Gastrodin Improves BV-2 Microglia Pyroptosis by Inhibiting AKT/NLRP3 Pathway
- VernacularTitle:天麻素通过抑制AKT/NLRP3通路改善BV-2小胶质细胞焦亡
- Author:
Yong LIU
1
;
Ningning WANG
;
Chuhua ZENG
;
Yu WANG
Author Information
- Keywords: Gastrodin; Microglia; AKT/NLRP3 pathway; Pyroptosis
- From: World Science and Technology-Modernization of Traditional Chinese Medicine 2023;25(10):3238-3245
- CountryChina
- Language:Chinese
- Abstract: Objective To investigate the effects of Gastrodin on AKT/NLRP3 pathway in BV-2 microglial pyroptosis model.Methods BV-2 microglia were divided into normal control group and model group(1 μg·mL-1 LPS+10 μmol·L-1 Nigericin),Gastrodin group(10,100 μg·mL-1 Gastrodin),inhibitor control group(10 μmol·L-1 LY294002),inhibitor group(100 μg·mL-1 Gastrodin+10 μmol·L-1 LY294002),using 1 μg·mL-1 LPS combination 10 μmol·L-1 Nigericin co intervened to induce pyroptosis of BV-2 microglia.CCK-8 observed the effect of Gastrodin on cell activity,and western blot detected p-AKT,NLRP3,ASC,caspase-1,IL-1β expression,immunofluorescence detection of NLRP3,p-AKT protein expression,Real-time PCR observation of IL-1β mRNA content.Molecular docking was used to determine the binding sites of Gastrodin to p-AKT1/2/3 and NLRP3 proteins.Results Compared with the control group(0 μg·mL-1 Gastrodin),0.1-1000 μg·mL-1 Gastrodin had no significant effect on BV-2 cell activity(P>0.05).In the western blot experiment,compared with the normal group,the expression p-AKT,NLRP3,ASC,caspase-1,IL-1β in model group was significantly increased(P<0.01);Compared with the model group,the expression of p-AKT,NLRP3,ASC,caspase-1,IL-1β in Gastrodin group decreased(P<0.05 or P<0.01).In immunofluorescence,the expression of NLRP3 in model group was significantly higher than that in normal group(P<0.01);Compared with the model group,the expression of p-AKT and NLRP3 in Gastrodin group decreased significantly(P<0.01);Compared with Gastrodin group,the expression of p-AKT and NLRP3 in the inhibitor group decreased(P<0.01).In the Real-time PCR experiment,compared with the normal group,IL-1β mRNA content in model group cells increased significantly(P<0.01);Compared with the model group,IL-1β mRNA content in Gastrodin group cells decreased(P<0.01).In the LDH content test,compared with the normal group,the LDH activity in the model group cells increased significantly(P<0.01);Compared with the model group,the intervention of 100 μg·mL-1 Gastrodin can reduce the LDH activity in the pyroptosis cells(P<0.01).In molecular docking,Gastrodin had the binding ability with AKT1/2/3 and NLRP3,with the binding energy range of(-6.6)-(-6.8),and the binding mode included hydrogen bond,van der Waals force,etc.Conclusion Gastrodin can improve the pyroptosis of BV-2 microglia by inhibiting AKT/NLRP3 pathway.
