Effects of Spermidine on Cell Proliferation, Migration, and Inflammatory Response in Porcine Enterocytes

¹ Laboratory of Feed Biotechnology of Ministry of Agriculture and Rural Affairs, Institute of Feed Research, Chinese Academy of Agricultural Sciences, 100081 Beijing, China

^*Correspondence: lixilong@caas.cn (Xi-long Li)
^†These authors contributed equally.
Academic Editor: Guoyao Wu

Front. Biosci. (Landmark Ed) 2022, 27(6), 194; https://doi.org/10.31083/j.fbl2706194

Submitted: 26 April 2022 | Revised: 26 May 2022 | Accepted: 8 June 2022 | Published: 15 June 2022

(This article belongs to the Special Issue Nutrition, Metabolism, and Functions of Amino Acids and Peptides in Animals)

This is an open access article under the CC BY 4.0 license.

Abstract

Background: Polyamines have been demonstrated to be beneficial to porcine intestinal development. Our previous study showed that putrescine mitigates intestinal atrophy in weanling piglets and suppresses inflammatory response in porcine intestinal epithelial cells, it is still unknown the role of spermidine in mediating putrescine function. Objective: The current study aimed to investigate the effect of spermidine on the proliferation, migration, and inflammatory response in porcine intestinal epithelial cells (IPEC-J2 cell line). Methods: The effects of spermidine on proliferation and migration of IPEC-J2 cells were measured. Difluoromethyl ornithine (DFMO) and diethylglyoxal bis (guanylhydrazone) (DEGBG) were used to block the production of putrescine and spermidine, respectively. A cell inflammation model was established with lipopolysaccharides (LPS) stimulation. Gene expression and protein abundance were determined by real-time quantitative PCR and western blotting, respectively. Result: Spermidine significantly enhanced cell proliferation in DFMO (or/and) DEGBG treated IPEC-J2 cells (p $<$ 0.05). Pretreatment with putrescine restored cell growth inhibited by DFMO but did not prevent the decrease in cell proliferation caused by DEGBG (p $>$ 0.05). Similarly, spermidine but not putrescine significantly elevated the rate of migration in DEGBG treated IPEC-J2 cells (p $<$ 0.05). Spermidine deprivation by DEGBG dramatically enhanced mRNA abundance of pro-inflammatory cytokines IL-8, IL-6, and TNF- $\alpha{}$ (p $<$ 0.05), and the addition of spermidine attenuated excessive expression of those inflammatory pro-inflammatory cytokines, moreover, spermidine but not putrescine suppressed the phosphorylation of NF- $\kappa{}$ B induced by DEGBG. Spermidine supplementation also significantly suppressed LPS-induced the expression of TNF- $\alpha{}$ . Conclusions: The present study highlights a novel insight that putrescine may be converted into spermidine to modulate cell proliferation, migration, and inflammatory response on porcine enterocytes.

Keywords

spermidine

putrescine

cell proliferation

cell migration

inflammatory response

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Fig. 1.

Effects of different spermidine concentrations on the growth of IPEC-J2 cells. IPEC-J2 cells were seeded into 96-well plates at a number of 0.5 $\times{}$ 10 ${}^{5}$ cells per well and then treated with 0, 1, 2, 4, 8, 16, 32, 64 $\mu{}$ mol/L spermidine. Cell number was detected at 24 h, 48 h, 72 h, and 96 h with CCK-8 reagent. The data were expressed as the mean $\pm{}$ SE, n = 6. Different letters indicate a significant difference among treatments at each time point (p $<$ 0.05).

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