上海交通大学学报（医学版）

上海交通大学学报（医学版）

• 论著（基础研究） • 上一篇    下一篇

CCL20对CBA/J小鼠CCR6+调节性T细胞及妊娠结局的影响

张晓欣，赵爱民   

  1. 上海交通大学 医学院附属仁济医院妇产科， 上海 200127
  • 出版日期:2016-02-28 发布日期:2016-03-29
  • 通讯作者: 赵爱民, 电子信箱: zamzkh0526@126.com。
  • 作者简介:张晓欣（1983—）， 女， 主治医师， 博士; 电子信箱: mxcz5293@163.com。
  • 基金资助:

    国家自然科学基金(81270715)

Effects of CCL20 on CCR6+ regulatory T cells and pregnancy outcome of CBA/J mice

ZHANG Xiao-xin, ZHAO Ai-min   

  1. Department of Obstetrics and Gynecology, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200127, China
  • Online:2016-02-28 Published:2016-03-29
  • Supported by:

    National Natural Science Foundation of China， 81270715

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摘要：

目的 研究CCL20对CBA/J小鼠CCR6+调节性T细胞(Treg细胞)及妊娠结局的影响。方法 将CBA/J（♀）×DBA/2（♂）作为自然流产小鼠模型（自然流产组），CBA/J（♀）×Balb/c（♂）作为正常妊娠小鼠模型（正常妊娠组）。于CBA/J小鼠不同的妊娠时期，观察妊娠结局，采用RT-PCR、Western blotting、ELISA、流式细胞技术检测CCL20及CCR6+Treg细胞的动态变化。给予正常妊娠小鼠CCL20中和抗体，自然流产小鼠重组CCL20，观察妊娠结局的变化。体外观察CCL20及其抗体对Treg细胞的趋化、增殖、细胞因子水平的变化。结果 妊娠14.5 d（gd 14.5），自然流产组和正常妊娠组的胚胎吸收率分别为11.25%和2.54%，两者之间差异有统计学意义（P=0.001 8）。不同妊娠时期，自然流产组小鼠外周血及子宫中CCL20及CCR6的表达均低于正常妊娠组。在妊娠早期尤其是gd 2.5时，正常妊娠组小鼠CCL20和CCR6表达水平及Treg细胞比例均明显上升，而在自然流产组小鼠中并未出现明显的上升。在gd 0.5给予正常妊娠小鼠CCL20中和抗体后，小鼠的胚胎吸收率显著增加，而CCR6+Treg的比例明显下降。给予自然流产小鼠重组CCL20后，小鼠的胚胎吸收率下降，而CCR6+Treg的比例上升。CCL20能够促进Treg细胞的趋化功能、增殖功能以及其抑制性细胞因子的分泌功能，从而导致T细胞之间的平衡向Th2及Treg细胞倾斜，有利于妊娠的维系。 结论 CBA/J（♀）×DBA/2（♂）小鼠的自然流产可能与CCL20及CCR6+ Treg的下调有关，尤其在孕早期。早期干预CCL20的蛋白水平能够改变CCR6+Treg细胞的数量及功能，影响妊娠结局。

关键词: 复发性流产, CCL20, CCR6, 调节性T细胞

Abstract:

Objective To explore the effects of CCL20 on CCR6+ regulatory T cells and pregnancy outcome of CBA/J mice. Methods The spontaneous abortion mice model CBA/J(♀) × DBA/2(♂) (spontaneous abortion group) and normal pregnant mice model CBA/J(♀) × Balb/c (♂)(normal pregnancy group) were established. The pregnancy outcomes of 2 groups were observed. Real-time PCR, Western blotting, ELISA, and flow cytometry were used to detect the dynamic changes of CCL20 and CCR6+Treg cells at different pregnant stages of CBA/J mice. Normal pregnant mice and spontaneous abortion mice were injected with CCL20 neutralizing antibody and recombined CCL20, respectively, and changes of the pregnancy outcome were observed. The effects of CCL20 and its antibody on changes of chemotaxis, proliferation, and cytokine level of Treg cells were observed in vitro. Results On gestational day 14.5 (gd 14.5), the rates of embryo resorption of spontaneous abortion group and normal pregnancy group were 11.25% and 2.54%, respectively. The difference was statistically significant (P=0.001 8). At the different pregnant stages, the expression levels of CCL20 and CCR6 in peripheral blood and uterus of rats of spontaneous abortion group were lower than those of the normal pregnancy group. At the early stage of pregnancy, especially on gd 2.5, the expression of CCL20-CCR6 and the proportion of Treg cells of normal pregnancy group significantly elevated, while those of spontaneous abortion group didnt. After normal pregnant mice were injected with CCL20 neutralizing antibody on gd 0.5, the embryo resorption rate significantly increased and the proportion of CCR6+ Treg cells significantly decreased. After spontaneous abortion mice were injected with recombined CCL20, the embryo resorption rate decreased and the proportion of CCR6+ Treg cells increased. CCL20 could increase the chemotactic function and proliferative function of Treg cells and the secretion of inhibiting cytokines, which resulted in the balance among T cells leaning towards Th2 and Treg cells and facilitated the maintenance of the pregnancy. Conclusion The spontaneous abortion of CBA/J(♀) × DBA/2(♂) mice may be relevant to the down-regulation of CCL20 and CCR6+Treg cells, especially at the early stage of pregnancy. The early intervention of the protein level of CCL20 can change the quantity and function of CCR6+Treg cells, which can influence the pregnancy outcome.

Key words: recurrent spontaneous abortion, CCL20, CCR6, regulatory T cell