抗心磷脂抗体导致妊娠丢失的免疫病理机制及安子合剂的干预作用

Study on the Immunopathogenesis of Anticardiolipin Antibody Induced Fetal Loss and the Interference of Anzi Heji

目的:探讨抗心磷脂抗体(ACA)导致妊娠丢失与CD4+CD25+FOXP3+调节性T(CD4+CD25+FOXP3+Treg)细胞的关系及安子合剂的免疫调节机制。方法:将BALB/c小鼠随机分组:空白组、模型组、安子合剂组(低、中、高剂量组)、阿司匹林组。于妊娠第1天,空白组和模型组以等量蒸馏水灌胃,安子合剂组(低、中、高剂量组分别以37.7,75.4,150.8 g.kg-1.d-1)药液ig,阿司匹林组以0.019 5 g.kg-1.d-1ig,连续给药14 d。于妊娠第8,12天通过给孕鼠腹腔注射抗心磷脂抗体-IgG(ACA-IgG),建立ACA阳性妊娠丢失动物模型。用流式细胞仪测定各组孕鼠外周血CD4+CD25+FOXP3+Treg细胞的比例;用ELISA法测定各组孕鼠外周血ACA的滴度;并观察各组孕鼠胚胎吸收及胎鼠发育情况。结果:模型组CD4+CD25+FOXP3+Treg细胞的比例较空白组显著下降(2.67±0.51)%,(9.849±1.774)%,P<0.01;与模型组相比,安子合剂低、中剂量组均能显著升高CD4+CD25+FOXP3+Treg细胞的比例(9.47±1.26),(7.61±1.07)%,P<0.01,显著降低ACA的滴度(P<0.01),安子合剂低剂量组优于中剂量组。安子合剂低剂量组能显著降低胚胎吸收率,显著增加胎鼠和胎盘质量(P<0.05)。结论:ACA导致妊娠丢失的病理机制与免疫调节细胞-CD4+CD25+FOXP3+Treg细胞比例减少有关。安子合剂干预ACA导致妊娠丢失的免疫调节机制是通过增加CD4+CD25+FOXP3+Treg细胞的比例来实现的,其作用的强弱与药物的剂量相关。

Objective： To explore the immune pathological mechanism of anticardiolipin antibody （ACA） induced fetal loss and the immune regulation mechanism of Anzi Heji. Method： BALB/c mice were randomly divided into 6 groups： control group, model group, low dose group, middle dose group and high dose group of Anzi Heji, aspirin group. At the first day of pregnancy, distilled water was orally given in control group and model group, while the mice in Anziheji groups were given Anziheji groups at dose of 37.7, 75.4, 150. 8 g·kg^-1·d^-1 respectively, the mice in aspirin group were given aspirin with 0. 019 5 g·kg^-1·d^-1. The administration was given for 14 days, once a day. ACA-IgG injected to pregnant mice by intraperitoneal injection to establish animal model of ACA induced fetal loss on the day 8 and day 12 of pregnancy. The peripheral blood CD4 ~ CD25 ~ FOXP3 ~ Treg cells ratio was detected by flow cytometry; ACA level of mice pregnancy was detected by ELISA; the embryo resorption and fetal development situation were observed. Result： In model group, the peripheral blood CD4＋ CD25 ＋ FOXP3 ＋ Treg cells ratio was significantly lower than the control group （2.67 ± 0.51 ）% vs （9.85 ± 1.77） % （P 〈 0. 01）; compared with model group, the low and middle dose group of Anzi Heji could increase CD4＋CD25＋FOXP3＋Tregcells ratio significantly （9.47±1.26）%, （7.61±1.07）% vs （2.67±0.51）%, P 〈 0.01, and significantly reduce the titer of ACA （P 〈 0.01 ） , the low dose group of Anzi Heji was better than the middle dose group of Anzi Heji （P 〈0.01）. The low dose group of Anzi Heji could significantly reduce the resorption rate, and significantly increased fetal weight and placental weight （P 〈 0.05 ）. Conclusion： Pathological mechanism of ACA induced fetal loss is related to decreasing immune cells-CD4 ＋ CD25 ＋ FOXP3 ＋ Treg cells percentage. The immune regulatory mechanism of Anzi Heji on ACA induced fetal loss can be achieved by increasing the CD4 ＋ CD25 ＋ FOXP3 ＋ Treg cells ratio.