Glutamate treatment can lead to Ca2+overload,and the role and possible mechanism of tran-sient receptor potential vanilloid 4(TRPV4)in Ca2 overload are still unclear.β-Asarone can quickly penetrate the blood-brain barrier and has a strong neuroprotective effect on excitotoxicity.In this study,taking highly differentiated PC 12 cells as the research object,the effect of β-asarone(15,30,60 μmol/L)pretreatment for 4 h,40 mmol/L glutamate treatment on the Ca2+concentration of PC12 cells was recorded in real time,and the change of Ca2+concentration was detected by calcium imaging technology;TRPV4 mRNA and protein expression was detected by fluorescent quantitative polymer-ase chain reaction,Western Blot,and immunofluorescence techniques;Lipofectiamine 2000 liposome experiment was used to transfect TRPV4-siRNA and pEX-3-TRPV4 to observe the effect of silencing and overexpression of TRPV4 on glutamate-induced Ca2+overload.The results showed that compared with the control group,glutamate treatment for 5 min could induce Ca2+overload,and the mRNA and protein expression of TRPV4 was significantly increased;compared with the model group,β-asarone could reduce glutamate-induced Ca2+overload and TRPV4 expression in a dose-dependent manner;si-lencing of TRPV4 inhibited Ca2+overload in cells;overexpression of TRPV4 partially reversed the in-hibition of glutamate-induced Ca2+overload by β-asarone.This study demonstrated that glutamate treatment of PC12 cells for 5 min induced Ca2+overload by up-regulating the expression of TRPV4,and β-asarone,as an antagonist of TRPV4,is a potential drug to inhibit excitotoxicity.
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