BACKGROUND Neuropathic pain(NP)is the primary symptom of various neurological condi-tions.Patients with NP often experience mood disorders,particularly depression and anxiety,that can severely affect their normal live...BACKGROUND Neuropathic pain(NP)is the primary symptom of various neurological condi-tions.Patients with NP often experience mood disorders,particularly depression and anxiety,that can severely affect their normal lives.Microglial cells are as-sociated with NP.Excessive inflammatory responses,especially the secretion of large amounts of pro-inflammatory cytokines,ultimately lead to neuroinflam-mation.Microglial pyroptosis is a newly discovered form of inflammatory cell death associated with immune responses and inflammation-related diseases of the central nervous system.METHODS Two models,an in vitro lipopolysaccharide(LPS)-stimulated microglial cell model and a selective nerve injury model using BTX-A and SPP1 knockdown treatments,were used.Key proteins in the pyroptosis signaling pathway,NLRP3-GSDMD,were assessed using western blotting,real-time quantitative polymerase chain reaction,and immunofluorescence.Inflammatory factors[interleukin(IL)-6,IL-1β,and tumor necrosis factor(TNF)-α]were assessed using enzyme-linked immuno-sorbent assay.We also evaluated microglial cell proliferation and apoptosis.Furthermore,we measured pain sensation by assessing the delayed hind paw withdrawal latency using thermal stimulation.RESULTS The expression levels of ACS and GSDMD-N and the mRNA expression of TNF-α,IL-6,and IL-1βwere enhanced in LPS-treated microglia.Furthermore,SPP1 expression was also induced in LPS-treated microglia.Notably,BTX-A inhibited SPP1 mRNA and protein expression in the LPS-treated microglia.Additionally,depletion of SPP1 or BTX-A inhibited cell viability and induced apoptosis in LPS-treated microglia,whereas co-treatment with BTX-A enhanced the effect of SPP1 short hairpin(sh)RNA in LPS-treated microglia.Finally,SPP1 depletion or BTX-A treatment reduced the levels of GSDMD-N,NLPRP3,and ASC and suppressed the production of inflammatory factors.CONCLUSION Notably,BTX-A therapy and SPP1 shRNA enhance microglial proliferation and apoptosis and inhibit microglial death.It improves pain perception and inhibits microglial activation in rats with selective nerve pain.展开更多
Hepatocellular carcinoma(HCC)is associated with poor prognosis and fluctuations in immune status.Although studies have found that secreted phosphoprotein 1(SPP1)is involved in HCC progression,its independent prognosti...Hepatocellular carcinoma(HCC)is associated with poor prognosis and fluctuations in immune status.Although studies have found that secreted phosphoprotein 1(SPP1)is involved in HCC progression,its independent prognostic value and immune-mediated role remain unclear.Using The Cancer Genome Atlas and Gene Expression Omnibus data,we found that low expression of SPP1 is significantly associated with improved survival of HCC patients and that SPP1 expression is correlated with clinical characteristics.Univariate and multivariate Cox regression confirmed that SPP1 is an independent prognostic factor of HCC.Subsequently,we found that T cell CD4 memory-activated monocytes,M0 macrophages,and resting mast cells showed significant differences in penetration in the high and low SPP1 expression groups.Next,we used the Weighted Gene Co-Expression Network and Least Absolute Shrinkage Sum Selection Operator algorithms to construct a risk score for the 9-immune-related genes signature.The risk score showed a good ability to identify high and low-risk patients and improved survival prediction.We also used multivariate Cox regression to validate that risk score was significantly correlated with SPP1 and overall survival.Lastly,the Back-Propagation Neural Network confirmed the reliability of the results of multiple algorithms.In conclusion,the findings suggest that SPP1 is an independent marker of HCC survival and immunotherapy.展开更多
文摘BACKGROUND Neuropathic pain(NP)is the primary symptom of various neurological condi-tions.Patients with NP often experience mood disorders,particularly depression and anxiety,that can severely affect their normal lives.Microglial cells are as-sociated with NP.Excessive inflammatory responses,especially the secretion of large amounts of pro-inflammatory cytokines,ultimately lead to neuroinflam-mation.Microglial pyroptosis is a newly discovered form of inflammatory cell death associated with immune responses and inflammation-related diseases of the central nervous system.METHODS Two models,an in vitro lipopolysaccharide(LPS)-stimulated microglial cell model and a selective nerve injury model using BTX-A and SPP1 knockdown treatments,were used.Key proteins in the pyroptosis signaling pathway,NLRP3-GSDMD,were assessed using western blotting,real-time quantitative polymerase chain reaction,and immunofluorescence.Inflammatory factors[interleukin(IL)-6,IL-1β,and tumor necrosis factor(TNF)-α]were assessed using enzyme-linked immuno-sorbent assay.We also evaluated microglial cell proliferation and apoptosis.Furthermore,we measured pain sensation by assessing the delayed hind paw withdrawal latency using thermal stimulation.RESULTS The expression levels of ACS and GSDMD-N and the mRNA expression of TNF-α,IL-6,and IL-1βwere enhanced in LPS-treated microglia.Furthermore,SPP1 expression was also induced in LPS-treated microglia.Notably,BTX-A inhibited SPP1 mRNA and protein expression in the LPS-treated microglia.Additionally,depletion of SPP1 or BTX-A inhibited cell viability and induced apoptosis in LPS-treated microglia,whereas co-treatment with BTX-A enhanced the effect of SPP1 short hairpin(sh)RNA in LPS-treated microglia.Finally,SPP1 depletion or BTX-A treatment reduced the levels of GSDMD-N,NLPRP3,and ASC and suppressed the production of inflammatory factors.CONCLUSION Notably,BTX-A therapy and SPP1 shRNA enhance microglial proliferation and apoptosis and inhibit microglial death.It improves pain perception and inhibits microglial activation in rats with selective nerve pain.
基金supported by Hubei Provincial Key Laboratory Opening Project of Renmin Hospital of Wuhan University(2022KFH010).
文摘Hepatocellular carcinoma(HCC)is associated with poor prognosis and fluctuations in immune status.Although studies have found that secreted phosphoprotein 1(SPP1)is involved in HCC progression,its independent prognostic value and immune-mediated role remain unclear.Using The Cancer Genome Atlas and Gene Expression Omnibus data,we found that low expression of SPP1 is significantly associated with improved survival of HCC patients and that SPP1 expression is correlated with clinical characteristics.Univariate and multivariate Cox regression confirmed that SPP1 is an independent prognostic factor of HCC.Subsequently,we found that T cell CD4 memory-activated monocytes,M0 macrophages,and resting mast cells showed significant differences in penetration in the high and low SPP1 expression groups.Next,we used the Weighted Gene Co-Expression Network and Least Absolute Shrinkage Sum Selection Operator algorithms to construct a risk score for the 9-immune-related genes signature.The risk score showed a good ability to identify high and low-risk patients and improved survival prediction.We also used multivariate Cox regression to validate that risk score was significantly correlated with SPP1 and overall survival.Lastly,the Back-Propagation Neural Network confirmed the reliability of the results of multiple algorithms.In conclusion,the findings suggest that SPP1 is an independent marker of HCC survival and immunotherapy.
