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

  • Toxicon. 2026 Dec 15:284:109312. doi: 10.1016/j.toxicon.2026.109312.
Yiwei Shen  1 ,  Naiqi Jiang  2 ,  Mengyun Tu  3
Affiliations
  • 1. Department of Breast Surgery, Affiliated Hangzhou First People's Hospital, School of Medicine, Westlake University, Hangzhou, Zhejiang, China. Electronic address: [email protected].
  • 2. Department of Clinical Laboratory, Hangzhou TCM Hospital Affiliated to Zhejiang Chinese Medical University, Hangzhou, Zhejiang, China. Electronic address: [email protected].
  • 3. Department of Clinical Laboratory, Hangzhou TCM Hospital Affiliated to Zhejiang Chinese Medical University, Hangzhou, Zhejiang, China. Electronic address: [email protected].
PMID: 42815856 DOI: 10.1016/j.toxicon.2026.109312
Abstract

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.

Keywords
Deinagkistrodon acutus venom; NLRP3 inflammasome; Protease-activated receptor 1; Renal tubular epithelial cells; Snake venom-associated kidney injury; Ziyuglycoside I.
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