【摘要】 目的 本研究旨在通过构建动物实验模型探讨何氏补肾益气活血方对抗心磷脂抗体（ACA）阳性血栓前状态复发性流产（RSA）的干预效应，并基于酪氨酸蛋白激酶/信号转导子和转录激活子3(JAK/STAT3)信号传导途径阐明其可能的作用机制。方法 BALB/c雌性小鼠50只，随机分为空白对照组（蒸馏水灌胃）、模型对照组（蒸馏水灌胃）、阿司匹林组（9.75 g/kg灌胃）及何氏补肾益气活血方低、高剂量（中药等效剂量37.7 g/kg、75.4 g/kg灌胃）组，每组10只。采用人源β2-糖蛋白Ⅰ（β2-GPⅠ）构建ACA阳性RSA小鼠模型。从妊娠第1天开始灌胃给药，干预14 d。处死小鼠，留取样本，测定胚胎吸收率。通过酶联免疫吸附法（ELISA）定量分析外周血ACA水平；应用免疫组化检测母胎界面（胎盘及蜕膜）非受体酪氨酸蛋白激酶（JAK）、糖蛋白130(GP130)、磷酸化信号转导子与转录激活子3(p-STAT3)及增殖细胞核抗原（PCNA）表达；采用蛋白质印迹法（Western blot）定量分析母胎界面STAT3磷酸化水平。结果 与模型组比较，何氏补肾益气活血方低、高剂量组及阿司匹林组胚胎丢失率[（24.49±11.17）%、（16.18±11.92）%、（15.94±8.01）%比（42.53±15.12）%,P<0.05]、血清ACA滴度[（93.16±6.75）pg/mL、（92.31±5.22）pg/mL、（81.60±5.18）pg/mL比（419.51±94.00）pg/mL,P<0.05]显著降低。与模型组比较，何氏补肾益气活血方低、高剂量组及阿司匹林组母胎界面胎盘JAK[(1.44±0.53)、（1.43±0.50）、（1.51±0.35）比（0.21±0.31）,P<0.05]、胎盘p-STAT3[(1.83±0.41)、（2.00±0.63）、（2.00±0.71）比（0.20±0.45）,P<0.05]、蜕膜p-STAT3[(1.50±0.54)、（2.13±0.42）、（1.80±0.65）比（0.40±0.38）,P<0.05]、胎盘PCNA[(2.50±0.55)、（2.67±0.52）、（2.80±0.84）比（1.39±0.99）,P<0.05]及蜕膜PCNA[(2.88±0.36)、（3.00±0.58）、（2.91±0.68）比（1.40±0.98）,P<0.05]表达显著上调。何氏补肾益气活血方低、高剂量组及阿司匹林组蜕膜及胎盘p-STAT3/蛋白表达的灰度积比值[（1.05±0.08）、（1.05±0.07）、（1.06±0.09）比（0.89±0.09）,P<0.05,(1.07±0.09)、（1.09±0.07）、（1.09±0.09）比（0.79±0.06）,P<0.05]显著高于模型组。结论 何氏补肾益气活血方通过降低ACA滴度，激活JAKSTAT3信号通路并促进细胞增殖，改善ACA阳性血栓前状态RSA小鼠的胚胎着床环境以及妊娠结局。更多还原

【Abstract】 Objective To investigate the therapeutic effects of He’s Bushen Yiqi Huoxue Decoction on recurrent spontaneous abortion(RSA) in an antinticardiolipin antibody(ACA) prothrombotic state by establishing an animal experimental model, and to elucidate its potential mechanism based on Janus kinase(JAK)/signal transducer and activator of transcription 3(STAT3) signaling pathway. Methods Fifty female BALB/c mice were randomly divided into five groups(n=10 per group): blank control group(gavage with distilled water), model control group(gavage with distilled water), aspirin group(gavage with 9.75 g/kg aspirin), and low-(gavage with herbal equivalent doses of 37.7 g/kg) and high-dose(gavage with herbal equivalent doses of 75.4 g/kg) He’s Bushen Yiqi Huoxue Decoction groups. The ACA-positive RSA mouse model was established using human β2-glycoprotein I. Pregnant mice received daily gavage interventions for 14 consecutive days from gestational day 1. Following euthanasia, the embryo resorption rate was determined, and the blood and tissue samples were collected. Peripheral blood ACA levels were quantitatively analyzed using enzyme-linked immunosorbent assay. The expression levels of JAK, glycoprotein130(GP130), phosphorylated STAT3(p-STAT3), and proliferating cell nuclear antigen(PCNA) at the maternal-fetal interface(decidua and placenta) were analyzed using immunohistochemical staining. p-STAT3 levels in the same tissue compartments were evaluated using western blotting. Results Compared with the model control group, the aspirin group, low-and high-dose He’s Bushen Yiqi Huoxue Decoction groups demonstrated significantly lower embryo loss rates[(24.49±11.17)%,(16.18±11.92)%, and(15.94±8.01)% vs.(42.53±15.12)%, respectively; P<0.05]and significantly lower serum ACA titers [(93.16±6.75)pg/mL,(92.31±5.22)pg/mL, and(81.60±5.18)pg/mL vs.(419.51±94.00)pg/mL; P<0.05]. Compared with the model control group, the low-dose and high-dose He’s Bushen Yiqi Huoxue Formula groups, along with the aspirin group, demonstrated significantly increased expression levels of placental JAK[(1.44±0.53),(1.43±0.50),(1.51±0.35) vs.(0.21±0.31); P<0.05], placental p-STAT3[(1.83±0.41),(2.00±0.63),(2.00±0.71) vs.(0.20±0.45); P<0.05], decidual p-STAT3[(1.50±0.54),(2.13±0.42),(1.80±0.65) vs.(0.40±0.38); P<0.05], placental PCNA[(2.50±0.55),(2.67±0.52),(2.80±0.84) vs.(1.39±0.99); P<0.05], and decidual PCNA[(2.88±0.36),(3.00±0.58),(2.91±0.68) vs.(3.50±0.59); P<0.05] at the maternal-fetal interface. Compared with the model group, the low-dose and high-dose He’s Bushen Yiqi Huoxue Formula groups and the aspirin group exhibited significantly higher p-STAT3/STAT3 protein expression ratios in decidua[(1.05±0.08),(1.05±0.07),(1.06±0.09) vs.(1.06±0.09); P<0.05] and placenta[(1.07±0.09),(1.09±0.07),(1.09±0.09) vs.(0.79±0.06);P<0.05]. Conclusion He’s Bushen Yiqi Huoxue Decoction improves the embryonic implantation microenvironment and pregnancy outcomes in ACA-positive prothrombotic RSA mice by reducing ACA titers, activating the JAKSTAT3 signaling pathway, and promoting cellular proliferation.更多还原