalgo, A.I.; Silva, M.E.; Palma, P.I.; Ratto, M.H. Administration of Beta-Nerve Growth Factor during the Preovulatory Stage Improves Endocrine and Luteal Function in Dairy Heifers. Animals 2023, 13, 1004. [Google Scholar] [CrossRef] [PubMed]
- Egashira, M.; Hirota, Y. Uterine Receptivity and Embryo–Uterine Interactions in Embryo Implantation: Lessons from Mice. Reprod. Med. Biol. 2013, 12, 127–132. [Google Scholar] [CrossRef] [PubMed]
- Puspita, R.D.; Rizal, D.M.; Syarif, R.A.; Sari, I.P. Role of COX-2 for Successful Embryo Implantation Process: A Mini-Review. Open Access Maced. J. Med. Sci. 2023, 11, 31–37. [Google Scholar] [CrossRef]
- Duan, T.; Du, Y.; Xing, C.; Wang, H.Y.; Wang, R.-F. Toll-Like Receptor Signaling and Its Role in Cell-Mediated Immunity. Front. Immunol. 2022, 13, 812774. [Google Scholar] [CrossRef] [PubMed]
- Guelfi, G.; Capaccia, C.; Santoro, M.M.; Diverio, S. Identification of Appropriate Endogenous Controls for Circulating miRNA Quantification in Working Dogs under Physiological Stress Conditions. Animals 2023, 13, 576. [Google Scholar] [CrossRef] [PubMed]
- Firmal, P.; Shah, V.K.; Chattopadhyay, S. Insight Into TLR4-Mediated Immunomodulation in Normal Pregnancy and Related Disorders. Front. Immunol. 2020, 11, 807. [Google Scholar] [CrossRef] [PubMed]
- Yan, B.; Yu, X.; Cai, X.; Huang, X.; Xie, B.; Lian, D.; Chen, J.; Li, W.; Lin, Y.; Ye, J.; et al. A Review: The Significance of Toll-Like Receptors 2 and 4, and NF-κB Signaling in Endothelial Cells during Atherosclerosis. Front. Biosci. (Landmark Ed.) 2024, 29, 161. [Google Scholar] [CrossRef] [PubMed]
- Baldassarro, V.A.; Cescatti, M.; Rocco, M.L.; Aloe, L.; Lorenzini, L.; Giardino, L.; Calzà, L. Nerve Growth Factor Promotes Differentiation and Protects the Oligodendrocyte Precursor Cells from in Vitro Hypoxia/Ischemia. Front. Neurosci. 2023, 17, 1111170. [Google Scholar] [CrossRef] [PubMed]
- Schwanhäusser, B.; Busse, D.; Li, N.; Dittmar, G.; Schuchhardt, J.; Wolf, J.; Chen, W.; Selbach, M. Global Quantification of Mammalian Gene Expression Control. Nature 2011, 473, 337–342. [Google Scholar] [CrossRef] [PubMed]
- Marshall, J.S.; Gomi, K.; Blennerhassett, M.G.; Bienenstock, J. Nerve Growth Factor Modifies the Expression of Inflammatory Cytokines by Mast Cells Via a Prostanoid-Dependent Mechanism. J. Immunol. 1999, 162, 4271–4276. [Google Scholar] [CrossRef] [PubMed]
- Heese, K.; Fiebich, B.L.; Bauer, J.; Otten, U. NF-κB Modulates Lipopolysaccharide-Induced Microglial Nerve Growth Factor Expression. Glia 1998, 22, 401–407. [Google Scholar] [CrossRef]
- Grandi, G.; Mueller, M.; Bersinger, N.A.; Cagnacci, A.; Volpe, A.; McKinnon, B. Does Dienogest Influence the Inflammatory Response of Endometriotic Cells? A Systematic Review. Inflamm. Res. 2016, 65, 183–192. [Google Scholar] [CrossRef] [PubMed]
