Deciphering the Mechanism of Bushen Huoxue Decotion on Decidualization by Intervening Autophagy via AMPK/mTOR/ULK1: A Novel Discovery for URSA Treatment

Abstract

Impaired decidualization was recognized as one of the crucial pathomechanisms accounting for unexplained recurrent spontaneous abortion (URSA). Currently, the exact molecular mechanism and targeted clinical decision are still in exploration. Bushen Huoxue decoction (BSHXD) has previously been proved effective in treating URSA, but its mechanism remains to be elucidated. This study aimed to explore the regulation mechanism of BSHXD in decidualization from its intervention in autophagy so as to rationalize its potential as a novel therapeutic regime for URSA. Decidua tissues were collected from patients with URSA and healthy pregnant women who underwent legal terminations for non-medical reasons at the first trimester. Besides, cell line T-hESCs was utilized to establish induced decidualization model, and were randomly divided into ESC group, DSC group, 3-MA group, AMPK siRNA group, scrambled siRNA group and AMPK siRNA + BSHXD group. Transmission electron microscopy, Monodansylcadaverine (MDC) assay, qRT-PCR, immunohistochemistry, immunofluorescence and Western blotting were used to evaluate the level of decidualization, autophagy and activation of AMPK signaling pathway in decidua tissues and cell experiments. Experiments on decidua tissues showed that decidualization was impaired in URSA with inhibited autophagy. Besides, pAMPK T172 and pULK1 S556 were decreased, and pmTOR S2448 and pULK1 S757 were increased. Cell experiments showed that the level of autophagy increased during induced decidualization, but when autophagy was inhibited, decidualization was impaired. In addition, AMPK/mTOR/ULK1 affected decidualization by mediating autophagy, and BSHXD improved decidualization through this mechanism. In conclusion, this study clarified that the inhibition of autophagy mediated by AMPK/mTOR/ULK1 was associated with impaired decidualization, and the intervention of BSHXD on this pathological process may be a vital mechanism for its treatment of URSA. This study laid the foundation for further research and application of BSHXD.

Intercellular communication involving macrophages at the maternal-fetal interface may be a pivotal mechanism of URSA: a novel discovery from transcriptomic data

“…As for the validation of signature genes decidua tissues were fixed with 4% paraformaldehyde and embedded in paraffin for immunofluorescence analysis, which was conducted following previously described methods ( 29 ). In this study, Vimentin representative decidual stromal cell and CX3CR1 representative macrophage.…”

Modulating Morphological and Redox/Glycative Alterations in the PCOS Uterus: Effects of Carnitines in PCOS Mice

“…Western blotting was used to detect the expression levels of LC3II/LC3I and p62, the two of which can be used to evaluate the status of autophagy flux [34]. The results showed that the LC3II/LC3I ratio in the DHEA group decreased significantly, while p62 increased (Figure 5), indicating the decreased level of autophagy in the DHEA uterus.…”

“…Autophagy is known to drive essential homeostatic mechanisms in uterus physiology and has been involved in uterus hyperplasia [33]. To quantify the level of autophagy, Western blotting was used to detect the expression levels of LC3II/LC3I and p62, the two of which can be used to evaluate the status of autophagy flux [34]. The results showed that the LC3II/LC3I ratio in the DHEA group decreased significantly, while p62 increased (Figure 5), indicating the decreased level of autophagy in the DHEA uterus.…”