Study on the Inhibition of Esophageal Carcinoma Development in Mice by Salvia Chinensia Benth Induced Ferroptosis

Background Esophageal carcinoma is a common malignant tumor of the gastrointestinal tract in China and even globally, with a high incidence and mortality rate. As a traditional Chinese medicine, Salvia chinensia Benth (SJC) has the effects of clearing heat and detoxifying, promoting blood circulation and easing pain in the treatment of esophageal cancer. Pharmacological experimental studies have proved that SJC has anticancer properties and can effectively treat a variety of malignant tumors. Objective To explore the effect and mechanism of SJC in the inhibition of carcinoma in situ esophageal cancer development of C57 mice based on ferroptosis. Methods Ninety SPF grade C57BL/6 female mice were selected from February 2022 to February 2023 and randomly divided into Control group (n=15), simple 4-Nitroquinoline N-oxide (4NQO) -induced cancer group (4NQO group, n=25), 4NQO+ low-dose SJC group[4NQO/SJC (91 mg) group, n=25]and 4NQO+high dose SJC group[4NQO/SJC (182 mg) group, n=25]. The preparation of in situ model of esophageal cancer in C57 mice was carried out using 4NQO induction. The activities of the mice were observed, their mental state, food and water intake were recorded, and the body mass of the mice was measured and recorded at 8-week intervals. Hematoxylin-eosin (HE) staining and pathological analysis of esophageal tissue were performed after 32 weeks. The contents of Fe2+, glutathione (GSH) and malondialdehyde (MDA) in esophageal tissues were determined, and the expression levels of nuclear receptor coactivator 4 (NCOA4) and glutathione peroxidase 4 (GPX4) in esophageal tissues of mice were detected by western blot. Kaplan-Meier method was used to plot the survival curves of mice, and Breslow test was used to compare the survival curves. Results The body mass of mice in 4NQO group, 4NQO/SJC (91 mg) group and 4NQO/SJC (182 mg) group at 8, 16, 24 and 32 weeks of modeling was lower than that in Control group. The body mass of 4NQO/SJC (91 mg) group and 4NQO/SJC (182 mg) group at 32 weeks was higher than that of 4NQO group (P<0.05). The results of Breslow test showed that there was significant difference in the survival curves of mice in the four groups (χ2=9.907, P=0.019). The results of HE staining showed that esophageal epithelial tissue of mice in 4NQO group showed abnormal proliferation, disordered cell arrangement, and abnormal pathological changes such as keratinized beads. Compared with 4NQO group, the esophageal epithelial histopathological changes in 4NQO/SJC (91 mg) and 4NQO/SJC (182 mg) groups were significantly improved. Fe2+ and MDA in 4NQO, 4NQO/SJC (91 mg) and 4NQO/SJC (182 mg) groups were lower than those in Control group, and GSH was higher than that in Control group (P<0.05). Fe2+ and MDA in 4NQO/SJC (91 mg) and 4NQO/SJC (182 mg) groups were higher than those in 4NQO group, and GSH was lower than that in 4NQO group (P<0.05). Fe2+ and MDA in 4NQO/SJC (182 mg) group were higher than those in 4NQO/SJC (91 mg) group, and GSH in 4NQO/SJC (91 mg) group was lower than that in 4NQO/SJC (91 mg) group (P<0.05). NCOA4 in 4NQO group was lower than that in Control group, 4NQO/SJC (91 mg) group and 4NQO/SJC (182 mg) group, and GPX4 was higher than that in Control group, 4NQO/SJC (91 mg) group and 4NQO/SJC (182 mg) group (P<0.05). The GPX4 of 4NQO/SJC (91 mg) group and 4NQO/SJC (182 mg) group was higher than that of Control group (P<0.05) . Conclusion It is proved that SJC can interfere with the development of esophageal cancer development by a mechanism that may be related to NCOA4-mediated ferritin phagocytosis.