ORIGINAL PAPER
 
KEYWORDS
TOPICS
ABSTRACT
Normal pregnancy is characterised by a systemic immunological tolerance against foetal antigens, and the spleen contributes to the adaptive immune tolerance during pregnancy. Nuclear factor kappa B (NF-κB) signallings participate in splenic immune regulation, but it is unclear whether there are changes in NF-κB components expression in the ovine spleen during early pregnancy. The objective of this study was to explore the effects of early pregnancy on the expression of NF-κB components in the maternal spleen in sheep. The spleens were sampled on day 16 of the oestrous cycle, and on days 13, 16 and 25 of gestation. The expression of NF-κB components, including NF-κB1 (p50), NF-κB2 (p52), RelA (p65), RelB and c-Rel, were detected by quantitative real-time PCR, Western blot analysis and immunohistochemical analysis. The results showed that NF-κB1 and RelB mRNA and proteins decreased at days 13 and 16 of pregnancy, but increased at day 25 of pregnancy in comparison with that on day 16 of the oestrous cycle. Nevertheless, NF-κB2 and RelA mRNA and proteins peaked at days 13 and 16 of pregnancy. In addition, early pregnancy inhibited C-Rel expression at days 16 to 25 of pregnancy in the maternal spleen. In conclusion, the variable expression of individual NF-κB components was found in the ovine spleen during early pregnancy, which may be related with pregnancy recognition, and essential for the embryo implantation and pregnancy maintenance.
ACKNOWLEDGEMENTS
The current study was supported by the grants from Natural Science Foundation of Hebei Province, China (C2021402019), and Hebei Science and Technology Bureau, China (21326601D).
CONFLICT OF INTEREST
The authors declare that there is no conflict of interest.
METADATA IN OTHER LANGUAGES:
Chinese
早期妊娠对绵羊脾脏表达核因子-κB组分的影响 摘要
核因子-κB组分, 妊娠, 绵羊, 脾脏
正常妊娠情况下,母体对胎儿抗原产生系统性免疫耐受。在妊娠期间,脾脏参与母体适应性免疫耐受 的形成。核因子-κB (Nuclear factor kappa B, NF-κB) 信号参与脾脏的免疫调节,但是,妊娠早期绵羊脾脏表达 NF-κB组分是否发生改变,目前还不清楚。本研究的目的是探讨早期妊娠对绵羊母体脾脏表达NF-κB组分的 影响。采集发情周期第16天,以及妊娠第13、16和25天的母羊脾脏,采用实时定量PCR、蛋白印迹和免疫 组化分析NF-κB组分的表达,NF-κB组分包括NF-κB1 (p50)、NF-κB2 (p52)、RelA (p65)、RelB和C-Rel。试验结 果表明,与发情周期第16天相比,在妊娠第13天和16天,脾脏表达NF-κB1和 RelB的mRNA和蛋白质减少, 但是,在妊娠第25天却增加。此外,在妊娠第13天和16天,NF-κB2和RelA的mRNA和蛋白质水平达到峰值。 同时,从妊娠第16至25天,母体脾脏C-Rel的表达水平逐步降低。综上所述,在绵羊妊娠早期,脾脏表达 NF-κB不同组分发生改变,这可能与妊娠识别有关,对胚胎附植和妊娠维持至关重要。
 
REFERENCES (41)
1.
Bai J., Zhang L., Zhao Z., Li N., Wang B., Yang L., 2020. Expression of melatonin receptors and CD4 in the ovine thymus, lymph node, spleen and liver during early pregnancy. Immunology 160, 52–63, https://doi.org/10.1111/imm.13....
 
2.
Basavarajappa S.C., Ramakrishnan P., 2020. Regulation of B-cell function by NF-kappaB c-Rel in health and disease. Cell Mol. Life Sci. 77, 3325–3340, https://doi.org/10.1007/s00018....
 
3.
Baud V., Collares D., 2016. Post-translational modifications of RelB NF-κB subunit and associated functions. Cells 5, 22, https://doi.org/10.3390/cells5....
 
4.
Beinke S., Ley S.C., 2004. Functions of NF-kappaB1 and NF-kappaB2 in immune cell biology. Biochem. J. 382, 393–409, https://doi.org/10.1042/BJ2004....
 
5.
Bozkurt M., Şahin L., Ulaş M., 2015. Hysteroscopic polypectomy decreases NF-κB1 expression in the mid-secretory endometrium of women with endometrial polyp. Eur. J. Obstet. Gynecol. Reprod. Biol. 189, 96–100, https://doi.org/10.1016/j.ejog....
 
6.
Buska-Mach K., Kedzierska A.E., Lepczynski A. et al., 2021. Differential signals from TNFα-treated and untreated embryos in uterine tissues and splenic CD4+ T lymphocytes during preimplantation pregnancy in mice. Front. Vet. Sci. 8, 641553, https://doi.org/10.3389/fvets.....
 
7.
Caamaño J.H., Rizzo C.A., Durham S.K., Barton D.S., Raventós-Suárez C., Snapper C.M., Bravo R., 1998. Nuclear factor (NF)-kappa B2 (p100/p52) is required for normal splenic microarchitecture and B cell-mediated immune responses. J. Exp. Med. 187, 185–196, https://doi.org/10.1084/jem.18....
 
