当归对诱导性流产小鼠子宫局部细胞免疫的影响

Effect of Angelica sinensis on local cellular immunity in uterus of induced abortion mice

为了探讨保胎中药当归对流产小鼠的保胎效果,及当归对妊娠小鼠子宫局部细胞免疫的影响,试验将小鼠通过自然交配发生妊娠,将怀孕小鼠随机分为3组,每组10只,其中两组使用米非司酮(RU486)诱导小鼠流产,制作试验小鼠流产模型,即空白对照组(A组)小鼠于孕第1~7天,每天灌服纯化水,0.5 mL/只,在孕第7天,经颈部皮下注射丙二醇0.1 mL/只;流产模型组(B组)小鼠于孕第1~7天每天灌服纯化水,0.5 mL/只,在孕第7天,经颈部皮下注射RU486,0.1 mL(0.025 mg)/只;当归治疗组(C组)小鼠于孕第1~7天每天灌服当归水煎剂,0.5 mL(相当于0.5 g生药)/只,在孕第7天,经颈部皮下注射RU486,0.1 mL/只。通过统计流产小鼠胚胎丢失比例,评价中药的保胎效果;采用流式细胞技术检测流产与成功妊娠小鼠子宫组织中Th17细胞、Treg细胞及比例,并测定小鼠子宫组织中白细胞介素-6(IL-6)、白细胞介素-10(IL-10)及转化生长因子-β(TGF-β)的含量。结果表明:RU486造模后,C组小鼠流产率下降至40%,胚胎吸收率减少至34.92%,子宫组织中Th17细胞比例为(1.52±0.30)%,与B组比较极显著降低(P<0.01);Treg细胞比例为(10.21±1.18)%,与模型组比较极显著升高(P<0.01);Th17/Treg值为0.15,极显著低于B组(P<0.01);子宫组织中IL-6含量为(5.30±1.16)ng/mL,极显著低于B组(P<0.01);IL-10含量为(122.20±43.15)pg/mL,极显著高于B组(P<0.01);但TGF-β含量与B组差异不显著(P>0.05)。说明中药当归可以调节子宫局部免疫细胞的分化及一些与妊娠相关的细胞因子的含量,调整妊娠子宫局部免疫状态,通过对子宫局部的免疫调节促进Treg细胞分化,并通过Treg介导的免疫抑制,使免疫有利于保持妊娠过程。

In order to explore the fetal-protecting effect of Chinese medicine Angelica sinensis on abortion mice,and the effect of Angelica sinen⁃sis on local cellular immunity in uterus of mice during pregnancy,pregnancy occurred in mice through natural mating,and mifepristone(RU486)was used to induce abortion in mice to make experimental animal abortion model.Pregnant mice were randomly divided into 3 groups of 10 mice each.Mice in the blank control group(group A)were given purified water every day,0.5 mL per mice on the 1 st to 7 th day of pregnancy,and 0.1 mL per mice was subcutaneously injected with propylene glycol through the neck on the 7 th day of pregnancy.Mice in the abortion model group(group B)were given purified water daily on the 1 st to 7 th day of pregnancy,0.5 mL per mice,and on the 7 th day of pregnancy,RU486 was subcutaneously injected through the neck,0.1 mL(0.025 mg)per mice.Mice in Angelica sinensis treatment group(group C)were given Angelica decoction,0.5 mL(equivalent to 0.5 g crude drug)per mice every day from the 1 st to 7 th day of pregnancy,and on the 7 th day of pregnancy,RU486 was injected subcutaneously through the neck,0.1 mL per mice.To count the proportion of embryos lost in aborted mice,the fetal-protecting effect of traditional Chinese medicine was evaluated.Flow cytometry was used to detect the proportion of Th17 cells and Treg cells in the uterine tissue of aborted and successfully pregnant mice,and the contents of interleukin-6(IL-6),inter⁃leukin-10(IL-10)and transforming growth factor-β(TGF-β)in mouse uterine tissue were determined.The results showed that after RU486 model was established,the abortion rate of the mice in the group C decreased to 40%,the embryo absorption rate decreased to 34.92%,and the proportion of Th17 cells in the uterine tissue was(1.52±0.30)%,which was significantly lower than group B(P<0.01).The proportion of Treg cells was(10.21±1.18)%,which was significantly higher than that of the model group(P<0.01).Th17/Treg value was 0.15,which was significantly lower than group B(P<0.01);IL-6 content in uterine tissue was(5.30±1.16)ng/mL,which was significantly lower than group B(P<0.01);the content of IL-10 reached(122.20±43.15)pg/mL,which was significantly higher than that in group B(P<0.01),but the content of TGF-βwas not significantly different from that in group B(P>0.05).The results suggested that Chinese medicine Angelica sinensis could regulate the differentiation of local immune cells in the uterus and the contents of some pregnancy-related cytokines,and adjust the local immune status of the pregnant uterus.By local immunomodulation of the uterus,the immune response was biased towards Treg cells,and through Treg-mediated immunosuppression,immunity was favored to maintain the pregnancy process.