Protective effects of royal jelly on testicular tissue damage in rats treated with methotrexate: the relationship between oxidative stress and autophagy

The present study evaluated the protective effects of royal jelly (RJ) on methotrexate (MTX)-induced testicular damage in rats, focusing on oxidative stress and autophagy. Methotrexate, a folic acid analogue used in cancer and autoimmune treatments, impairs spermatogenesis via oxidative stress and apoptosis. Twenty-four male Wistar rats were randomized into four groups: Control (normal saline, ۳۵ days), MTX (۰.۳۰ mg kg-۱, gavage, three times per week, ۳۵ days), MTX + RJ (۰.۳۰ mg kg-۱ MTX + ۰.۱۰ mg kg-۱ RJ, gavage, three times per week, ۳۵ days), and RJ (۰.۱۰ mg kg-۱, gavage, three times per week, ۳۵ days). After ۳۵ days, rats were euthanized and testicular tissue was analyzed via histopathology, immunohistochemistry for LC۳-I/II expression in germ cells and qRT-PCR for mRNA expression of autophagy-related genes (Beclin-۱, Atg۷, LC۳-I). Histopathological findings revealed that MTX caused severe interstitial edema, coagulative necrosis and disrupted spermatogenesis with reduced seminiferous tubule diameter, epithelial thickness, tubular differentiation index (TDI) and spermiogenesis index. Co-administration of RJ significantly improved seminiferous tubule morphology, diameter, epithelial thickness, TDI and spermiogenesis index. Immunohistochemistry showed a significant increase in LC۳-I/II+ germ cells (spermatogonia, spermatocytes, spermatids) in the MTX group which was markedly reduced in the MTX + RJ group. Similarly, qRT-PCR analysis demonstrated elevated mRNA levels of Beclin-۱, Atg۷, and LC۳-I in the MTX group which were significantly reduced in the MTX + RJ group. These findings suggested that RJ mitigated MTX-induced testicular damage by reducing oxidative stress and autophagy, thereby, preserving spermatogenesis and testicular integrity.

کلیدواژه ها:

autophagy ، Methotrexate ، Oxidative stress ، qRT-PCR ، Rat