Gestrinone

Endometriosis (EMs) is a prevalent and challenging gynecological disease and current therapeutic outcomes are suboptimal. Traditional Chinese medicine (TCM) has numerous advantages in treating EMs. Bushen Wenyang Huayu Formula (BWHF), a TCM prescription, has exhibited significant clinical efficacy in treating endometriosis (EMs). However, its exact molecular mechanism remains unclear.

MicroRNA (miRNA) and oxidative stress (OS) are crucial for the occurrence and development of EMs. This study aims to explore the epigenetic mechanism underlying the efficacy of BWHF in affecting OS by altering miRNAs in EMs.

Fifty female SD rats were randomly divided into sham, model, low-dose BWHF (BWHF-L), high-dose BWHF (BWHF-H), and gestrinone groups, each consisting of 10 rats. The EMs rat model was established by autologous transplantation. Sham and model group were given with distilled water, and treatment group was given with BWHF or gestrinone. The therapeutic effects of BWHF on EMs were evaluated using ectopic lesion volume and morphology, inflammatory cytokine levels, and ROS levels. The expression of Let-7b was detected by CISH. Human endometrial stromal cells (hESCs) were treated with H₂O₂ to establish an OS cell model, followed by intervention with BWHF. hESCs migration and invasion were detected using wound healing and invasion assays. The expression of Bach1, Nrf2, and HO-1 in vivo and vitro were detected by immunohistochemistry staining, immunofluorescence staining, Western blot analysis, and qRT-PCR. The direct interaction of let-7b with the 3'-UTR of Bach1 mRNA was assessed using a dual-luciferase reporter assay.

BWHF treatment inhibited the formation of ectopic lesions in EMs rats. Mechanistically, BWHF up-regulated let-7b, leading to decreased Bach1 expression and the subsequent relief of Bach1-mediated repression on the HO-1 promoter. Furthermore, BWHF elevated Nrf2 levels and facilitated its nuclear translocation. The translocated Nrf2 then bound to the HO-1 promoter to activate transcription, thereby counteracting oxidative stress and mitigating EMs.

BWHF alleviates EMs by upregulating let-7b, which suppresses its target Bach1, thereby relieving Bach1-mediated repression of the HO-1 promoter and allowing for enhanced activation by Nrf2. This study reveals an epigenetic mechanism underlying the therapeutic effect of BWHF against EMs, providing valuable insights for advancing its clinical application.

To investigate the mechanism of Angelica sinensis polysac-charide (ASP) ameliorating endometriosis (EMT) via the Kelch-like epichlorohydrin-associated protein 1 (Keap1)/nuclear factor-erythroid 2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) pathway.

SD rats were randomly divided into six groups (n=12 per group): normal control, model control, low-dose ASP, high-dose ASP, gestrinone, and high-dose ASP+recombinant Keap1 (rKeap1). EMT models were established in all groups except the normal control group. After successful modeling, the drugs were administered once daily for 4 weeks. The mass and volume of ectopic endometrial tissue were measured. Histopathological changes were evaluated using hematoxylin and eosin staining. Levels of reactive oxygen species, malondialdehyde, and superoxide dismutase (SOD) were measured using commercial kits. The number of autophagosomes was examined under transmission electron microscopy. The relative fluorescence intensity of microtubule-associated protein light chain 3 (LC3) in endometrial tissue was assessed by immunofluorescence staining. Western blotting was used to determine the protein expression levels of LC3-Ⅱ, LC3-Ⅰ, p62, Keap1, Nrf2 in the nucleus, and HO-1.

Compared with the normal control group, the model control group showed increased glandular proliferation, pronounced vacuolization of glandular cells, and substantial inflammatory cell infiltration in ectopic endometrial tissue. Levels of reactive oxygen species, malondialdehyde, p62, and Keap1 were significantly increased, while SOD levels, autophagosome numbers, and LC3 fluorescence intensity were significantly decreased (all P<0.05). Protein expression levels of LC3-Ⅱ/LC3-Ⅰ, Nrf2 in the nucleus, and HO-1 were also significantly reduced (all P<0.05). Compared with the model control group, the low- and high-dose ASP groups and the gestrinone group exhibited alleviated pathological damages, reduced mass and volume of ectopic endometrial tissues, decreased levels of reactive oxygen species, malondialdehyde, p62, and Keap1, and increased SOD levels, autophagosome numbers, LC3 fluorescence intensity, and protein expression levels of LC3-Ⅱ/LC3-Ⅰ, Nrf2 in the nucleus, and HO-1 (all P<0.05). However, co-administration of rKeap1 reversed the inhibitory effects of high-dose ASP on oxidative stress and its restorative effects on autophagic homeostasis in EMT rats.

Angelica sinensis polysaccharide may ameliorate EMT in rats by inhibiting Keap1-mediated activation of the Nrf2/HO-1 pathway, thereby suppressing oxidative stress and restoring autophagic homeostasis.