尼莫地平含药脑脊液对体外PC12细胞损伤的保护作用

Neuroprotective Effect of Cerebrospinal Fluid of Nimodipine on Various PC12 Cell Injuries in Vitro

关键词:  脑脊液药理  尼莫地平  条件规范化  神经保护

Key words:  cerebrospinal fluid pharmacology  nimodipine  method standardization  neuroprotective effect

摘要:

目的 观察尼莫地平含药脑脊液对多种PC12细胞损伤模型的保护作用，对脑脊液药理实验条件的规范化及其应用进行探讨。方法 采用噻唑蓝（MTT）法，观察不同取样时间、不同添加量、不同灭活方法处理的尼莫地平大鼠含药脑脊液对氯化钾损伤的PC12细胞存活力的影响，确定尼莫地平大鼠含药脑脊液的制备条件。并进一步考察该条件制备的尼莫地平大鼠含药脑脊液对过氧化氢、连二亚硫酸钠及谷氨酸钠等因素引起的PC12细胞损伤的保护作用。结果 尼莫地平含药脑脊液对氯化钾致PC12细胞损伤的保护率随添加量的增加而提高；相同添加量多次给药（2次·d-1×3.5 d）含药脑脊液的保护率均高于单次给药的含药脑脊液；热灭活、乙醇灭活及丙酮灭活含药脑脊液对PC12细胞均有显著保护作用，其保护率与未灭活含药脑脊液无显著性差异；根据确定条件制备的尼莫地平大鼠含药脑脊液对过氧化氢、连二亚硫酸钠及谷氨酸钠等因素损伤的PC12细胞均具有显著的保护作用。结论 尼莫地平含药脑脊液的制备以连续多次给药方案，无需灭活处理，体外添加终体积的10%为宜；对氯化钾等多种原因引起的PC12细胞损伤均具有明显保护作用。

Abstract:

OBJECTIVE To explore the standard conditions and application of the cerebrospinal fluid （CSF）pharmacological method, by investigating the neuroprotective effect of CSF with nimodipine on PC12 cells injured by various factors. METHODS The effect of rat CSF with nimodipine on injured PC12 cells induced by potassium chloride (KCl) was observed by 3-(4,5-dimethyl- 2-thiazol-2, 5-diphenyl-2H-tetrazolium bromide (MTT assay under various experimental conditions (dosage regiment, addition level and treatment process, then its neuroprotective effect under certain conditions on PC12 cells injured by hydrogen peroxide (H2O2, sodium dithionite (Na2S2O4 and L-glutamate sodium were determined as well. RESULTS The viability of PC12 cells injured by KCl increased with the addition level of CSF with nimodipine, and at the same addition level, CSF of rat administrating nimodipine repetitiously (twice·d-1×3.5d) exerted higher protective efficacy on PC12 cells than that of single administration.The raw CSF with nimodipine and the CSF with nimodipine treated by heat (56 ℃), ethanol or acetone had notable protective effects on injured PC12 cells (P<0.01, and there was no obvious statistical difference among them. The similar distinct protective effects of CSF with nimodipine under certain conditions on PC12 cells injured by H2O2, Na2S2O4 and L-glutamate sodium were also observed. CONCLUSION It is proposed that the CSF with nimodipine having significant protective effects on PC12 cells injuries caused by multi-factors such as KCl, Na2S2O4 and H2O2 etc may be prepared by 10 rat CSF without treating after administrating drugs repetitiously.