The ATP/P2X7 axis⁃mediated K + efflux promotes NLRP3 inflammasome activation in NDV⁃infected ECA109 cells
10.19405/j.cnki.issn1000-1492.2023.01.008
- Author:
Xu Cao
1
;
Caixia Wu
1
;
Jinping Lan
1
;
Jing Wang
1
;
Zhaoxia Jia
1
;
Hao Liu
1
;
Kaiyang Liu
1
Author Information
1. Life Science Research Center of Hebei North University,Zhangjiakou 075000
- Publication Type:Journal Article
- Keywords:
newcastle disease virus;
ATP;
P2X7;
K +;
NLRP3 inflammasome;
ECA109 cells
- From:
Acta Universitatis Medicinalis Anhui
2023;58(1):42-47
- CountryChina
- Language:Chinese
-
Abstract:
Objective :To explore whether the NLRP3 inflammasome is activated after Newcastle disease virus (NDV) exposure to esophageal cancer ECA109 cells , whether its activation is related to K + efflux , and the effect of ATP/P2X7 axis on the activation of NLRP3 inflammasome.
Methods:The expression of NLRP3 and IL⁃1β was detected by Western blot; the content of IL⁃1β in the supernatant was detected by ELISA ; the formation of ASC spots was detected by fluorescence immunoassay; the change of intracellular K + concentration was detected by fluorescent probe technology; Interventions with ATPase , ATP and P2X7 receptor inhibitors were used to investigate their role in NLRP3 inflammasome activation.
Results:Compared with the control group , the expression of NLRP3 , IL⁃1β and ASC protein in cells was up⁃regulated after NDV F3 infection ; the intracellular potassium concen tration decreased with the prolongation of infection time (P < 0. 05) . After the intervention of P2X7 receptor inhibitor, the efflux of intracellular K + was blocked. With the increase of inhibitor concentration , the efflux of K + was maximally inhibited at 10 μmol/L (P < 0. 05) . The results of ATPase and ATP intervention showed that ATPase inhibited K + efflux , while ATP promoted K + efflux. Western blot results showed that compared with the control group , P2X7 receptor was inhibited , and the expressions of NLRP3 and IL⁃1β were down⁃regulated ; after ATPase intervened cells , the expressions of NLRP3 and IL⁃1β decreased ; After ATP intervention in cells , the protein expressions of NLRP3 and IL⁃1β were up⁃regulated (P < 0. 05) .
Conclusion:NDV F3 infection of ECA109 cells can activate the NLRP3 inflammasome , the mechanism may be related to the ATP/P2X7 axis.
- Full text:2024070420484838423ATP_P2X7轴介导的K...胞中NLRP3炎性小体激活_曹旭.pdf
