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目的探讨尼莫地平能否诱导小脑颗粒神经元凋亡及可能机制。方法取Sprague-Dawley(SD)大鼠小脑颗粒神经元,体外培养7d,用含10μM尼莫地平培养基处理神经元24h,对照组为含1‰二甲亚砜(DMSO)培养基处理细胞。Hoechst33258染胞核检测凋亡率。Westernblot检测c-Fos和c-Jun蛋白的表达水平;腺病毒作载体过表达c-Fos和负显性c-Jun突变体阻断c-Jun功能,检测能否抑制尼莫地平诱导的神经元凋亡,免疫细胞化学方法检测腺病毒感染率。结果对照组神经元的凋亡率为(10±4)%,尼莫地平处理6、12、24h后神经元的凋亡率分别为(13±4)%、(28±5)%、(45±3)%。尼莫地平处理使c-Fos表达下调,但c-Jun表达水平上调。腺病毒Ad-c-Fos和Ad-c-JunDN(负显性c-Jun突变体)对神经元感染率为85%以上,过表达c-Fos或负显性c-Jun突变体使尼莫地平诱导的神经元凋亡率从(42±6)%减少到(28±6)%或(20±3)%。结论尼莫地平通过下调c-Fos和上调c-Jun表达诱导小脑颗粒神经元凋亡。
Objective To investigate whether nimodipine can induce apoptosis of cerebellar granule neurons and its possible mechanism. Methods Sprague-Dawley (SD) rat cerebellar granule neurons were cultured in vitro for 7 days. The neurons were treated with 10μM nimodipine for 24 hours, and the control group was treated with DMSO containing 1 ‰ dimethyl sulfoxide (DMSO). Hoechst33258 staining for apoptotic cells. Western blot was used to detect the expression of c-Fos and c-Jun protein. Adenovirus vector was used to overexpress c-Fos and negative dominant c-Jun mutant to block c-Jun function. Apoptosis and immunocytochemistry were used to detect the infection rate of adenovirus. Results The apoptosis rate of neurons in the control group was (10 ± 4)%. The apoptosis rates of neurons in the control group were (13 ± 4)%, (28 ± 5)%, 45 ± 3)%. Nimodipine treatment down-regulated c-Fos expression, but c-Jun expression was up-regulated. Ad-c-Fos adenovirus and Ad-c-JunDN (negative dominant c-Jun mutant) on the neuronal infection rate of more than 85%, overexpression of c-Fos or negative dominant c-Jun mutant so nimo The level of neuronal apoptosis induced by the horizon decreased from (42 ± 6)% to (28 ± 6)% or (20 ± 3)%. Conclusion Nimodipine induces apoptosis of cerebellar granule neurons by down-regulating c-Fos and up-regulating c-Jun expression.