Bushen Huoxue recipe restores trophoblast proliferation through the PI3K/AKT pathway in recurrent spontaneous abortion.

BACKGROUND

Dysregulation of extravillous trophoblast (EVT) proliferation and invasion has been implicated in the pathogenesis of recurrent spontaneous abortion (RSA). Bushen Huoxue Recipe (BSHXR), a Traditional Chinese Medicine formula for RSA, has notable clinical efficacy; however, the molecular mechanisms by which it ameliorates recurrent spontaneous abortion remain to be elucidated.

OBJECTIVE

This study aimed to identify the key active ingredients in BSHXR and observe the mechanism by which BSHXR alleviates RSA.

METHODS

BSHXR compounds were identified by UPLC-Q-TOF-MS/MS. Transcriptome sequencing and network pharmacology were performed to identify potential key genes and pathways involved in the therapeutic effects of BSHXR. A miscarriage model was established in mice, and the embryo loss rate, ELISA, and HE staining were used to explore the effect of BSHXR. In combination with bioinformatics analyses, we used immunoblotting and immunofluorescence to elucidate the regulatory mechanisms of BSHXR in vivo.

RESULTS

Transcriptome analysis suggests that the pathogenesis of RSA may be associated with cell proliferation and the PI3K/AKT pathway. A total of 155 chemical constituents of BSHXR were identified, along with 267 therapeutic targets, for treating RSA, and the main chemical components of BSHXR stably interact with key genes of the PI3K/AKT pathway. In the BSHXR group, the rate of embryonic loss was significantly reduced (p < 0.01), and the levels of serum progesterone (P) and chorionic gonadotropin (CG) increased (p < 0.05), with placental tissue morphology tending toward normalcy. Furthermore, the expression of Ki67, PCNA, and proteins related to the PI3K/AKT (p < 0.0001) pathway increased in the BSHXR group.

CONCLUSION

The PI3K/AKT pathway is a potential pathway for RSA intervention. The therapeutic effect of BSHXR is related to the proliferative effect mediated by the PI3K/AKT pathway.