叔丁基对苯二酚对锰致神经细胞毒性的保护作用

Protective effect of tert-butylhydroquinone on PC12 cells from neurotoxicity induced by manganese in vitro

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、900umol／L）的MnCl2分别处理PCI2细胞24、48、72h，用噻唑蓝（MTT）法检测PC12细胞毒性。MnCl2组予600umol／LMnCl2处理72h；tBHQ组予40txmol／L tBHQ处理细胞84h；tBHQ＋MnCl2组予40umol／LtBHQ预处理12h后，600umol／L MnCl2处理72h，用MTT法检测细胞毒性，MnCl2处理剂量为300umol／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、900umol／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、900umol／LMnCl2处理组各组问相互比较，有剂量-效应关系，差异均有统计学意义（P〈0．01）。与对照组比较，600umol／LMnCl2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）。300umol／LMnCl2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300umol／LMnCl2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PCI2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 PC 12 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 umol/L） for 24, 48 or 72 h. PC12 cells were pretreated with 40 umol/L tBHQ for 12 h, followed by the treatment of 600 umol/L or 300 umol/L MnCl2 for 72 h. Cytotoxicity of PCI2 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 PC 12 cells treated with 300, 600, 900 umol/L MnC12 was suppressed in the dose dependent pattern （P〈 0.01 ）. Proliferation of PC12 cells treated with 600 umol/L MnCl2 was suppressed to 40% of that in control group （P〈0.01）, but the proliferation rate of PC12 cell pretreated with 40 umol/L tBHQ was 180% of that in control group （P〈0.01）. Apoptotic rate of PC12 cells treated with 300 umol/L MnCl2 was higher than the vehicle control group （P〈0.01）. Apoptotic rate of 40umol/L tBHQ pretreatment followed by 300umol/L MnCl2 treatment 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.