8.
Cao N., Cao L., Gao M., Wang H., Zhang L., Yang L., 2021. Changes in mRNA and protein levels of gonadotropin releasing hormone and receptor in ovine thymus, lymph node, spleen, and liver during early pregnancy. Domest. Anim. Endocrinol. 76, 106607, https://doi.org/10.1016/j.doma....
 
9.
Chapman N.R., Europe-Finner G.N., Robson S.C., 2004. Expression and deoxyribonucleic acid-binding activity of the nuclear factor kappaB family in the human myometrium during pregnancy and labor. J. Clin. Endocrinol. Metab. 89, 5683–5693, https://doi.org/10.1210/jc.200....
 
10.
Dai R., Phillips R.A., Ahmed S.A., 2007. Despite inhibition of nuclear localization of NF-kappa B p65, c-Rel, and RelB, 17-beta estradiol up-regulates NF-kappa B signaling in mouse splenocytes: the potential role of Bcl-3. J. Immunol. 179, 1776–1783, https://doi.org/10.4049/jimmun....
 
11.
Fuhler G.M., 2020. The immune system and microbiome in pregnancy. Best Pract. Res. Clin. Gastroenterol. 44–45, 101671, https://doi.org/10.1016/j.bpg.....
 
12.
Godkin J.D., Bazer F.W., Moffatt J., Sessions F., Roberts R.M., 1982. Purification and properties of a major, low molecular weight protein released by the trophoblast of sheep blastocysts at day 13–21. J. Reprod. Fertil. 65, 141–150, https://doi.org/10.1530/jrf.0.....
 
13.
Hamon M.H., Heap R.B., 1990. Progesterone and oestrogen concentrations in plasma of Barbary sheep (aoudad, Ammotragus lervia) compared with those of domestic sheep and goats during pregnancy. J. Reprod. Fertil. 90, 207–211, https://doi.org/10.1530/jrf.0.....
 
14.
Heilmann R.M., Allenspach K., 2017. Pattern-recognition receptors: signaling pathways and dysregulation in canine chronic enteropathies-brief review. J. Vet. Diagn. Invest. 29, 781–787, https://doi.org/10.1177/104063....
 
15.
Kandil D., Leiman G., Allegretta M., Trotman W., Pantanowitz L., Goulart R., Evans M., 2007. Glypican-3 immunocytochemistry in liver fine-needle aspirates: a novel stain to assist in the differentiation of benign and malignant liver lesions. Cancer 111, 316–322, https://doi.org/10.1002/cncr.2....
 
16.
Kashimura M., 2020. The human spleen as the center of the blood defense system. Int. J. Hematol. 112, 147–158, https://doi.org/10.1007/s12185....
 
17.
Lee J., Banu S.K., McCracken J.A., Arosh J.A., 2016. Early pregnancy modulates survival and apoptosis pathways in the corpus luteum in sheep. Reproduction 151, 187–202, https://doi.org/10.1530/REP-15....
 
18.
Li N., Zhao Z., Bai J., Liu B., Mi H., Zhang L., Li G., Yang L., 2019. Characterization of the Th cytokines profile in ovine spleen during early pregnancy. J. Appl. Anim. Res. 47, 386–393,https://doi.org/10.1080/097121....
 
19.
Livak K.J., Schmittgen T.D., 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 25, 402–408, https://doi.org/10.1006/meth.2....
 
20.
Masat E., Gasparini C., Agostinis C., Bossi F., Radillo O., De Seta F., Tamassia N., Cassatella M.A., Bulla R., 2015. RelB activation in anti-inflammatory decidual endothelial cells: a master plan to avoid pregnancy failure? Sci. Rep. 5, 14847, https://doi.org/10.1038/srep14....
 
21.
McNatty K.P., Revefeim K.J., Young A., 1973. Peripheral plasma progesterone concentrations in sheep during the oestrous cycle. J. Endocrinol. 58, 219–225, https://doi.org/10.1677/joe.0.....
 
22.
Nakada D., Oguro H., Levi B.P., Ryan N., Kitano A., Saitoh Y., Takeichi M., Wendt G.R., Morrison S.J., 2014. Oestrogen increases haematopoietic stem-cell self-renewal in females and during pregnancy. Nature 505, 555–558, https://doi.org/10.1038/nature....
 
23.
Nakamura H., Kimura T., Ogita K., Nakamura T., Takemura M., Shimoya K., Koyama S., Tsujie T., Koyama M., Murata Y.,Y., 2004. NF-kappaB activation at implantation window of the mouse uterus. Am. J. Reprod. Immunol. 51, 16–21, https://doi.org/10.1046/j.8755....
 
24.
Norton M.T., Fortner K.A., Bizargity P., Bonney E.A., 2009. Pregnancy alters the proliferation and apoptosis of mouse splenic erythroid lineage cells and leukocytes. Biol. Reprod. 81, 457–464, https://doi.org/10.1095/biolre....
 
