Effect of Modified Shaofu Zhuyutang on Ferroptosis in Ectopic Endometrial Tissues of Rats with Endometriosis Based on MDM4/p53/GPX4 Signaling Pathway
10.13422/j.cnki.syfjx.20242245
- VernacularTitle:基于MDM4/p53/GPX4信号通路探讨加味少腹逐瘀汤对子宫内膜异位症大鼠异位子宫内膜组织铁死亡的影响
- Author:
Zuoliang ZHANG
1
;
Xiangyu LIN
1
;
Wanrun WANG
1
;
Jiaxing WANG
1
;
Yaling YANG
2
;
Quansheng WU
1
Author Information
1. Clinical College of Chinese Medicine， Gansu University of Chinese Medicine， Lanzhou 730000，China
2. Affiliated Hospital of Gansu University of Chinese Medicine，Lanzhou 730030，China
- Publication Type:Journal Article
- Keywords:
endometriosis;
ferroptosis;
modified Shaofu Zhuyutang;
murine double minute 4/tumor suppressor p53/glutathione peroxidase 4 （MDM4/p53/GPX4） signaling pathway
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2025;31(4):39-47
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo investigate the mechanism of modified Shaofu Zhuyutang in inducing ferroptosis in ectopic endometrial tissues of rats with endometriosis through the murine double minute 4 （MDM4）/tumor suppressor p53/glutathione peroxidase 4 （GPX4） signaling pathway. MethodsSeventy SPF-grade female SD rats were randomly divided into a blank group （n = 10）， a sham-operated group （n = 10）， and a modeling group （n = 50）. The sham-operated group underwent laparotomy， while the modeling group was subjected to the autotransplantation method to establish an endometriosis model. After successful modeling， the animals were randomly assigned to the model group， progesterone group （0.25 mg·kg-1）， and modified Shaofu Zhuyutang high-， medium-， and low-dose groups （30， 15， and 7.5 g·kg-1， respectively）， with 10 rats per group. After four weeks of drug administration， the rats were euthanized for sample collection. The weight and volume of ectopic endometrial tissues were recorded for each group. Transmission electron microscopy （TEM） was employed to observe ultrastructural changes in endometrial tissues， while Prussian blue staining was used to assess iron ion deposition. Serum levels of interleukin-6 （IL-6）， interleukin-1β （IL-1β）， and tumor necrosis factor-α （TNF-α） were measured by enzyme-linked immunosorbent assay （ELISA）. The relative levels of Fe2+， malondialdehyde （MDA）， and glutathione （GSH） in endometrial tissues were determined by colorimetric assay. Immunofluorescence （IF） was used to detect the relative fluorescence intensities of GSH and GPX4 in endometrial tissues. The relative expression levels of phosphatidylinositol 3-kinase （PI3K）， phosphorylated PI3K （p-PI3K）， protein kinase B （Akt）， phosphorylated Akt （p-Akt）， MDM4， p53， and GPX4 proteins were detected by Western blot. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to assess the mRNA expression of PI3K， Akt， MDM4， p53， and GPX4. ResultsCompared with the blank and sham-operated groups， the model group exhibited reduced ferroptotic damage in ultrastructural observations， decreased ferroptotic aggregates and positive iron ion expression area on Prussian blue staining， elevated serum IL-6， IL-1β， and TNF-α levels， reduced Fe2+ and MDA content， increased GSH content in endometrial tissues， and enhanced GSH and GPX4 fluorescence intensities （P<0.01）. The protein and mRNA expression levels of PI3K， p-PI3K， Akt， p-Akt， MDM4， and GPX4 were elevated， while those of p53 were decreased （P<0.01）. Compared with the model group， in the progesterone group and the modified Shaofu Zhuyutang high- and medium-dose groups， ferroptotic damage in ultrastructural observations was exacerbated to varying degrees by TEM， and ferroptotic aggregates and positive iron ion expression areas were increased on Prussian blue staining. Serum IL-6， IL-1β， and TNF-α levels decreased， Fe2+ and MDA content increased， and GSH content decreased in endometrial tissues. GSH and GPX4 fluorescence intensities weakened， while the protein and mRNA expression levels of PI3K， p-PI3K， Akt， p-Akt， MDM4， and GPX4 decreased， and those of p53 increased （P<0.05， P<0.01）. Conclusionmodified Shaofu Zhuyutang may exert therapeutic effects in endometriosis by inducing ferroptosis in ectopic endometrial tissues， alleviating inflammatory responses， and modulating key proteins in the MDM4/p53/GPX4 signaling pathway.
