Abstract:

Objective

To investigate the repairing effect of low-intensity ultrasound (LIPUS)on cisplatin-induced ovarian cell damage in mice.

Methods

Forty ICR mice were divided into a control group (n=12), a cisplatin group (n=14), and a cisplatin+LIPUS group (n=14).Control mice were injected intraperitoneally with saline, mice in the cisplatin group were injected intraperitoneally with cisplatin, and the cisplatin+LIPUS group underwent intraperitoneal cisplatin injection, followed by LIPUS irradiation on the next day.The treatment cycle was 2 weeks long.Mouse ovary tissues were taken for HE staining for follicle counting.RNA sequencing, immunohistochemistry, TUNEL staining, protein immunoblotting (Western blot), and real-time fluorescence quantitative polymerase chain reaction (qPCR) were performed to detect the expression of cytokines in different cellular physiological events in the ovaries such as apoptosis, iron death,and oxidative stress.

Results

Compared to the control group, ovarian size was reduced in the cisplatin group; compared to the cisplatin group, ovary size was increased in the cisplatin+LIPUS group.Comparison of ovarian weights in the control, cisplatin, and cisplatin+LIPUS groups (8.8±1.34 vs 2.8±0.45 vs 5.6±1.35)revealed statistically significant differences (P＜0.05).Compared to the control group, the number of primordial follicles, primary follicles, secondary follicles and antral follicles was reduced in the cisplatin group; compared with the cisplatin group, the number of follicles at the corresponding levels was increased in the cisplatin+LIPUS group (P＜0.05).Between mice with LIPUS treatment and controls, differential genes were mainly enriched in events at the cellular physiological level, such as apoptosis, iron death, and oxidative stress.Compared to the control group, Bax, p-H2AX, TUNEL signal, 3-Nit, 4-HNE, and 8-OHdG levels were increased, while the levels of factors such as Bcl2, SIRT1, SIRT2, Nrf2, FPN, GPX4, Fth, and Ftl were reduced in the cisplatin group (P＜0.05); compared with the cisplatin group, the levels of Bax,p-H2AX, TUNEL signal, 3-Nit, 4-HNE, and 8-OHdG were reduced, while the levels of Bcl2 and iron death factors except SIRT2 were increased (P＜0.05).

Conclusion

LIPUS can alleviate cisplatin-induced ovarian damage in mice, possibly by regulating apoptosis, iron death, and oxidative stress.

Key words: Low-intensity ultrasound, Ovary, Cisplatin, Apoptosis, Iron death, Oxidative stress, Mice