The effect of silencing TRPM7 on the inflammatory response of epileptic Neuro-2a cells induced by magnesium free extracellular fluid

Objective To observe the effect of transient receptor potential M7 channel (TRPM7) gene silencing on the inflammatory response in epileptic Neuro-2a cells induced by magnesium free extracellular fluid. Methods The mouse brain neuroma cells (Neuro-2a) were transfected by TRPM7 siRNA through liposomes. Cells were cultured in magnesium free extracellular medium for 3 hours to establish an epilepsy cell model. The experiment was divided into four groups:control group (Control),epilepsy group (EP),epilepsy transfection group (EP+si-TRPM7),and epilepsy transfection negative control group (EP+si-NC). The expression of TRPM7 mRNA was detected by real-time fluorescence quantitative PCR technology 24 hours after EP model was built. The changes of HMGB1 and TLR4 protein were detected by Western Blot. ELISA measures were used to check the expression of TNF-α,IL-1β and IL-6 in the cell supernatant. CCK-8 method was used to detect the cell viability. Results Compared with the Control group,the expression of TRPM7 mRNA,HMGB1 and TLR4 protein in cells increased in EP group 24 hours after EP model was built. The expression of TNF-α,IL-1β and IL-6 in the cell supernatant increased too. While the cell viability decreased. Compared with the EP+si-NC group,the expression of TRPM7 mRNA,HMGB1 and TLR4 protein in cells decreased in EP+si-TRPM7 group. The expression of TNF-α,IL-1β and IL-6 in the cell supernatant decreased too. While the cell viability increased (P< 0.01). Conclusions Silencing TRPM7 can inhibit the HMGB1/TLR4 pathway,alleviate inflammatory response,and have a protective effect on epileptic cells induced by magnesium free extracellular fluid.