Chlorogenic Acid Attenuates Busulfan-Induced Azoospermia in Rats: Antioxidant-Mediated Restoration of Spermatogenesis and Steroidogenesis - A Stereological and Molecular Analysis
Abstract
Objective: Busulfan, a chemotherapeutic alkylating agent, induces azoospermia by generating reactive oxygen species (ROS), thereby impairing spermatogenesis and steroidogenesis, and threatening fertility in cancer patients. Chlorogenic acid (CGA), a polyphenolic compound with antioxidant and anti-inflammatory properties, may protect against such reproductive toxicity. This study evaluated the effects of CGA on testicular function in a rat model of busulfan-induced azoospermia, with emphasis on spermatogenesis, steroidogenesis, and testicular architecture.
Methods: Twenty-four adult male Wistar rats were randomly assigned to four groups: control, CGA (50 mg/kg, oral, for 60 days), busulfan (BUS, 40 mg/kg, single intraperitoneal dose), and BUS+CGA. Testicular function was assessed using stereological analyses (testis weight; seminiferous tubule and germinal epithelium volumes; germ and somatic cell counts), sperm parameters, serum hormone levels, oxidative stress markers (total antioxidant capacity [TAC] and malondialdehyde [MDA]), and expression of steroidogenic genes (STAR, CYP11A1, and HSD17B3) by qRT-PCR.
Results: Busulfan markedly reduced testis weight, sperm quality, testosterone, TAC, and expression of steroidogenic genes, while increasing MDA and sperm abnormalities (p<0.05). CGA co-administration significantly improved these parameters, restoring gene expression and TAC toward control levels, reducing MDA, and enhancing testosterone, sperm count, morphology, and…
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