Ziyuglycoside I attenuates Deinagkistrodon acutus venom–induced injury in renal tubular epithelial cells by modulating PAR1–NLRP3–IL-1β signaling

Introduction Sanguisorba officinalis L. (Di Yu) has traditionally been used for snakebite treatment. Ziyuglycoside I, a major bioactive constituent of Di Yu, exhibits anti-inflammatory activity, but its role in snake venom-associated renal injury remains unclear. This study aimed to investigate its potential protective effects against Deinagkistrodon acutus venom (DAV)-induced renal tubular epithelial cell injury in vitro.Methods HK-2 cells were exposed to DAV to establish an in vitro renal tubular epithelial injury model. Cellular metabolic activity and intracellular ATP levels were assessed using Cell Counting Kit-8 and CellTiter-Glo assays, respectively. DAV-induced molecular alterations were examined by transcriptomic sequencing, bioinformatic analysis, western blotting, and qRT-PCR. PAR1 knockdown and pharmacological inhibition with vorapaxar were performed to determine the role of PAR1. Ziyuglycoside I was then applied to evaluate its effects on DAV-induced injury, PAR1 expression, and PAR1 ubiquitination.Results DAV dose-dependently reduced cellular metabolic activity and intracellular ATP levels in HK-2 cells, accompanied by inflammatory and apoptosis-related responses, altered cellular energy metabolism, increased NLRP3 and IL-1β expression, and decreased Bcl-2 expression. PAR1 knockdown or inhibition attenuated DAV-induced inflammatory activation and cellular injury. Ziyuglycoside I alleviated DAV-induced HK-2 cell injury, reduced PAR1 and IL-1β protein expression, and enhanced PAR1 ubiquitination, with evidence supporting proteasome-associated regulation of PAR1 protein levels.Conclusion Ziyuglycoside I may exert an in vitro protective effect against DAV-induced HK-2 cell injury, possibly by enhancing PAR1 ubiquitination and reducing PAR1 protein levels through a proteasome-associated mechanism, thereby suppressing PAR1-NLRP3-IL-1β-associated inflammatory responses.