25.
Oh S.Y., Hwang J.R., Choi M., Kim Y.M., Kim J.S., Suh Y.L., Choi S.J., Roh C.R., 2020. Autophagy regulates trophoblast invasion by targeting NF-κB activity. Sci. Rep. 10, 14033, https://doi.org/10.1038/s41598....
 
26.
Olmos-Ortiz A., Déciga-García M., Preciado-Martínez E. et al., 2019. Prolactin decreases LPS-induced inflammatory cytokines by inhibiting TLR-4/NFκB signaling in the human placenta. Mol. Hum. Reprod. 25, 660–667, https://doi.org/10.1093/molehr....
 
27.
Ott T.L., 2020. Immunological detection of pregnancy: Evidence for systemic immune modulation during early pregnancy in ruminants. Theriogenology 150, 498–503, https://doi.org/10.1016/j.ther....
 
28.
Patel H., Zaghloul N., Lin K., Liu S.F., Miller E.J., Ahmed M., 2017. Hypoxia-induced activation of specific members of the NF-kB family and its relevance to pulmonary vascular remodeling. Int.J. Biochem. Cell Biol. 92, 141–147, https://doi.org/10.1016/j.bioc....
 
29.
Prusty B.K., Hedau S., Singh A., Kar P., Das B.C., 2007. Selective suppression of NF-kBp65 in hepatitis virus-infected pregnant women manifesting severe liver damage and high mortality. Mol. Med. 13, 518–526, https://doi.org/10.2119/2007-0....
 
30.
Rocha C.C., da Silveira J.C., Forde N., Binelli M., Pugliesi G., 2021. Conceptus-modulated innate immune function during early pregnancy in ruminants: a review. Anim. Reprod. 18, e20200048, https://doi.org/10.1590/1984-3....
 
31.
Ross J.W., Ashworth M.D., Mathew D., Reagan P., Ritchey J.W., Hayashi K., Spencer T.E., Lucy M., Geisert R.D., 2010. Activation of the transcription factor, nuclear factor kappa-B, during the estrous cycle and early pregnancy in the pig. Reprod. Biol.Endocrinol. 8, 39, https://doi.org/10.1186/1477-7....
 
32.
Saha I., Jaiswal H., Mishra R. et al., 2020. RelB suppresses type I Interferon signaling in dendritic cells. Cell Immunol. 349, 104043, https://doi.org/10.1016/j.cell....
 
33.
Sekiya Y., Yamamoto E., Niimi K., Nishino K., Nakamura K., Kotani T., Kajiyama H., Shibata K., Kikkawa F., 2017. c-Rel promotes invasion of choriocarcinoma cells via PI3K/AKT signaling. Oncology 92, 299–310, https://doi.org/10.1159/000458....
 
34.
Shukla V., Kaushal J.B., Sankhwar P., Manohar M., Dwivedi A., 2019. Inhibition of TPPP3 attenuates β-catenin/NF-κB/COX-2 signaling in endometrial stromal cells and impairs decidualization. J. Endocrinol. 240, 417–429, https://doi.org/10.1530/JOE-18....
 
35.
Spencer T.E., Forde N., Lonergan P., 2016. The role of progesterone and conceptus-derived factors in uterine biology during early pregnancy in ruminants. J. Dairy Sci. 99, 5941–5950, https://doi.org/10.3168/jds.20....
 
36.
Wang Y., Han X., Zhang L., Cao N., Cao L., Yang L., 2019. Early pregnancy induces expression of STAT1, OAS1 and CXCL10 in ovine spleen. Animals 9, 882, https://doi.org/10.3390/ani911....
 
37.
Wirasinha R.C., Davies A.R., Srivastava M. et al., 2021. Nfkb2 variants reveal a p100-degradation threshold that defines autoimmune susceptibility. J. Exp. Med. 218, e20200476, https://doi.org/10.1084/jem.20....
 
38.
Xue P., Fan W., Diao Z. et al., 2020. Up-regulation of PTEN via LPS/AP-1/NF-κB pathway inhibits trophoblast invasion contributing to preeclampsia. Mol. Immunol. 118, 182–190, https://doi.org/10.1016/j.moli....
 
39.
Yamazaki T., Kurosaki T., 2003. Contribution of BCAP to maintenance of mature B cells through c-Rel. Nat. Immunol. 4, 780– 86,https://doi.org/10.1038/ni949.
 
40.
Yang L., Guo R., Yao X., Yan J., Bai Y., Zhang L., 2018. Expression of progesterone receptor and progesterone-induced blocking factor in the spleen during early pregnancy in ewes. Livest. Sci. 209, 14–19, https://doi.org/10.1016/j.livs....
 
41.
Yang L., Liu Y., Lv W., Wang P., Wang B., Xue J., Zhang L., 2018. Expression of interferon-stimulated gene 15-kDa protein, cyclooxygenase (COX) 1, COX-2, aldo-keto reductase family 1, member B1, and prostaglandin E synthase in the spleen during early pregnancy in sheep. Anim. Sci. J. 89, 1540–1548, https://doi.org/10.1111/asj.13....
 
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