Electroacupuncture Ameliorates Cyclophosphamide-Induced Ovarian Impairment in Rats With Diminished Ovarian Reserve and is Associated With Th17/Treg-Related Immune Modulation.

Abstract

<p>BACKGROUND: Diminished ovarian reserve (DOR) is a major challenge in reproductive medicine, especially with delayed childbearing. Current treatments show limited efficacy and side effects. Electroacupuncture (EA), a multitarget nonpharmacological therapy, may protect ovarian function and regulate immune balance. OBJECTIVES: This study investigated the protective effects of EA and its potential mechanisms in cyclophosphamide (CTX)-induced DOR in rats, with a focus on apoptosis-related changes in granulosa cells and T helper 17 (Th17)/regulatory T (Treg)-related immune modulation. METHODS: A CTX-induced DOR rat model was treated with EA at CV4 (Guanyuan) and CV6 (Qihai), with sham EA and normal groups as controls. Estrous cycle, ovarian indices, serum hormones, and cytokines were assessed. Ovarian morphology, follicle counts, apoptosis, and protein expression were evaluated by histology, TUNEL, western blotting (WB), immunohistochemistry (IHC), and immunofluorescence (IF). Splenic Treg/Th17 cells were analyzed by flow cytometry, and RNA sequencing identified EA-regulated pathways. RESULTS: EA improved estrous cyclicity, ovarian morphology, and follicular development; improved follicle-associated marker expression; and reduced elevated FSH and LH levels in CTX-induced DOR rats. AMH and E2 showed upward trends after EA treatment, but these changes did not reach statistical significance. EA reduced TUNEL positivity, restored Ki67 expression, and favorably regulated the Bcl-2/Bax axis, while cleaved caspase-3 remained elevated and was not significantly altered. EA was also associated with normalization of Th17/Treg-related immune indices in the spleen, serum, and ovary, including reduced IL-6, IL-17A, IL-1β, TNF-α, and RORγt, and increased FOXP3, IL-10, and TGF-β1. Transcriptomic analysis revealed enrichment of immune-related pathways, consistent with the functional findings. CONCLUSION: EA ameliorated CTX-induced ovarian injury and was associated with improved follicular development, attenuation of apoptosis-related changes, and Th17/Treg-related immune modulation. These findings support a potential ovarian-protective and immunoregulatory role of EA in DOR, although further studies are required to verify endocrine efficacy and mechanistic causality.</p>