GuBen AnTai decoction ameliorates recurrent spontaneous abortion by suppressing excessive autophagy via EGFR/PI3K...

The Chinese herbal formula Guben Antai Decoction (GBAT) demonstrates efficacy in reducing pregnancy loss associated with recurrent spontaneous abortion (RSA). However, the molecular mechanisms underlying its impact on the physiological functions of the placental trophoblast cells remain unclear.

The objectives were to assess GBAT's efficacy in both a trophoblast-cell oxidative-stress model and a pregnant mouse model of RSA and to elucidate the complex mechanisms underlying the prevention of pregnancy loss using comprehensive multi-omics analysis integrated with a network pharmacology framework.

Network pharmacology and bioinformatics were used to screen key GBAT targets associated with regulating autophagy and improving RSA. We established an RSA pregnant mouse model (CBA/J x DBA/2) comprising normal, RSA, low-dose GBAT (L-GBAT), high-dose GBAT (H-GBAT), and Dydrogesterone (DYD) groups. The maternal-fetal interface was analyzed at the autophagy level by performing HE staining, immunohistochemistry, transmission electron microscopy, and western blotting. We treated HTR-8/Svneo cells with H₂O₂ (40 μM, 3 h) to establish a trophoblast cell oxidative damage model with the following groups: control, H₂O₂, H₂O₂+GBAT, H₂O₂+GBAT+RAPA (autophagy activator), and H₂O₂+GBAT+Gefitinib (EGFR inhibitor). We performed drug-containing serum interventions in these groups and examined GBAT's modulation of autophagic activity and EGFR/PI3K phosphorylation via western blotting, MDC assay, and immunofluorescence.

Network pharmacology and bioinformatics analyses showed that the EGFR/PI3K signaling pathway was an important mediator of GBAT-induced RSA autophagy. In vivo, GBAT inhibited autophagy and downregulated EGFR/PI3K phosphorylation in the Placental tissue placental villi of the RSA mice, which improved pregnancy outcomes. In vitro, GBAT-containing serum inhibited autophagy in oxidatively stressed HTR-8/Svneo cells. This effect was partially reversed by RAPA or gefitinib treatment.

GBAT may effectively inhibit excessive autophagic activation in RSA. The EGFR/PI3K pathway plays an important role in its therapeutic efficacy. Thus, GBAT is a promising candidate for alleviating RSA-induced miscarriage.