## 1 Introduction  
Bacterial persistence is a significant challenge in the treatment of chronic and recurrent infections caused by _Staphylococcus aureus_ due to its ability to form persister cells [1]. Unlike genetically resistant strains, _S. aureus_ cells exhibit transient antibiotic tolerance while in this metabolically dormant state, allowing them to evade antibiotic treatments without genetic mutation [2]. This reversible state is a critical survival strategy that enables persister cells to withstand bactericidal agents, breaking dormancy after antibiotic exposure, which contributes to infection relapse and prolonged treatment courses [3]. The clinical significance of persister cells is particularly pronounced in infections such as endocarditis, osteomyelitisitis, and cystic fibrosis, where repeated antibiotic treatment often fails to eradicate the entire bacterial population, resulting in persistent, refractory infections [1]. Existing therapeutic strategies for combating persister cells primarily focus on eradication through mechanisms that disrupt membrane integrity, interfere with metabolic processes, or induce oxidative stress [4; 5]. Compounds like membrane-permeabilizing agents [6; 7], DNA cross-linkers [8], and protease activators [9] have been investigated to target and eliminate preformed persister cells. However, these approaches have notable limitations. They generally fail to prevent the initial formation of persister cells and may induce collateral damage to non-persister bacterial populations or host tissues, thereby limiting their clinical applicability [10; 11]. Moreover, most current strategies overlook the metabolic pathways involved in persister cell formation, focusing solely on killing dormant cells rather than preventing their development. Addressing this gap, targeting the stringent response, a well-established regulatory pathway in bacterial persistence, has emerged as a promising approach [12]. The stringent response is mediated by the alarmone (p)ppGpp, a signaling molecule that downregulates cellular metabolism and promotes dormancy in response to nutrient deprivation, antibiotic stress, and other environmental cues [13]. In _S. aureus_, the synthesis of (p)ppGpp is regulated by the _relP_ and _relQ_ genes, both of which encode small (p)ppGpp synthases [14]. Previous studies have demonstrated that the genetic knockout of these genes significantly reduces persister cell formation, suggesting that inhibiting (p)ppGpp synthesis could effectively prevent persister cell establishment [15]. Despite this approach's therapeutic potential, few compounds have been identified that specifically inhibit (p)ppGpp synthesis without inducing cytotoxic effects or compromising bacterial viability [16; 17]. Natural compounds have recently gained attention as potential anti-persister agents due to their diverse pharmacological profiles and low toxicity compared to synthetic drugs [18; 19; 20]. Diosogenin, a naturally occurring steroidal saponin found in various plant species, is a notable example [21]. Structurally analogous to cholesterol, diosogenin can integrate into bacterial membranes, potentially affecting membrane fluidity, intracellular signaling, and metabolic pathways [22]. It has been extensively studied for its anti-cancer [23], anti-inflammatory [24], and anti-bacterial biofilm activities [25]. In the context of bacterial infections, diosogenin has been reported to inhibit biofilm formation in _S. aureus_ by regulating sigma factor B (\(\sigma^{\text{B}}\)), a key factor involved in stress response and biofilm development [26]. Despite its known biological functions, the details of diosogenin's effects on persister cell formation and (p)ppGpp synthesis remain underexplored. Given its structural similarity to cholesterol, diosogenin may disrupt membrane potential and affect intracellular signaling pathways.