针刺对胚胎着床障碍大鼠CD4^+ CD25^+ Foxp3^+调节性T细胞的影响

Effect of acupuncture on CD4^+ CD25^+ Foxp3^+ regulatory T-cells in rats of embryo implantation failure

目的:观察针刺对胚胎着床障碍大鼠CD4+CD25+Foxp3+Treg细胞的影响。方法:144只孕鼠随机分为对照组(N)、米非司酮组(M)、米非司酮+针刺组(A)和米非司酮+黄体酮组(W),每组随机再分为6 d组、8 d组和10 d组。其中M组、A组和W组妊娠1 d给予米非司酮-麻油溶液造模,N组则给予等量麻油溶液。同时,每天下午A组固定并行针刺三阴交和后三里,N组和M组仅固定,W组肌肉注射黄体酮,直到处死当天结束。统计孕鼠胚胎着床数,用流式细胞术检测外周血CD4+CD25+Foxp3+Treg细胞和子宫内膜的CD4+Foxp3+Treg细胞的比例,Western blotting和real-time PCR测定着床点的子宫内膜Foxp3的表达。结果:与N组相比,M组的着床胚胎数、外周血CD4+CD25+Foxp3+Treg细胞和子宫内膜CD4+Foxp3+Treg细胞的比例、着床点子宫内膜Foxp3蛋白和mRNA的表达均明显下降(P<0.05);与M组相比,A组和W组的着床胚胎数、外周血CD4+CD25+Foxp3+Treg细胞和子宫内膜CD4+Foxp3+Treg细胞的比例以及着床点子宫内膜Foxp3蛋白和mRNA的表达均有不同程度地升高。结论:针刺改善胚胎着床障碍大鼠的胚胎着床可能与CD4+CD25+Foxp3+Treg细胞的调节密切相关。

AIM： To observe the effect of acupuncture CD4＋CD25＋Foxp3＋ （forkhead box ≥3）＋ regulatory T-cells （Treg cells） in rats with embryo implantation failure. METHODS： One hundred and forty-four pregnant rats were randomly divided into control group （ N ）, mifepristone treatment group （ M ）, mifepristone ＋ acupuncture treatment group （A） and mifepristone ＋ progestin treatment group （W）. The rats in groups M, A and W were treated with mifepristone- sesame oil solution on day 1, while the rats in group N were injected with the same amount of sesame oil. The Housanli （ST36） and Sanyinjiao（SP6） points were selected for acupuncture. From day 1 to the time of death, the rats in group A were fasten up and then the acupuncture was performed. Accordingly, the rats in group N and group M were only fixed, and the rats in group W were given progestin. Implanted embryos in each group were counted. The proportions of CD4 ＋ CD25＋ Foxp3 ＋ Treg cells in peripheral blood and CD4＋ Foxp3 ＋ Treg cells in the endometrium were detected by flow cytometry. The mRNA and protein levels of Foxp3 were determined by real-time PCR and Western blotting, respectively. RESULTS ： Compared with group N, the number of implanted embryos, the percentages of CD4 ＋ CD25＋ Foxp3 ＋Treg cells in peripheral blood and CD4＋ Foxp3 ＋ Treg cells in the endometrium, and the expression of Foxp3 protein and mRNA in the endometrium were significantly decreased in group M （P 〈 0. 05 ）. Compared with group M, the above indexes in group A and group W were significantly increased. CONCLUSION： The effect of acupuncture in rats with embryo implantation failure may be closely correlated with the modulation of CD4 ＋ CD25＋ Foxp3 ＋ Treg cells. [