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中国应用生理学杂志 ›› 2021, Vol. 37 ›› Issue (5): 449-453.doi: 10.12047/j.cjap.6127.2021.071

• 研究论文 •    下一篇

α1-肾上腺素受体对胶原诱导性关节炎小鼠Treg细胞功能的影响*

王小琴, 石卉, 方小霞, 彭聿平   

  1. 南通大学医学院生理学系, 江苏 南通 226001
  • 收稿日期:2020-07-12 修回日期:2021-02-01 出版日期:2021-09-28 发布日期:2021-11-24
  • 通讯作者: Tel: +86-513-85051714; E-mail: yppeng@ntu.edu.cn
  • 基金资助:
    *国家自然科学基金(31771293);南通大学博士后基金(18B14);南通市科技项目(MS12019011)

Effects of α1-adrenoreceptor on Treg cell function in collagen-induced arthritis

WANG Xiao-qin, SHI Hui, FANG Xiao-xia, PENG Yu-ping   

  1. Department of Physiology, School of Medicine, Nantong University, Nantong 226001, China
  • Received:2020-07-12 Revised:2021-02-01 Online:2021-09-28 Published:2021-11-24

摘要: 目的: 利用胶原诱导性关节炎 (CIA) 模型小鼠,探讨去甲肾上腺素 (NE) 及其α1-肾上腺素受体 (AR ) 对CIA小鼠Treg细胞的作用。方法: 雄性DBA/1小鼠 32 只,随机分为对照组 (n=8) 和CIA模型组 (n=24)。II型胶原 (CII) 乳剂100 μl 尾根部注射DBA/1小鼠制备CIA小鼠模型,在初次免疫后第 41 日,用免疫荧光法检测小鼠脾脏中CD4+T与α1-AR的共定位情况;用Western blot法检测小鼠踝关节和脾脏中α1-AR的蛋白表达。分离纯化CIA小鼠脾脏中CD4+ T细胞,用抗CD3和抗CD28的单克隆抗体刺激CD4+T细胞,进行细胞培养,分为未加药组和加药组,加药组用NE或α1-AR激动剂苯肾上腺素 (phenylephrine) 处理细胞,用流式细胞术检测CIA小鼠CD4+T细胞中Treg的细胞数;用Western blot法检测CIA小鼠CD4+T中转化生长因子-β (TGF-β) 和IL-10的蛋白表达。结果: CD4+ T细胞能够表达α1-AR;与对照组相比,CIA小鼠踝关节和脾脏中α1-AR的蛋白表达显著降低(P<0.01);与未加药的CIA小鼠的CD4+ T细胞相比,NE加入后的CIA小鼠CD4+ T细胞中Treg细胞的功能显著增强(P<0.01);α1-AR激动剂phenylephrine加入后的CIA小鼠CD4+ T细胞中Treg细胞的功能显著增强(P< 0.01)。结论: 激活CIA小鼠CD4+ T细胞上的α1-AR可增强Treg细胞的功能,促进CD4+ T细胞向Treg细胞方向分化,发挥抗炎作用。

关键词: 小鼠, 胶原诱导性关节炎, Treg细胞, 去甲肾上腺素, α1-肾上腺素受体

Abstract: Objective: An animal model of collagen-induced arthritis (CIA) was used to investigate the effects of norepinephrine (NE) and α1-adrenoreceptor (α1-AR) on Treg cells in CIA. Methods: Thirty-two male DBA/1 mice were randomly divided into control group and CIA model group. CIA was prepared by intradermal injection of collagen type II (CII, 100 μl) at the tail base of DBA/1 mice. On the 41th day following primary immunization, co-expression of CD4+T and α1-AR in mouse spleens was observed. Protein expressions of α1-AR in the ankle joints and the spleens of mice were measured by Western blot analysis. The CD4+ T cells were isolated from the mouse spleen tissues in CIA mice and treated with NE or α1-AR agonist phenylephrine. Percentage of Treg cells in mouse CD4+ T cells of CIA mice was determined by flow cytometry. Expressions of α1-AR, transforming growth factor-β (TGF-β) and IL-10 in CD4+T cells of CIA mice were assessed by Western blot. Results: Co-expression of CD4 and α1-AR was observed in spleens of both intact and CIA mice. Compared with intace mice, α1-AR expressions in the ankle joints and spleens were down-regulated in CIA mice. NE increased the function of Treg cells in CD4+ T cells of CIA mice compared with that of nothing-treated CD4+T cells of CIA mice. Moreover, the α1-AR agonist phenylephrine increased the Treg cell function in CD4+ T cells of CIA mice relative to that of nothing-treated CD4+T cells of CIA mice. Conclusion: Activating α1-AR on CD4+T cells of CIA mice enhances Treg cell function,facilitating a shift of CD4+T cells toward Treg polarization.

Key words: mice, collagen-induced arthritis, Treg cells, norepinephrine, α1-adrenoreceptor

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