Transcriptomics reveals the effect of butanol extract of Pulsatilla decoction in alleviating vulvovaginal candidiasis by inhibiting neutrophil chemotaxis and activation via TLR4 signaling

Pulsatilla decoction is a renowned herbal remedy utilized in clinical settings for treating vulvovaginal candidiasis. Notably, the n-butanol extract of Pulsatilla decoction has demonstrated superior therapeutic efficacy against VVC. However, the precise mechanism underlying its effectiveness remains elusive. Transcriptomic analysis has revealed a close association between BEPD's effects and the activation and chemotaxis of neutrophils. Hence, this study aimed to elucidate the impact of BEPD on neutrophils and the relevant signaling pathways. High-performance liquid chromatography was employed to identify BEPD's primary active constituents. A VVC mouse model, induced through an estrogen-dependent method, was utilized to assess BEPD's efficacy in alleviating VVC symptoms. Comprehensive evaluations included fungal load and morphology in the vaginal cavity, alongside pathological and cytokine assays of vaginal tissues. Western blotting and immunohistochemistry were employed to analyze the expression of pertinent proteins, including CXCL1, CXCL3, S100A8, S100A9, TLR4, MyD88, myeloperoxidase, and neutrophil elastase. Immunofluorescence confirmed the colocalization of Ly6G with TLR4 and MyD88. Results indicate that BEPD treatment mitigated VVC symptoms by reducing fungal burden, inflammation, and neutrophil chemotaxis, while inhibiting the TLR4 signaling pathway, as evidenced by transcriptomics. This study highlights BEPD's pharmacological mechanism and potential for VCC treatment.