## 4 Discussion
In this study, we evaluated the inhibitory effects of the THY-F51 CFS and live cells on the production of four key inflammatory mediators (TNF-\(\alpha\), IL-1B, IL-6, and IL-8) induced by C. _albicans_ in HeLa cells. As shown in Figure 7, the inhibition rates for TNF-\(\alpha\) were found to be 49.60% for the CFS and 55.90% for live cells, while IL-1B inhibition rates were 70.06% for the CFS and 73.74% for live cells, both surpassing the positive control, Amphotericin B, which showed a 65.03% inhibition rate. For IL-6, the inhibition rates were 14.98% for the CFS and 21.62% for live cells, with live cells demonstrating a significantly stronger inhibitory effect. In the case of IL-8, the inhibition rates were 48.73% for the CFS and 66.37% for live cells, with live cells showing a higher inhibitory effect than Amphotericin B, which exhibited a 53.38% inhibition rate. These findings suggest that THY-F51 live cells exert more potent anti-inflammatory effects compared to the CFS. This difference may be due to two main mechanisms: first, live cells may interact directly with immune cells by binding to pattern recognition receptors (PRRs), such as Toll-like receptors (TLRs), on host cells, which induces the suppression of inflammatory mediators and activates intracellular signaling pathways to inhibit inflammation [41]. Second, live bacteria can modulate immune responses through the secretion of metabolites (e.g., lactic acid, acetic acid) or cell wall components (e.g., peptidoglycan, teichoic acids); although the CFS contains metabolites secreted by live cells, live cells are more effective due to their involvement in intercellular interactions and their continuous production of immune-modulating metabolites [42]. Antioxidants play a critical role in alleviating _Candida_-induced vaginitis by neutralizing excessive ROS, which contribute to oxidative stress, promote _Candida_ overgrowth, and compromise the integrity of the vaginal mucosa. These compounds suppress inflammatory pathways, such as NF-\(\kappa\)B, thereby reducing the production of pro-inflammatory cytokines and promoting the growth of lactic acid bacteria, which maintain the acidic vaginal pH required to inhibit _Candida_ proliferation [43]. Moreover, antioxidants enhance immune responses by modulating T-cell and macrophage activity and activating the Nrf2-Keap1 signaling pathway, which upregulates antioxidant-related gene expression and supports tissue regeneration. Collectively, these mechanisms mitigate infection, reduce inflammation, and promote mucosal healing [44]. Through these mechanisms, we also validated the antioxidant efficacy of THY-F51 in our study. As detailed in Table 2, the CFS of THY-F51 exhibited superior antioxidant properties compared to live cells. This is likely due to the presence of metabolic byproducts and secondary metabolites in the CFS, such as peptides, organic acids, and phenolic compounds, which are known to have strong antioxidant activity [45]. Furthermore, the ORAC value of the THY-F51 CFS was measured to be 220.45 umol TE/g, which is comparable to the ORAC value of _Lactobacillus brevis_ 603, reported as 237.96 \(\pm\) 49 umol TE/g by [PERSON] et al. [46]. Similarly, the T-AOC value of THY-F51 CFS was 5.27 U/mL, significantly higher than the reported T-AOC value of _L. plantarum_ of 0.78 U/mL [47]. These findings underscore the substantial antioxidant potential of the THY-F51 CFS and its relevance in addressing oxidative stress-related conditions, such as _Candida_-induced vaginitis. Cell adhesion inhibition plays a crucial role in preventing pathogenic microorganisms, such as _Candida_, from adhering to host cells, which helps avoid infection, biofilm formation, and associated inflammatory responses [48]. In this study, the THY-F51 CFS demonstrated a significant reduction in _Candida_ adhesion to HeLa cells, with a more substantial inhibitory effect compared to live THY-F51 cells (Figure 8a).