Regulatory role of Keap1-Nrf2 signaling pathway in inflammatory response of in vitro subarachnoid hemorrhage cell model
YU Shi
HEI Yue
WU Kefu
LONG Qianfa
LIU Weiping
Abstract：Objective To investigate the role of Kelch-like epichlorohydrin associated protein 1(Keap1)/nuclear factor erythroid derived 2 related factor 2(Nrf2)signaling pathways on inflammatory response in subarachnoid hemorrhage(SAH)cell model.Methods A 25 μM oxyhemoglobin treatment was used in the neuron-microglia co-culture system for 24 hours to simulate the SAH cell model,which was further divided into four groups:Control,OxyHb,OxyHb+sh-Keap1 and OxyHb+sh-NC.Cell proliferation was measured by CCK8;ELISA was conducted to test the expressions of IL-6,TNF-α,IL-10,and TGF-β;DCFH probe was used to detect changes in ROS;the expressions of Keap1,Nrf2,BIP,IRE1α,PERK,p-PERK,and ATF6 in cells were measured by Western Blot,the levels of MDA and GSH were expressed using commercial kit respectively;real-time quantitative reverse transcription PCR(qRT-PCR)was conducted to detect mRNA levels of M1 surface marker(iNOS)and M2 surface marker(Arg1).Results Compared to the control group,cell proliferation in the OxyHb group decreased significantly due to oxidative stress and inflammation(P<0.05).Keap1 knockdown activated the Nrf2 signaling pathways,alleviating oxidative stress and inflammation(P<0.05),and significantly increased cell proliferation(P<0.05).The levels of M2 microglial polarization significantly increased in the OxyHb+sh-Keap1 group compared to the OxyHb group(P<0.05).Conclusions The Keap1-Nrf2 signaling pathway can regulate inflammation by modulating endoplasmic reticulum oxidative stress and microglial polarization in the SAH cell model,which may provide new therapeutic targets for the treatment of subarachnoid hemorrhage.
Keywords：kelch-like epichlorohydrin-related protein 1red-derived nuclear factor 2 related factor 2subarachnoid hemorrhage cell modelendoplasmic reticulum stressmicroglial polarization
Publication Date：2025-12-28
Online Publishing Date：2025-12-02(First online date of this platform, not the publication date of the document)
Pages：7( 21-27 )