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目的 :研究维拉帕米对甲基苯丙胺 (MA)导致神经细胞损伤的影响 ,探讨维拉帕米的保护机制。方法 :采用MTT法 ,乳酸脱氢酶 (LDH)法 ,观察MA的神经毒性作用。运用DNA电泳法、流式细胞仪法 ,观察MA致神经细胞凋亡的作用 ,并研究了维拉帕米的保护作用。结果 :(1 )MA(0 .2 5～ 3 0 0mmol/L)作用 4 8h和 6 0h ,能增加大鼠小脑神经细胞系R2细胞LDH的漏出率 ,分别为 4 8.2 %～ 89.3%和 6 2 .5 %～ 97.3% ,与对照组相比 ,差异有显著性 (P <0 .0 1 ) ,活细胞明显减少 ;应用流式细胞仪检测 ,在G1峰前出现亚二倍体峰 ,MA所致R2细胞凋亡呈一定的时间依赖性和剂量依赖性。DNA电泳图谱则表现出典型的梯状条带。 (2 )维拉帕米 (0 .5～ 5 0 μmol/L)减少MA导致的R2细胞LDH的漏出率 ,改善细胞形态 ,对MA诱导R2细胞凋亡有明显的抑制作用。结论 :维拉帕米对MA诱导的神经细胞凋亡具有保护作用 ,MA神经毒性作用可能与L型电压依赖性钙通道有关。
OBJECTIVE: To study the effect of verapamil on the damage of neurons induced by methamphetamine (MA) and to explore the mechanism of verapamil protection. Methods: MTT method and lactate dehydrogenase (LDH) method were used to observe the neurotoxic effect of MA. DNA electrophoresis and flow cytometry were used to observe the effect of MA on neuronal apoptosis. The protective effect of verapamil was also studied. Results: (1) MA (0.25 ~ 300 mmol / L) for 48 h and 60 h increased LDH leakage rate in rat cerebellar neuronal cell line R2, which were respectively 4.28% -89.3% and 6 2 .5% ~ 97.3%. Compared with the control group, the difference was significant (P <0.01), and the number of viable cells was significantly decreased. The results of flow cytometry showed sub-diploid peak before G1 peak, The apoptosis of R2 cells induced by MA showed a certain time-dependent and dose-dependent manner. DNA electrophoresis showed typical ladder-like bands. (2) Verapamil (0. 5 ~ 50 μmol / L) decreased the leakage rate of LDH induced by MA and improved cell morphology, and significantly inhibited the apoptosis of R2 cells induced by MA. CONCLUSION: Verapamil has a protective effect on MA-induced neuronal apoptosis. The neurotoxic effect of MA may be related to L-type voltage-dependent calcium channel.