Therapeutic effect of Bushen Huoxuetfang on recurrent abortion induced by cardiolipin antibody in rats
OBJECTIVE To investigate the therapeutic effect of Bushen Huoxuefang on recurrent abortion induced by cardiolipin antibody(CL)in rats and its effect on the platelet/purinergic2X7(P2X7)/Nod-like receptor protein 3(NLRP3)inflammatory vesicle pathway.METHODS Twenty-four pregnant rats were randomly divided into aCL-RSA group,aCL-RSA+LMWH group,aCL-RSA+Danshou tang group,and NH-IgG group,with 6 rats in each group.The aCL-RSA pregnant mouse model was established,and Bushen Huoxuefang was given intervene,and the body mass of the pregnant rats,embryo absorption rate and fetal body mass were observed in E16 days.The contents of platelet-activating factor receptor(PAFR)and platelet factor 4(PF4)in rat plasma,interleukin-1β(IL-1β)and IL-18 in rat placental tissue were detected by ELISA,the histopathological changes of placental tis-sue were detected by HE staining,and apoptosis in placental tissue was detected by TUNEL staining.Western Blot analysis was performed to detect the expression of cleaved caspase-1,human apoptosis-associated mottle pro-tein(ASC),NLRP3,Bcl2,Bax,cleaved caspase-3,and P2X7 proteins,and the correlation between APL-induced recurrent abortion and platelet activation,NLRP3 inflammasome pathway,and purinergic2X7(P2X7),as well as the effect of Danshou decoction on aCL-RSA pregnant mice and platelet activation/P2X7/NLRP3 pathway,were investigated.RESULTS Compared with NH-IgG group,the aCL-RSA group could significantly reduce the body mass of pregnant rats,increase embryo absorption rate of pregnant rats,reduce the body mass of fetal rats,and increase the plasma PAFR and PF4 contents of pregnant mice(P＜0.05).The aCL-RSA group had fewer maternal erythrocytes more thrombosis in the labyrinthine area of placental tissue,and a significantly reduced placental blood sinus space.There was a significant increase in the number of apoptotic cells,an increased in IL-1β and IL-18,and an increase in the expressions of cleaved caspase-1,ASC,NLRP3,Bax,cleaved caspase-3,and P2X7 in placenta tissues(P＜0.05),and a decrease in the level of Bcl2 protein(P＜0.05).CONCLUSION Bushen Huox-uefang could inhibit platelet activation,down-regulate the expression of P2X7,inhibit the activation of NLRP3,elevate the hypercoagulability and inflammation of placenta,and then reduce the occurrence of aCL-RSA.
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