To investigate the effects of stress on gastric mucosal inflammation and ovarian function in female mice, and the inhibitory effect of Femoston.

8-week old female SPF Kunming mice were randomly divided into control, stress and Femoston intervention group (6 in each group); Female mice in the stress group and Femoston group were restricted in the self-made restraint apparatus for 2 hours every day for 14 days. The histopathological changes of gastric mucosa and ovary were observed under light microscope by H&E staining. The expression of Nox-4 in gastric mucosa and ovary was observed by immunohistochemistry. Further, the levels of Nox-4, antioxidant enzymes (Mn SOD, GSH Px, Catalase) and cytokines (IL1-β, IL-6, TNF-α) in mice ovarian tissue and blood were detected by real-time quantitative RT-PCR and ELISA.

H&E staining showed that neutrophils, lymphocytes infiltration and inflammatory changes in gastric mucosa of stress mice; The score of gastric mucositis in stress group was higher(P<0.01). Nox-4 was highly expressed in the gastric mucosa, and its mRNA expression level was increased (P<0.01). Stress increased the mRNA level of gastric cytokines (IL1-β, IL-6, TNF-α).The ovarian tissue of stress showed obvious changes in follicle microstructure, such as primordial follicle (pr), primary follicle (prm), secondary follicle (sf), inflammation (ii) and follicle structure deformation (fd). Femoston improved the abnormality of this follicle structure. The Nox-4 in ovarian mainly expressed in follicular cells; Its mRNA expression level increased (P<0.001); The content of Nox-4 in the blood of stress mice increased (P<0.001). Femoston can effectively reduce the high expression of Nox-4 in ovarian tissue and blood of stressed mice (P<0.01). Stress mice increased the levels of follicle stimulating hormone (FSH) and luteinizing hormone (LH) in blood; The levels of estradiol (E2) and progesterone (Prog) were decreased (P<0.001). Femoston improved the level of E2 (P<0.01). Stress mice decreased the mRNA level of antioxidant enzymes (Mn SOD, GSH Px, Catalase) in ovarian tissue(P<0.001); Femoston increased the expression levels of antioxidant enzymes(P<0.01). Stress increased in blood and ovarian tissue IL1-β, IL-6, TNF-α mRNA levlels, and Femoston effectively inhibited the high expressions of those cytokines(P<0.01). Pearson correlation analysis showed that there was a positive correlation between the serum level of Nox-4 and sex hormones (FSH, LH, E2, Prog) in the stress group (P<0.01). Serum Nox-4 and cytokines(IL1-β, IL-6, TNF-α) were a positive correlation between concentrations (P<0.01).

The high expression of Nox-4 in local tissues and blood is related to the gastric mucositis and abnormal ovarian function induced by stress. Femoston can effectively improve the abnormality of ovarian structure and function, deeper mechanism needs further exploration.