Triptolide Alleviates Hypoxic/Reoxygenation Injury of Cortical Neurons by Inhibiting TLR4/NF-κB Pathway
Objective:To investigate the effects of Triptolide(TPL)on hypoxia/reoxygenation(H/R)injury of cortical neurons and explore its mechanism based on toll-like receptor 4/nuclear factor-κB(TLR4/NF-κB)signaling pathway.Methods:The cortical neurons of SD suckling rats were isolated and cultured in vitro,and the H/R damaged cortical neuron model was prepared by hypoxia for 4h and then reoxygenation for 24h.The normal control group,model group,TPL(25mg/L)group,TPL(25mg/L)+TAK242(TLR4 in-hibitor,1mg/L)group,TPL(25mg/L)+LPS(TLR4 agonist,0.1mg/L)group were set up.24h after ad-ministrating separately,the neuronal activity was detected by MTT,the neuronal apoptosis was detected by flow cytometry.The content of inflammatory factors(TNF-α,IL-1β,IL-6)were detected by ELISA.The TLR4/NF-κB signaling pathway related proteins[TLR4,NF-κB p65,p-NF-κB p65,B-lymphoblastoma-2 gene(bcl-2),bcl-2 associated X protein(Bax),Caspase-3,cleaved Caspase-3]were detected by West-ern blot.Results:Compared with the model group,the activity of cortical neuron in TPL group was increased while the apoptosis rate was decreased(P<0.05).The content of TNF-α,IL-1β,IL-6 in culture medium were significantly decreased(P<0.05).The expression of TLR4 and the expression ratio of p-NF-κB p65/NF-κB p65,Bax/bcl-2,cleaved Caspase-3/Caspase-3 were significantly decreased(P<0.05).TAK242 could significantly enhance the regulatory effects of TPL on the activity,apoptosis,inflammatory factors con-tent and the expression of TLR4/NF-κB signaling pathway related protein in H/R damaged cortical neurons.And LPS could significantly reversed the regulatory effects of TPL on H/R damaged cortical neurons.Conclu-sion:TPL can reduce inflammation and apoptosis by inhibiting the activation of TLR4/NF-κB pathway,and thus play a protective role in H/R injury of cortical neurons.
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