Genistein administration increases the level of superoxide dismutase and glutathione peroxidase in the endometriosis mice model: An experimental study

Abstract Background Endometriosis and infertility are caused by reactive oxygen species or free radicals, which promote endometrial cell growth and adhesion in the peritoneal cavity. Genistein has been proven to protect cells against reactive oxygen species by scavenging free radicals and decreasing the expression of genes-associated stress responses. Objective This study was conducted to determine whether genistein also acts as an antioxidant by elevating superoxide dismutase (SOD) and glutathione peroxidase (GPx) in the peritoneal fluid of the endometriosis mice model. Materials and Methods This experimental study involved 32 healthy female mice (Mus musculus), aged between 2-3 months and weighing 20-30 gr. They were divided into negative control group (healthy mice without genistein), endometriosis group (endometriosis mice without genistein), treatment group that was given different doses of genistein, that is, 0.13; 0.26; 0.52; 0.78; 1.04; and 1.3 mg/day (n = 4/each). SOD level in the peritoneal fluid was measured using the quantitative colorimetric determination method, and a colorimetric assay measured the GPx levels. Results Results showed that the endometriosis model has lower SOD and GPx levels than the control group. The administration of genistein significantly normalized these changes. Genistein significantly increased SOD levels in the 0.13 mg and 0.26 mg treatment groups. Genistein also increased GPx levels significantly in all treatment groups. Conclusion Genistein increases SOD and GPx levels in the peritoneal fluid of an endometriosis mice model, and the change is dose-dependent.


Introduction
Endometriosis is defined as a chronic gynecologic disorder in females marked by endometrial glands and stroma in the pelvic peritoneal cavity and is related to pelvic pain and infertility (1,2). Asymptomatic endometriosis occurs in 1-7% of women seeking elective sterilization, 60% among reproductive-aged women with pelvic pain, and 50-60% of women and adolescents with pelvic pain and/or unexplained infertility. The overall prevalence of endometriosis in reproductive-aged women is around 10% (1). The female reproductive system is sensitive to the damaging effects of reactive oxygen species (ROS) (3). The imbalance between oxidant generation and endogenous antioxidants is an important factor in endometriosis (3,4).
When the balance between antioxidant and ROS production gets disrupted, oxidative stress occurs as an imbalance in this ratio due to increased levels of ROS and/or reactive nitrogen species or decreased antioxidant functions (5,6).
Endometriosis is a multifaceted disease caused by complex interactions between factors such as genetic mutations, hormonal, epigenetic changes, chromosomal imbalances, and environmental risk factors such as chronic inflammation and oxidative stress, which have been implicated (4). The local pelvic inflammatory response increases the levels of cytokines, growth factors, and other anti-inflammatory mediators. Local oxidative stress in the peritoneal environment can be part of several events leading to endometriosis (2).
At present, there are no drugs that can completely cure endometriosis, and in certain situations, long-term usage might result in adverse effects. Therefore, ideal treatments are currently being developed to reduce the disease and its symptoms while avoiding the adverse effects related to hypoestrogenic conditions (7).
Mice are the most often used animal models for studying the pathophysiology of endometriosis because they are helpful and dependable tools for testing new treatment approaches in human endometriosis (8). Genistein is a natural phytoestrogen isoflavone found mainly in legumes, especially soybeans, and is an essential source of xenoestrogen (9,10). Genistein can bind to the estrogen receptor (ER) and exhibit mild estrogenic and/or antiestrogenic actions since its molecular structure is similar to that of estradiol (9). Menopausal symptoms and other endocrine illnesses have been proven to be alleviated with genistein (10).
SOD is an enzymatic antioxidant present in the body that acts as the body's first line against ROS by inactivating ROS with the superoxide-free radical O 2 − . It can protect cells and extracellular components from cell damage (11). The enzyme GPx is a significant peroxide scavenging enzyme that removes superoxide radicals, and hydrogen peroxide defends the cells from oxidative stress due to cell membrane peroxidation (2). It can prevent the formation of highly toxic hydroxyl radicals. GPx also reduces lipid or non-lipid hydroperoxides in glutathione oxidation (2,12).
Inadequate removal of superoxide and hydroxyl radicals by SOD will result in oxidative stress (13,14). Lipid peroxidation will occur in cell membranes if free radicals are not neutralized by endogenous or exogenous antioxidant molecules such as SOD and GPx (15).
Many biomarkers of oxidative stress have been studied in endometriosis with mixed results.   (17,18) and has been converted to a mice dose (19).

Collection of peritoneal tissues
Twenty-four hr after the last administration of genistein (i.e., day 30), mice were sacrificed using an anesthetic agent and dissected to collect peritoneal tissues.

Ethical considerations
The

Statistical analysis
The level of SOD

Discussion
The findings exhibited that SOD and GPx Genistein is the most abundant isoflavone in legumes with high phytoestrogen activity (30,33

Conclusion
It is concluded that genistein has the potential