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OA

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 β2 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,total 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.