Isoflurane-Assisted Electroacupuncture in a Rat Model of Oligoasthenozoospermia

Oligoasthenozoospermia is one of the most common forms of male reproductive dysfunction. In rodent studies, delivering electroacupuncture (EA) consistently across repeated sessions poses a technical challenge, particularly when combining abdominal midline and hind-limb acupoints, where stable electrode contact is critical. Zhongji (CV3), Guanyuan (CV4), Zusanli (ST36), and Sanyinjiao (SP6) were selected because these points are frequently used in acupuncture protocols for infertility and semen abnormalities, and the abdominal midline plus hind-limb configuration suits standardized electrode pairing. This report describes a practical protocol for establishing an adenine-induced rat model of oligoasthenozoospermia and for administering isoflurane-assisted EA with paired stimulation of CV3-CV4 and ST36-SP6. Male Sprague-Dawley rats received adenine by daily gavage for 28 days to induce reproductive impairment. Subsequently, EA was administered daily for 28 days under inhalational anesthesia, using 3% isoflurane for induction and 1.5% for mask maintenance, to enable stable needle placement and electrode pairing (CV3-CV4 and ST36-SP6) with defined stimulation parameters (2 Hz, 1.5 mA, 30 min). Treatment effects were evaluated via computer-assisted semen analysis for sperm count and motility, supplemented by testicular hematoxylin and eosin (HE) histology. Representative results indicate that adenine exposure reduced sperm count and motility and disrupted seminiferous tubule architecture, while EA was associated with partial recovery in both semen parameters and testicular morphology. This protocol offers a ready reference for studies requiring consistent EA delivery and semen/histological verification in a rat model of oligoasthenozoospermia.