Ginsenoside Rb2 Attenuates Microvascular Endothelial Stress Injury by Mediating Mitochondrial Pathway Pyroptosis via the <scp>DNA</scp> ‐ <scp>PKcs</scp> ‐ <scp>SIRT5</scp> Axis

The core pathogenic mechanism of coronary microvascular injury is endothelial damage following ischemic injury, subsequently triggering inflammatory responses and oxidative stress damage. This study investigated the interaction mechanism between SIRT5 and DNA-PKcs in regulating coronary microvascular endothelial injury and further validated the therapeutic potential in mitochondrial homeostasis dysregulation. We used single-cell sequencing analysis to identify dysregulation of mitochondrial homeostasis as a key regulatory phenotype in endothelial injury. The synergistic regulation of Ginsenoside Rb2 (GS-Rb2) on mitochondrial function in endothelial cells was confirmed by Western blot, RT-PCR, and other techniques. The interaction mechanism between SIRT5 and DNA-PKcs was finally identified in vitro and in vivo. By promoting the overexpression of SIRT5 and the downregulation of DNA-PKcs, GS-Rb2 intervention was able to restore heart function and reverse coronary microvascular damage during ischemia-reperfusion. Elevated DNA-PKcs expression can exacerbate microvascular endothelium inflammatory injury and disrupt endothelial cell mitochondrial homeostasis and redox balance stability. Endothelium pyroptosis was activated by SIRT5 loss, which led to a marked reduction in mitophagy and mitochondrial biogenesis. The aforesaid alterations were restored by transgenic therapy of SIRT5 and DNA-PKcs deletion, which also raised mitophagy levels, prevented excessive endothelial pyroptosis activation, and preserved microvascular cell homeostasis. At the cellular level, GS-Rb2 can boost autophagy, prevent the excessive release of DNA-PKcs in the nucleus, and improve endothelial cell activity following injury. Through SIRT5, GS-Rb2 can control DNA-PKc activity and restore mitochondrial homeostasis in the endothelium following hypoxic stress. A viable therapy approach for coronary microvascular damage was proposed.