关键词: 毛蕊花糖苷, 肌醇需要酶1α-硫氧还蛋白相互作用蛋白-Nod 样受体蛋白3 信号轴, 肾小球血管内皮细胞

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