- Imafuku, K.; Iwata, H.; Natsuga, K.; Okumura, M.; Kobayashi, Y.; Kitahata, H.; Kubo, A.; Nagayama, M.; Ujiie, H. Zonula Occludens-1 Distribution and Barrier Functions Are Affected by Epithelial Proliferation and Turnover Rates. Cell Prolif. 2023, 56, e13441. [Google Scholar] [CrossRef] [PubMed]
- Guelfi, G.; Dall’Aglio, C.; Bufalari, A.; Mercati, F.; Anipchenko, P.; Capaccia, C.; Cocci, P.; Palermo, F.A.; Acuti, G.; Troisi, A.; et al. Interleukin-1 Beta (IL1B) and Nerve Growth Factor (NGF): Key Players in Rabbit Reproductive Regulation. Int. J. Mol. Sci. 2024, 25, 10986. [Google Scholar] [CrossRef] [PubMed]
- Livak, K.J.; Schmittgen, T.D. Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2−ΔΔCT Method. Methods 2001, 25, 402–408. [Google Scholar] [CrossRef] [PubMed]
Gene Symbol | TaqMan ID | Sequence ID | Exon | Bp |
---|---|---|---|---|
NGF (nerve growth factor) | Cf02697134_s1 | NM_001194950.1 | 1 | 161 |
NTRK1 (Neurotrophic Receptor Tyrosine Kinase 1) | Oa04767849_g1 | XM_027976575.1 | 14–15 | 114 |
p75NTR (p75 Neurotrophin Receptor) | Oa04853013_m1 | XM_027974687.1 | 2–3 | 62 |
COX2 (Cyclooxygenase 2) | Oa04657348_g1 | NP_001009432.1 | 4–5 | 66 |
STAR (steroidogenic acute regulatory protein) | Oa04657047_m1 | NM_001009243.1 | 4–5 | 69 |
TLR4 (toll-like receptor 4) | Oa04656419_m1 | NM_001135930.1 | 1–2 | 108 |
IGFBP6 (Insulin-Like Growth Factor Binding Protein 6) | Cf02664455_g1 | XP_858473.1 | 3–4 | 105 |
HPRT1 (Hypoxanthine Phosphoribosyltransferase 1) | Oa04825272_gH | XM_015105023.2 | 7–8 | 52 |
TBP (TATA Binding Protein) | Oa04818075_m1 | XM_015097549.2 | 4–5 | 66 |
ACTB (Beta-Actin) | Cf04931159_m1 | NM_001195845.2 | 1 | 52 |
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content. |
© 2025 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Guelfi, G.; Capaccia, C.; Ratto, V.F.; Dall’Aglio, C.; Mercati, F.; Maranesi, M. Do NGF and LPS Interact Synergistically to Modulate Inflammation in Sheep Endometrial Epithelial Cells? Int. J. Mol. Sci. 2025, 26, 6862. https://doi.org/10.3390/ijms26146862
Guelfi G, Capaccia C, Ratto VF, Dall’Aglio C, Mercati F, Maranesi M. Do NGF and LPS Interact Synergistically to Modulate Inflammation in Sheep Endometrial Epithelial Cells? International Journal of Molecular Sciences. 2025; 26(14):6862. https://doi.org/10.3390/ijms26146862
Chicago/Turabian StyleGuelfi, Gabriella, Camilla Capaccia, Vicente Francisco Ratto, Cecilia Dall’Aglio, Francesca Mercati, and Margherita Maranesi. 2025. "Do NGF and LPS Interact Synergistically to Modulate Inflammation in Sheep Endometrial Epithelial Cells?" International Journal of Molecular Sciences 26, no. 14: 6862. https://doi.org/10.3390/ijms26146862
APA StyleGuelfi, G., Capaccia, C., Ratto, V. F., Dall’Aglio, C., Mercati, F., & Maranesi, M. (2025). Do NGF and LPS Interact Synergistically to Modulate Inflammation in Sheep Endometrial Epithelial Cells? International Journal of Molecular Sciences, 26(14), 6862. https://doi.org/10.3390/ijms26146862