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补肾活血方改善抗磷脂抗体相关复发性流产小鼠妊娠结局的机制
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| 篇名： | 补肾活血方改善抗磷脂抗体相关复发性流产小鼠妊娠结局的机制 |
| TITLE： | Mechanism of Bushen huoxue formula in improving pregnancy outcomes in mice with antiphospholipid antibody-related recurrent spontaneous abortion |
| 摘要： | 目的 探讨补肾活血方改善抗磷脂抗体相关复发性流产（aPL-RSA）小鼠妊娠结局的作用机制。方法将SPF级BALB/c雌性小鼠随机分为正常对照组、佐剂对照组、模型组和补肾活血方组。以β2糖蛋白Ⅰ联合弗氏佐剂免疫法构建小鼠aPL-RSA模型。补肾活血方组自妊娠第0天起灌胃给药1.6536g/（kg·d），其余3组同期灌胃等体积生理盐水，每日2次，连续给药15d。观察孕鼠子宫外观，记录单个胚胎质量并计算胚胎吸收率；检测富血小板血浆（PRP）中CD41+CD62p+活化血小板比例；检测血清中血栓素B2（TXB2）、血小板因子4（PF4）水平；观察胎盘组织形态变化；检测胎盘组织细胞凋亡率；检测胎盘组织中丙二醛（MDA）含量、总超氧化物歧化酶（T-SOD）活性以及白细胞介素1β（IL-1β）、IL-18水平，基质金属蛋白酶3（MMP-3）、MMP-9、增殖细胞核抗原-67（简称Ki67）、超氧化物歧化酶2（SOD2）、Nod样受体蛋白3（NLRP3）、含有CARD结构域的凋亡相关斑点样蛋白（ASC）、胱天蛋白酶1（caspase-1）蛋白表达和SOD2、NLRP3、ASC、caspase-1mRNA表达。结果与正常对照组和佐剂对照组比较，模型组孕鼠的单个胚胎质量、胎盘组织中T-SOD活性，以及SOD2mRNA和蛋白的相对表达量均显著降低（P＜0.05）；胚胎吸收率，PRP中CD41+CD62p+活化血小板比例，血清中TXB2、PF4水平，胎盘组织细胞凋亡率和胎盘组织中MDA含量，MMP-3、MMP-9、Ki67蛋白的相对表达量，NLRP3、ASC、caspase-1蛋白和mRNA的相对表达量，IL-1β、IL-18水平均显著升高（P＜0.05）；模型组孕鼠子宫形态不规则，部分子宫角存在局限性萎缩；胎盘组织蜕膜细胞碎裂，迷路区滋养层细胞广泛空泡化、坏死，伴血管减少。经补肾活血方干预后，孕鼠上述指标水平均显著逆转（P＜0.05），孕鼠子宫形态、病理损伤均明显改善。结论补肾活血方可有效改善aPL-RSA模型孕鼠的妊娠结局，其作用机制可能与抑制血小板活化、减轻氧化应激损伤、增强滋养层细胞功能，以及抑制NLRP3炎症小体通路介导的炎症反应密切相关。 |
| ABSTRACT： | OBJECTIVE To investigate the mechanism of Bushen huoxue formula in improving pregnancy outcomes in mice with antiphospholipid antibody-related recurrent spontaneous abortion （aPL-RSA）. METHODS SPF female BALB/c mice were randomly divided into normal control group， adjuvant control group， model group， and Bushen huoxue formula group. The aPL-RSA mouse model was established by immunization with β 2 glycoproteinⅠcombined with Freund’s adjuvant. From gestational day 0， the Bushen huoxue formula group was administered 1.653 6 g/（kg·d） of the prescription by gavage， while the other three groups received an equal volume of normal saline， twice daily for 15 consecutive days. The uterine appearance of pregnant mice was observed； individual embryo weight was recorded， and the embryo resorption rate was calculated. The proportion of activated CD41 + CD62p + platelets in platelet-rich plasma （PRP） was detected. Serum levels of thromboxane B2 （TXB2） and platelet factor 4 （PF4） were measured. Morphological changes of placental tissue were observed. The cell apoptosis rate of placental tissue was detected. The levels of malondialdehyde （MDA） content， tot al superoxide dismutase （T-SOD） activity， the interleukin-1β （IL-1β） and IL-18， the protein expressions of matrix metalloproteinase-3 （MMP-3）， MMP-9， nuclear proliferation antigen-67 （Ki67）， superoxide dismutase 2 （SOD2）， Nod-like receptor protein 3 （NLRP3）， apoptosis-associated speck-like protein containing a CARD （ASC）， and caspase-1， as well as the mRNA expressions of superoxide dismutase 2（SOD2）， NLRP3， ASC and caspase-1 of placental tissue were detected. RESULTS Compared with the normal control group and adjuvant control group， the model group showed significant decreases in individual embryo weight， T-SOD activity of placental tissue， and relative expression levels of SOD2 mRNA and protein （ P ＜0.05）； while significant increases were observed in embryo resorption rate， proportion of activated CD41 + CD62p + platelets in PRP， serum levels of TXB2 and PF4， placental cell apoptosis rate， MDA content in placental tissue， relative protein expressions of MMP-3， MMP-9 and Ki67， relative protein and mRNA expression levels of NLRP3， ASC and caspase-1 as well as IL-1β and IL-18 levels （ P ＜0.05）. The model group also exhibited irregular uterine morphology with localized atrophy in some uterine horns； placental tissue showed fragmentation of decidual cells， extensive vacuolization and necrosis of trophoblast cells in the labyrinthine zone， accompanied by vascular reduction. After intervention with Bushen huoxue formula， the above indicators were significantly reversed （ P ＜0.05）， and uterine morphology and pathological damage were markedly improved. CONCLUSIONS Bushen huoxue formula can effectively improve pregnancy outcomes in aPL-RSA model mice， and its mechanism may be closely related to inhibiting platelet activation， reducing oxidative stress injury， enhancing trophoblast cell function， and suppressing the inflammatory response mediated by the NLRP3 inflammasome pathway. |
| 期刊： | 2026年第37卷第11期 |
| 作者： | 韩永梅；崔天薇；谢俊丹；何鑫雨；龚宇婷；王满满 |
| AUTHORS： | HAN Yongmei，CUI Tianwei，XIE Jundan，HE Xinyu，GONG Yuting，WANG Manman |
| 关键字： | 补肾活血方; 复发性流产; 抗磷脂抗体; 血小板活化; NLRP3炎症小体; 妊娠结局 |
| KEYWORDS： | Bushen huoxue formula; recurrent spontaneous abortion; antiphospholipid antibody; platelet activation; NLRP3 inflammasome; pregnancy outcome |
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