ISSN 1006-298X      CN 32-1425/R

Chinese Journal of Nephrology, Dialysis & Transplantation ›› 2025, Vol. 34 ›› Issue (1): 38-44.DOI: 10.3969/j.issn.1006-298X.2025.01.007

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Acteoside  on  lipopolysaccharide-induced  human  glomerular  endothelial  cell  injury  by  regulating  the  IRE1α- TXNIP-NLRP3 signaling pathway

  

  • Online:2025-02-28 Published:2025-03-15

Abstract: Objective:To investigate the effect of acteoside (Act) on lipopolysaccharide (LPS)-induced injury to glomerular endothelial cells (GEC)by regulating the inositol-requiring enzyme 1α (IRE1α)-thioredoxin-interacting protein (TXNIP)-NOD-like receptors family pyrin domain containing 3  (NLRP3) signaling  axis.    Methodology:RT  qPCR  and Western blot experiments were used to detect the expression levels of IRE1 α mRNA and protein in transfected cells.  The human renal GEC (HRGEC) cells were separated into control (Ctrl) group, LPS group, low-dose Act group (Act-L, 50 μmol/L), high-dose Act group (Act-H, 100 μmol/L), Act-H+pcDNA-NC group, and Act-H+pcDNA-IRE1α group. The reagent kit  was  used  to  detect  the  production  of  reactive  oxygen  species  (ROS) in  HRGEC  cells.  CCK-8  method  was  applied to detect the survival of HRGEC cells. Flow cytometry was applied to detect apoptosis in HRGEC cells. ELISA assay was applied to measure the levels of IL-1β, IL-6, IL-18 and TNF-α. TNF-α in HRGEC cells. Western blot was applied to determine the  expression  levels  of  IRE1α,  p-IRE1α,  TXNIP,  and  NLRP3  proteins.     Results: compared  with  the  Ctrl group, the ROS production, apoptosis rate, the levels of inflammatory molecules IL-1β, IL-6, IL-18 and TNF-α, and the expression levels  of  p-IRE1α/IRE1α,  TXNIP,  and  NLRP3  proteins  in  HRGEC  cells  in  the  LPS  group  were  obviously higher (P<0.05), and the cell survival rate was obviously lower (P<0.05). And then, compared with the LPS group, the ROS production, apoptosis  rate,  the  levels  of  IL-1β,  IL-6,  IL-18  and  TNF-α,  and  the  expression  levels  of  p-IRE1α/IRE1α, TXNIP, and NLRP3 proteins  in  HRGEC  cells  in  the  Act-L  group, Act-H  group, and  Act-H+pcDNA-NC  group were obviously lower (P<0.05), and the cell survival rate was obviously higher (P<0.05). In the end, compared with the Act-H+pcDNA-NC  group,  the  ROS  production,  apoptosis  rate,  the  levels  of  IL-1β,  IL-6,  IL-18  and  TNF-α,  and  the expression  levels  of  p-IRE1α/IRE1α,  TXNIP,  and  NLRP3  proteins  in  the  Act-H+pcDNA-IRE1α  group  were  obviously higher (P < 0.05),  while  the  cell  survival  rate  was  obviously  lower  (P < 0.05).     Conclusion: Act  may  alleviate  LPS- induced GEC injury, promote cell growth, inhibit inflammatory molecule secretion and cell apoptosis by downregulating the expression of key proteins in the IRE1α-TXNIP-NLRP3 signaling pathway.

Key words: acteoside , inositol-requiring , enzyme , 1α-thioredoxin-interacting , protein-nod-like , receptors , family , pyrin domain containing 3 signaling axis, glomerular endothelial cells