Huayu Jiedu Fang Protects Ovarian Function in Mouse with Endometriosis Iron Overload by Inhibiting Ferroptosis

Abstract

Endometriosis (EM) is a common chronic inflammatory disease in women. Sampson’s retrograde menstruation theory is the most widely accepted theory of EM pathogenesis. The periodic bleeding of ectopic lesions is an important pathological feature of this disease, and the occurrence and progression of EM are closely associated with the iron overload caused by ectopic lesions. However, animal models that simulate menstrual-blood reflux and hemorrhage from EM lesions are lacking. In this study, we performed intraperitoneal injection of endometrial fragments and periodic intraperitoneal blood injection to simulate the real cause and disease state of EM and successfully constructed a mouse model of EM iron overload. Our research found that the number, size, and degree of adhesion of EM lesions in the iron-overload model mouse were significantly higher than those in the model mouse. Moreover, the iron concentration in the abdominal fluid and ovary significantly increased, and the level of malondialdehyde (MDA) in the ovary increased. Conversely, GPX4, GSH, and other anti-ferroptosis-related proteins were downregulated, proving the occurrence of ferroptosis. Huayu Jiedu Fang (HYJDF) is an empirical prescription for EM treatment. This study combined animal experiments, UHPLC-QE-MS analysis, and network pharmacology to analyze whether HYJDF can inhibit ferroptosis to slow down the progression of EM and protect ovarian function. Based on the constructed iron-overload model, HYJDF can reduce the volume of EM lesions and the degree of adhesion, downregulate the total iron concentration in the peritoneal fluid and ovary, upregulate GPX4 expression and GSSG in the ovary, downregulate the level of MDA in the ovary, and promote the development of follicles. We further confirmed that HYJDF can inhibit the progression of EM disease and improve the ovarian function of the model mouse by inhibiting ferroptosis. Finally, through UHPLC-QE-MS and network pharmacology analysis, the natural compounds in HYJDF were identified and verified and the regulatory effect of HYJDF on the EM ferroptosis pathway through the IL-6/hepcidin pathway was preliminarily elucidated.

Mechanism of ferroptosis in a rat model of premature ovarian insufficiency induced by cisplatin

“…When SLC7A11, the major functional subunit of system Xc − , is deficient or has reduced activity, GSH synthesis is reduced, causing reduced expression and activity of GPX4, resulting in the inability of cells to effectively scavenge lipid peroxides, and finally oxidative damage to the cell membrane, leading to cell ferroptosis 42 . Previous studies have reported a correlation between GSH, GPX4 and ovarian dysfunction; however, few studies have focused on the correlation between SLC7A11 and POI [43][44][45][46] . Therefore, we mainly selected SLC7A11 to further validate the association of POI and ferroptosis, but we still validated the expression of GPX4 in animal experiments.…”

Role of autophagy and ferroptosis in the development of endometriotic cysts (Review)

“…Excess iron decreases the expression of solute carrier family 7 member 11 (SLC7A11; a light-chain subunit of xCT) and GPX4, which efficiently increases susceptibility to ferroptosis ( 61 ). Blood injection results in GPX4 protein depletion in murine models with endometriosis iron overload ( 62 ). In fact, it has been reported that GPX4 expression in the early secretory phase is markedly lower in the eutopic endometrium of women with endometriosis compared with those with normal endometrium ( 63 ).…”

“…First, it is well known that the SLC3A2/SLC7A11/Nrf2/GPX4 pathway controls ferroptosis ( 60 ). It has been reported that iron administration reduced GPX4 levels in rat models (early stages of endometriosis) ( 62 ). This result suggests the importance of ferroptosis induction for cell survival during the early stages of endometriosis development.…”

“…Several lines of evidence indicate that iron has a crucial role in the proliferation of endometriotic cells. Ding et al ( 62 ) found that in comparison with control mice, endometriotic lesions in the iron-overload model mice were larger and more numerous. Under iron overload or hypoxic conditions, autophagy plays a positive role in scavenging ROS, preserving mitochondrial integrity, counteracting metabolic insults, avoiding apoptosis and protecting cells ( 10 ).…”

Double-edged roles of ferroptosis in endometriosis and endometriosis-related infertility

“…Thus, exogenous ferroptosis inducer-induced ferroptosis inhibits oocyte meiosis by increasing oxidative stress, inducing mitochondrial dysfunction, triggering autophagy splitting process and affecting oocyte quality [ 127 ]. Conversely, the inhibition of ferroptosis might not only inhibit the progression of endometriosis, but also improve the adverse effects of iron overload on ovarian function, thereby improving fertility and becoming a therapeutic approach for endometriosis-related infertility [ 128 ].…”