Tri-Culture Fermentation of Neem (Azadirachta indica) Leaves Induces Phytochemical Remodeling and Enhances Multi-Target Bioactivities Relevant to Androgenetic Alopecia
Abstract
Androgenetic alopecia (AGA) involves dihydrotestosterone-driven follicular miniaturization compounded by oxidative stress and perifollicular inflammation, while current pharmacotherapies remain limited by adverse effects. Neem (Azadirachta indica A. Juss., Meliaceae) leaves, a phenolic- and limonoid-rich plant widely used in Southeast Asian traditional medicine, were fermented for seven days with a tri-culture consortium of Saccharomyces cerevisiae, Lactobacillus plantarum, and Aspergillus niger (TRI-NE) to evaluate whether expanded microbial diversity enhances bioactivity relative to the unfermented extract (UN-NE). TRI-NE significantly increased total phenolic content, rising from 315.30 to 564.70 mg GAE/g by day 7, alongside an overall enhancement of antioxidant capacity across all assays. Moreover, untargeted metabolomics revealed compositional remodeling, with 67.47% of significantly altered metabolite features upregulated after fermentation, including selective enrichment of quercetin despite reductions in other polyphenols. In human hair follicle dermal papilla cells (HFDPCs) and complementary models, TRI-NE consistently outperformed UN-NE, enhancing paracrine-mediated fibroblast proliferation, preserving cell viability under potassium-channel blockade, suppressing lipopolysaccharide-induced inflammatory nitric oxide production, and attenuating oxidative membrane damage. At the transcriptional level, TRI-NE downregulated androgen metabolism (SRD5A1 and SRD5A2) and…
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