Diterpene ginkgolides meglumine injection(DGMI),a kind of Ginkgo biloba special extract injection,is now used for the treatment of ischemic stroke in convalescence.In the present study,we aimed to confirm whether DGMI...Diterpene ginkgolides meglumine injection(DGMI),a kind of Ginkgo biloba special extract injection,is now used for the treatment of ischemic stroke in convalescence.In the present study,we aimed to confirm whether DGMI could suppress inflammatory responses and apoptosis and explore the potential mechanisms underlying these effects.Cell viability and lactate dehydrogenase(LDH)release were measured by MTS and LDH assays after the cells were exposed to oxygen-glucose deprivation/reoxygenation(OGD/R).The extent of anti-apoptotic effect of DGMI was detected by flow cytometry using Annexin V-FITC/PI double staining assay kit.Pro-inflammatory cytokines,including TNF-α,IL-1β,IL-6 and IL-10,were quantified by a specific Bio-Plex ProTM Reagent Kit.Additionally,activities of TLR2/4,NF-κB p65,MAPK pathway and apoptosis-related proteins as well as cellular localization of NF-κB p65 were determined by Western blotting analysis and immunofluorescence staining,respectively.DGMI at 50μg/mL significantly increased the cell viability and decreased the secretion of IL-1β,IL-6,IL-10 and TNF-αin OGD/R-induced BV2 microglia cells.These effects were also confirmed by LDH assay and Annexin V-FITC/PI staining.Meanwhile,DGMI not only inhibited the protein expressions of TLR2,TLR4,MyD88,p-TAK1,p-IkBα,p-IKKβand Bak,but also decreased the cleaved caspase-3/caspase-3,Bax/Bcl-2 and cleaved PARP-1/PARP-1 ratio in OGD/R-induced BV2 microglia cells.Furthermore,OGD/R-enhanced p-JNK1/2 and p-p38 MAPK expressions and nuclear translocation of NF-κB p65 were also partially inhibited by DGMI.The present study showed that inflammatory responses were triggered in BV2 microglia cells activated by OGD/R,leading to the release of pro-inflammatory cytokines and apoptosis.DGMI suppressed the inflammatory response and apoptosis by regulating the TLR/MyD88/NF-κB signaling pathways and down-regulation of p-JNK1/2 and p-p38 MAPK activation.展开更多
文摘Diterpene ginkgolides meglumine injection(DGMI),a kind of Ginkgo biloba special extract injection,is now used for the treatment of ischemic stroke in convalescence.In the present study,we aimed to confirm whether DGMI could suppress inflammatory responses and apoptosis and explore the potential mechanisms underlying these effects.Cell viability and lactate dehydrogenase(LDH)release were measured by MTS and LDH assays after the cells were exposed to oxygen-glucose deprivation/reoxygenation(OGD/R).The extent of anti-apoptotic effect of DGMI was detected by flow cytometry using Annexin V-FITC/PI double staining assay kit.Pro-inflammatory cytokines,including TNF-α,IL-1β,IL-6 and IL-10,were quantified by a specific Bio-Plex ProTM Reagent Kit.Additionally,activities of TLR2/4,NF-κB p65,MAPK pathway and apoptosis-related proteins as well as cellular localization of NF-κB p65 were determined by Western blotting analysis and immunofluorescence staining,respectively.DGMI at 50μg/mL significantly increased the cell viability and decreased the secretion of IL-1β,IL-6,IL-10 and TNF-αin OGD/R-induced BV2 microglia cells.These effects were also confirmed by LDH assay and Annexin V-FITC/PI staining.Meanwhile,DGMI not only inhibited the protein expressions of TLR2,TLR4,MyD88,p-TAK1,p-IkBα,p-IKKβand Bak,but also decreased the cleaved caspase-3/caspase-3,Bax/Bcl-2 and cleaved PARP-1/PARP-1 ratio in OGD/R-induced BV2 microglia cells.Furthermore,OGD/R-enhanced p-JNK1/2 and p-p38 MAPK expressions and nuclear translocation of NF-κB p65 were also partially inhibited by DGMI.The present study showed that inflammatory responses were triggered in BV2 microglia cells activated by OGD/R,leading to the release of pro-inflammatory cytokines and apoptosis.DGMI suppressed the inflammatory response and apoptosis by regulating the TLR/MyD88/NF-κB signaling pathways and down-regulation of p-JNK1/2 and p-p38 MAPK activation.
