Effects Of Potassium Channel Subtype Kv1.3and Kv1.5on Methamphetamine Induced Microglial Cell Damage

Objective To investigate the effect of methamphetamine (Meth) on primary culturedmicroglial cells in fetal rat. Methods Cell counting kit (CCK-8and MTT) andterminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) were used to detect cell viability and apoptosis by microglial cell dissected fromfetal rat. Results MTT assay shows that Meth reduces cell viability in adose-dependent manner.10,30,100,300and1000μM Meth decreased the cellviability, of all,300and1000μM Meth decreased viability significantly in statistics.CCK-8kit assay indicates that damage induced by Meth can be partially reversed byPre-incubated with TEA,4-AP (potassium channel spectrum inhibitor) andMargatoxin (MgTx, potassium channel subtypes Kv1.3specific inhibitor). TUNELassay demonstrates100and300μM Meth increase cell apoptosis, while300μMMeth increases apoptosis significantly in statistics. Conclusion Meth causesmicroglial cell damage, which could be partially reversed by potassium ion channelinhibitors TEA,4-AP and MgTx. Objective To investigate the effect of potassium Channel Subtype Kv1.3and Kv1.5on microglial cell damage induced by Meth. Method Western-blot and real-timefluorescence quantitative polymerase chain reaction (Real-time PCR) were used todetect the Kv1.3, Kv1.5mRNA expression in microglial cells. We also observe theinflammatory cytokines triggered by Meth, such as IL-6、TNF-α、iNOs. Western-blotwas applied to observe the expression of Kv1.3and p38-mitogen-activated proteinkinase (p38-MAPK) signaling pathway mediated by Meth. Results MgTxsignificantly protected against the cell damage mediated by Meth, Kv1.3rather thanKv1.5mRNA and protein expression were substantially up-regulated induced byMeth. Meth activates p38MAPK pathway, the damage induced by Meth could beblocked by Kv1.3inhibitor MgTx. Conclusion Meth activates p38MAPK pathwayand Kv1.3was involved in regulating microglial cell damage induced by Meth,Kv1.3was suggested a potential target for the development of therapeutic strategies forMeth abuse.