PRDX1 modulates LPS-triggered inflammatory response and cell apoptosis in microglial cells through TLR4 signaling axis
Objective To investigate the roles and potential mechanism of PRDX1 on inflammatory responses and apoptosis in lipopolysaccharide(LPS)-induced BV2 cells.Methods BV2 cells were divided into control group(BV2 cells were normally cul-tured),LPS treated group,NC shRNA lentivirus infection group(cells were infected with NC shRNA lentivirus for 48 hours and then treated with LPS for 24 hours),and PRDX1 shRNA lentivirus infec-tion group(cells were infected with PRDX1 shRNA lentivirus for 48 hours and then treated with LPS for 24 hours).For the TLR4 overex-pression experiment,a TLR4 overexpression group was added on the basis of the lentivirus infection experiment(cells were infected with PRDX1 shRNA lentivirus and TLR4 overexpression lentivirus for 48 hours,and then treated with LPS for 24 hours).RT-qPCR and Western blot were adopted to estimate the mRNA and protein expres-sion,respectively.The evaluation of cell viability was executed by MTT assay.Cell apoptosis was appraised by flow cytometry.ELISA assays were implemented for the measurement of the release of inflammatory factors such as TNF-α and IL-6.Results Test results of ELISA showed that compared with BV2 cells treated with LPS,the levels of TNF-α and IL-6 in the PRDX1 shRNA lentivirus infec-tion group were significantly reduced(P<0.01).The RT qPCR results showed that the mRNA expression levels of TNF-α and IL-6 were consistent with the trend of ELISA test results.The MTT assay results showed that compared with the control group,the viability of BV2 cells in the LPS treatment group was significantly reduced;Compared with the LPS treatment group,the PRDX1 shRNA lenti-virus infection group showed a significant increase in cell viability.The results of flow cytometry analysis showed that compared with the control group,the apoptosis rate of BV2 cells in the LPS treatment group was significantly increased,while compared with the LPS treatment group,the apoptosis rate of PRDX1 shRNA lentivirus infection group was significantly reduced.Western blot results showed that compared with the control group,the expression of TLR4 was significantly upregulated in BV2 cells treated with LPS(P<0.05);Compared with BV2 cells treated with LPS,PRDX1 shRNA lentivirus infection significantly inhibited TLR4 expression induced by LPS(P<0.05).Compared with the PRDX1 shRNA lentivirus infection group,the TLR4 overexpression group showed significantly in-creased levels of inflammatory factors TNF-α and IL-6 in BV2 cells(P<0.05),decreased cell viability levels(P<0.05),and signifi-cantly increased levels of cell apoptosis(P<0.05).Conclusion Perturbation of PRDX1 visibly promoted cell viability,and restrict-ed inflammatory response and cell apoptosis process in LPS-evoked BV2 cells via positive regulating TLR4 expression.These findings elucidated the novel role of PRDX1/TLR4 axis in regulating SCI,and may be served as a promising therapeutic target for SCI treatment.
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