叔丁基对苯二酚对锰致神经细胞毒性的保护作用

摘要

目的 研究叔丁基对苯二酚(tBHQ)对锰致大鼠肾上腺嗜铬细胞瘤(PC12)细胞毒性的保护作用.方法 以不同终浓度(300、600、900μmol/L)的MnCl2分别处理PC12细胞24、48、72 h,用噻唑蓝(MTT)法检测PC12细胞毒性.MnCl2组予600μmol/L MnCl2处理72 h;tBHQ组予40 μmol/L tBHQ处理细胞84h;tBHQ+MnCl2组予40μmol/L tBHQ预处理12h后,600 μmol/LMnCl2处理72 h,用MTT法检测细胞毒性,MnCl2处理剂量为300 μmol/L时用Annexin V-FITC/PI法流式细胞仪(FCM)检测细胞凋亡.结果 与对照组比较,300、600、900μmol/LMnCl2处理24、48、72h,细胞增殖相对比值降低,差异均有统计学意义(P＜0.05,P＜0.01);300、600、900 μmol/L MnCl2处理组各组间相互比较,有剂量一效应关系,差异均有统计学意义(P＜0.01).与对照组比较,600 μmol/L MnCl2组抑制PC12细胞增殖,抑制率为40%,差异有统计学意义(P＜0.01).与对照组及MnCl2组比较,tBHQ组和tBHQ+MnCl2组的细胞出现增殖效应,细胞增殖相对比为1.8,差异均有统计学意义(P＜0.01).300μmol/L MnCl2处理组PC12细胞凋亡率明显高于对照组,差异有统计学意义(P＜0.01),tBHQ+MnCl2组细胞凋亡率低于300μmol/L MnCl2处理组,差异有统计学意义(P＜0.01),细胞凋亡抑制率61%.结论 锰可抑制PC12细胞增殖作用,可诱导细胞凋亡;tBHQ可减弱锰抑制PC12细胞的增殖作用并可削弱锰致PC12细胞凋亡的作用.%Objective To investigate the protective effect of the tert-butylhydroquinone (tBHQ) on PC12 cells from neurotoxicity induced by manganese.Methods Cytoyoxicity of PC12 cells was measured by MTT assay,following the PC12 cells treatment with different concentrations of MnC12 ( 300,600,900 μmol/L) for 24,48 or 72 h.PC12 cells were pretreated with 40 μmol/L tBHQ for 12 h,followed by the treatment of 600 μmol/L or 300 μmol/L MnCl2 for 72 h.Cytotoxicity of PC12 cells was measured by MTT assay,and cell apoptosis was examined by the method of Annexin V-FITC/PI in flow cytometry(FCM).Results The proliferation of PC12 cells treated with 300,600,900 μmol/L MnCl2 was suppressed in the dose dependent pattern (P＜0.01 ).Proliferation of PC12 cells treated with 600 μmol/L MnCl2 was suppressed to 40% of that in control group (P＜0.01),but the proliferation rate of PC 12 cell pretreated with 40 μmol/L tBHQ was 180% of that in control group (P＜0.01 ).Apoptotic rate of PC 12 cells treated with 300 μ mol/L MnCl2 was higher than the vehicle control group (P＜0.01).Apoptotic rate of 40 μmol/L tBHQ pretreatment followed by 300 μmol/L MnCl2 treatmcnt was lower than that of MnCl2 treatment group (P＜0.01).The inhibition rate of apoptosis was 61%.Conclusions Manganese may suppress PC12 cells proliferation and induce apoptosis,tBHQ can reduce PC12 cells proliferation suppressed by manganese and attenuate the apoptosis induced by manganese.