Pifu-xingbing zhenliaoxue zazhi (Feb 2023)

Estradiol inhibits phosphorylation of JAK/STAT, PI3K/AKT and expression of IL-17A in mouse mononuclear cells treated with IL-36β

  • Shuang LIU,
  • Shanshan YANG,
  • Chunchan ZHENG,
  • Sujun LUO,
  • Na WEI,
  • Sirui LI,
  • Jianqiang SHI,
  • Rongyi CHEN

DOI
https://doi.org/10.3969/j.issn.1674-8468.2023.01.008
Journal volume & issue
Vol. 30, no. 1
pp. 44 – 49

Abstract

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Objective To investigate the effects of β-estradiol (E2) on the phosphorylation of JAK/STAT, PI3K/AKT signaling pathways and the expression of IL-17A in mouse spleen mononuclear cells treated with IL-36β. Methods The spleens were isolated from female SPF C57BL/6 mice aged 6-8 weeks. The mononuclear cell suspensions were obtained from spleens by density gradient centrifugation. The suspensions of mononuclear cells were transferred into four petri dishes, with 1.5×107 cells/dish, followed by the treatment with either E2 (10-8 mol/L) or IL-36β (10 ng/mL) or IL-36β+E2, while untreated cells served as normal controls. Western blotting was used to assess the changes in phosphorylation of JAK/STAT and PI3K/AKT proteins 30 min after incubation with IL-36β or E2. Moreover, the mononuclear cells were also treated with IL-36β, E2, JAK inhibitor and PI3K inhibitor, respectively, followed by assessment of IL-17A mRNA levels at 6, 12 and 24 h, meanwhile expression levels of IL-17A protein were measured with Western blot at 24 h. Results IL-36β activated the phosphorylation of JAK3/STAT2 and PI3K/AKT in mouse spleen mononuclear cells, while E2 inhibited such activation induced by IL-36β. Expression levels of IL-17A mRNA differed significantly among four groups of cells at 6, 12 and 24 h (F=25.50, 127.04 and 9.76, respectively, all P<0.01), with the highest expression levels in IL-36β-treated cells. Treatment with IL-36β+E2 lowered expression levels of IL-17A mRNA (P<0.05 vs. IL-36β treatment). Moreover, treatment of cells with IL-36β for 24 h increased IL-17A protein expression, while either JAK or PI3K inhibitor attenuated IL-36β-induced upregulation of IL-17A expression. Conclusions IL-36β upregulates IL-17A expression via activation of JAK3/STAT2 and PI3K/AKT signaling pathways in mouse spleen mononuclear cells. E2 inhibits IL-36β-induced activation of JAK3/STAT2 and PI3K/AKT signaling pathways and down-regulates IL-17A expression.

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