生物发光法测定PC12细胞微量ATP含量

目的　研究生物发光法测定PC12细胞内腺苷三磷酸 (ATP)含量的可行性及短暂缺氧血清剥夺再灌注后ATP含量的动态变化。方法　将PC12细胞随机分为正常对照组、缺氧组 ,测定短暂缺氧血清剥夺再灌注后不同时间点的ATP含量和细胞活性 ,对缺氧组ATP含量与细胞活性进行相关性分析。结果　与正常对照组相比 ,缺氧血清剥夺 15min再灌注1h后的ATP含量、细胞活性显著降低 ,差异具有显著性意义 (P <0 .0 5 ) ;缺氧血清剥夺 15min再灌注 6h后基本恢复至正常水平。缺氧组ATP含量与细胞活性呈显著正相关 (r=0 .90 4 ,P <0 .0 5 )。结论　生物发光法测定PC12细胞内ATP含量有较高的准确性。短暂缺氧血清剥夺再灌注后 ,存在低能量状态 ,且可完全恢复。

Detection of ATP content in PC12 cells by bioluminescence method

Objective To study the feasibility of detecting ATP content in PC12 cells by bioluminescence method and the dynamic changes of ATP content after transient anoxia-serum deprivation/reperfusion.Methods The PC12 cells were randomly divided into control group and anoxia group. The content of ATP and cell activity were determined at different time points after transient anoxia-serium deprivation/reperfusion. The correlation between ATP content and cell activity in anoxia groups was analyzed.Results The content of ATP and cell activity at 15 min after anoxia-serum deprivation/1 h after reperfusion in the anoxia group were significantly lower than in the control group (P< 0.05 ) and they returned to the normal level at 15 min after anoxia-serum deprivation/6 h after reperfusion. There was a positive correlation between ATP content and cell activity (r= 0.904 , P< 0.05 ).Conclusion Bioluminescence method can detect ATP content in PC12 cells accurately. There existed a lower energy status after transient anoxia-serum deprivation and reperfusion and the lower energy status can resume the normal level.