# Determination of Effect of Morus alba Extract and Lactobacillus rhamnosus on Vaginal Microbiota and SAP Gene (1–10) Profile With Candida albicans Exosome Administration in Rats

## TL;DR

This study shows that combining Morus alba extract and Lactobacillus rhamnosus can reduce Candida albicans virulence and restore vaginal health in rats.

## Contribution

The novel finding is the synergistic effect of Morus alba and Lactobacillus rhamnosus in suppressing Candida virulence genes and inflammation.

## Key findings

- Combination therapy reduced all SAP1–10 gene expressions and inflammation markers like IL-6 and TLR4.
- MA reduced Mycobacteriales abundance, while LR maintained Lactobacillales levels in the vaginal microbiota.
- Histopathology showed a 60–72% reduction in ovarian inflammation with combination therapy.

## Abstract

Vaginal microbiota is essential for mucosal immunity, pathogen defense, and homeostasis. Disruption of this balance can promote opportunistic infections, notably by Candida albicans. This study investigates the therapeutic potential of Morus alba (MA) extract and Lactobacillus rhamnosus (LR) in a rat model exposed to Candida albicans exosomes (CAE). CAE induced a 1.3–5.1‐fold upregulation in SAP1–10 gene expression, with SAP4 showing the highest increase (p ≤ 0.05). MA and LR monotherapies selectively suppressed SAP6 (41%) and SAP4 (23%), respectively. Notably, combination therapy (CAE+MA+LR) synergistically inhibited all SAP genes (0.8–1.1‐fold, p ≤ 0.05). 16S rRNA analysis showed that LR‐containing groups maintained Lactobacillales abundance (37.77%–38.7%), while MA reduced Mycobacteriales by 68.5% (p = 0.004). Microbial diversity was lower in the MA group (H = 3.242) but higher in LR groups (H = 5.493–5.598). Histopathology revealed severe ovarian inflammation in the CAE group (87.4%, p = 0.0022) with a 4.2‐fold increase in IL‐6, while combination therapy reduced inflammation and IL‐6 levels by 60%–72% (p ≤ 0.05). Immunofluorescence confirmed downregulation of TLR4 and Caspase‐3 with treatment. FIB‐SEM and NTA analyses showed that CAE exosomes had a heterogeneous morphology (99.3 ± 31.1 nm) and high polydispersity (PDI = 0.31). These results suggest that MA and LR synergistically reduce fungal virulence, restore microbial balance, and offer a promising adjuvant strategy against vaginal candidiasis.

The vaginal microbiota plays a crucial role in mucosal immunity and defense against pathogens. Disruption of this balance by Candida albicans exosomes (CAE) upregulates virulence‐related SAP1–10 genes and induces inflammation. In a rat model, treatment with Morus alba (MA) extract and Lactobacillus rhamnosus (LR) demonstrated selective suppression of SAP gene expression and restoration of microbial homeostasis. Combination therapy (MA + LR) synergistically reduced SAP1–10 expression, IL‐6, TLR4, and Caspase‐3 levels, leading to decreased ovarian inflammation and reestablished Lactobacillales abundance. These results highlight the potential of MA and LR as a synergistic adjuvant approach against CAE‐induced vaginal dysbiosis.