氟哌啶醇对血清剥夺PC12细胞的保护作用及机制研究

Protective effects and mechanisms of haloperidol on PC12 cells with serum deprivation

目的探讨氟哌啶醇对剥夺血清诱导PC12细胞损伤的保护作用及其可能的机制。方法给予剥夺血清的PC12细胞不同浓度(0μm、0.1μm、0.3μm、1μm、3μm、10μm)氟哌啶醇处理,或者采用PI3K抑制剂LY294002(10μm)、Akt抑制剂AktVIII(5μm)、Mek抑制剂U0126(5μm)预处理阻断细胞信号通路,采用MTT活性分析检测细胞存活率。采用Western blot检测不同浓度、时间的氟哌啶醇对PC12细胞内Akt、FoxO1、FoxO3a、ERK等蛋白磷酸化水平的作用,采用上述三种信号通路阻断剂预处理后观察氟哌啶醇对上述蛋白磷酸化水平的影响。结果0.3~3μm氟哌啶醇可以增加血清剥夺诱导的PC12细胞存活率(P<0.05),LY294002和AktVIII可以降低其存活率(P<0.05)。氟哌啶醇和Akt、FoxO1、FoxO3a、ERK蛋白的磷酸化水平之间存在量效和时效关系。LY294002和AktVIII可以阻断氟哌啶醇引起的FoxO1和FoxO3a蛋白磷酸化水平的升高(P<0.05)。结论氟哌啶醇能够增加血清剥夺PC12细胞存活率,Akt及其下游FoxO1、FoxO3a蛋白的磷酸化水平,这可能通过PI3K/Akt-FoxOs信号通路发挥作用。

Objective To explore the protective effects and related mechanisms of haloperidol on serum deprivation-induced injury in PC12 cells.Methods Serum free PC12 cells were treated with haloperidol of various concentrations(0,0.1,0.3,1,3,10μm),or with kinase blockers LY294002(10μm),Akt VIII(5μm),and U0126(5μm)before the administration of haloperidol.MTT was used to detect cell viability,and Western blot was used to detect Akt,FoxOs,and ERK protein phosphorylation in PC12 cells in different concentrations of haloperidol and at different times of treatment.The effects of haloperidol on aforementioned protein phosphorylation levels in PC12 cells were tested in the condition of pretreatment with the three inhibitors of signal pathway.Results Significant increases in cell viability were observed with haloperidol 0.3~3μm treatments(P<0.05).However,the significant increase could be inhibited by the pretreatments of LY294002 and AktVIII(P<0.05).There were significant dose-effect and time-effect relationships between haloperidol and Akt,FoxO1,FoxO3a,ERK protein phosphorylation levels.LY294002 and AktVIII could block the haloperidol-induced FoxOs phosphorylation(P<0.05).Conclusion Haloperidol may increase viability of PC12 cells with serum deprivation,Akt and FoxOs phosphorylation,which may be associated with the PI3K/Akt-FoxOs signal pathway.