Interleukin-10 inhibits the activation of inflammasome and the levels of inflammation and pyroptosis after cerebral ischemia-reperfusion

QIN Ke

doi:10.13431/j.cnki.immunol.j.20240098

Abstract：Objective To investigate the effects of interleukin-10(IL-10)on inflammatory responses,inflammasome activation,and pyroptosis levels in the brain tissue of mice subjected to cerebral ischemia-reperfusion injury(CIRI).Methods A mouse CIRI model was established using the middle cerebral artery obstruction reperfusion(MCAO/R)method.Mice were divided into a sham group,a CIRI model group,and a CIRI+50 µg·kg-1 recombinant IL-10(rIL-10)group.Mouse hippocampal neuronal cell HT-22 was subjected to oxygen-glucose deprivation/reoxygenation(OGD/R)treatment and divided into a control group,an OGD/R model group,and an OGD/R+50 ng·ml-1 rIL-10 group.Neurological function of mice was assessed using the Zea-Longa scoring system;cerebral infarction volume was detected by 2,3,5-triphenyltetrazolium chloride(TTC)staining;reactive oxygen species(ROS)levels in HT-22 cells and mouse brain tissue were measured using a ROS kit.ELISA was used to detect the levels of IL-1β,IL-18 and tumor necrosis factor-α(TNF-α);cell counting kit-8(CCK-8),5-ethynyl-2'-deoxyuridine(EdU)assay and flow cytometry were used respectively to assess cell viability,proliferation and pyroptosis rates;Western blot was used to detect the protein expression levels of nuclear factor E2-related factor 2(Nrf2),heme oxygenase-1(HO-1),nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing protein 3(NLRP3),cleaved caspase-1,and the N-terminal fragment of gasdermin D(GSDMD-N).Results Compared with the CIRI model group,the CIRI+50 µg·kg-1 rIL-10 group showed significantly lower percentages of cerebral infarction volume and neurological function scores,lower levels of ROS,IL-1β,IL-18 and TNF-α in the brain tissue,higher expression levels of Nrf2 and HO-1 proteins,while lower levels of NLRP3,cleaved caspase-1 and GSDMD-N proteins(P<0.01).Compared with the OGD/R model group,the OGD/R+50 ng·ml-1 rIL-10 group exhibited significantly higher leve of cell viability and EdU-positive cell rates in HT-22 cells,along with significantly decreased pyroptosis rate,lower levels of ROS,IL-1β,IL-18 and TNF-α,higher levels of Nrf2 and HO-1 protein expression,while lower expression levels of NLRP3,cleaved caspase-1 and GSDMD-N proteins(P<0.01).Conclusion rIL-10 can reduce inflammatory responses,inflammasome activation,and pyroptosis levels in the brain tissue of mice subjected to CIRI,exerting protective effects against cerebral ischemia.The mechanism may be related to the activation of the Nrf2/HO-1 signaling pathway.