马来酸桂哌齐特对不同条件下体外培养的人脑胶质瘤U251细胞腺苷激酶的影响

Effects of Cinepazide Maleate on the Expression of Adenosine Kinase in Human Glioma U251 Cells Cultured in Vitro Under Different Conditions

目的观察马来酸桂哌齐特(CM)对不同条件下体外培养的人脑胶质瘤U251细胞腺苷激酶(ADK)表达水平的影响,探讨CM是否可通过影响细胞内ADK表达水平而实现其神经保护作用。方法 2013年2月—2014年6月,将人脑胶质瘤U251细胞铺到6孔板,1孔为1组,分别为胎牛血清(FBS)+0.9%氯化钠溶液对照组(C组)、FBS+CM 0.5 mg/ml组(C+0.5组)、FBS+CM 1.5 mg/ml组(C+1.5组)、血清剥夺(SD)+0.9%氯化钠溶液对照组(SD组)、SD+CM 0.5 mg/ml组(SD+0.5组)、SD+CM 1.5 mg/ml组(SD+1.5组),进行正常体外细胞培养或SD体外缺血模式培养。提取细胞蛋白、RNA,采用Western blotting法检测人脑胶质瘤U251细胞ADK表达水平,RT-PCR法检测人脑胶质瘤U251细胞ADK mRNA表达水平。结果 6组ADK表达水平比较,差异有统计学意义(P<0.05)。SD+0.5组ADK表达水平低于C组(P<0.05)。6组ADK mRNA表达水平比较,差异有统计学意义(P<0.05)。SD+0.5组ADK mRNA表达水平低于C组、SD+1.5组(P<0.05)。结论 SD体外缺血模式培养下,CM在一定浓度范围内可能通过增强人脑胶质瘤U251细胞的ADK表达抑制作用来实现其神经保护作用。

Objective To observe the effects of cinepazide maleate（ CM） on adenosine kinase（ ADK） in human glioma U251 cells cultured in vitro under different conditions. Methods During the period from February 2013 to June 2014,human glioma U251 cells were spread into 6 holes of a 6 welll plate,with 6 holes being 6 different groups： fetal bovine serum（ FBS） ＋ 0. 9% sodium chloride solution control group（ C group）,FBS ＋ CM 0. 5 mg/ml group（ C ＋ 0. 5 group）,FBS ＋ CM1. 5 mg / ml group（ C ＋ 1. 5 group）,SD ＋ 0. 9% sodium chloride solution control group（ SD group）,SD ＋ CM 0. 5 mg / ml group（ SD ＋ 0. 5 group） and SD ＋ CM 1. 5 mg/ml（ SD ＋ 1. 5 group）. Normal in vitro cell culture or SD in vitro ischemia model culture were conducted. Matricellular protein and RNA were extracted. Western blotting method was employed to detect the expression level of ADK in human glioma U251 cells,and RT-PCR method was adopted to determine the mRNA expression level of ADK in human glioma U251 cells. Results The 6 groups were significantly different in the expression level of ADK（ P〈0. 05）. SD＋ 0. 5 group was lower than C group in the expression level of ADK（ P〈0. 05）. The 6 groups were significantly different in the mRNA expression level of ADK（ P〈0. 05）. SD ＋ 0. 5 group was lower（ P〈0. 05） than C group and SD ＋ 1. 5 group in the mRNA expression level of ADK. Conclusion In SD in vitro ischemia model culture,CM could exert its neuroprotective effect in a certain concentration range by increasing inhibition of the expression of ADK in human glioma U251 cells.