Objective To study the influences of NADH on the intracellular reactive oxygen species(ROS), free Ca2+and pH value in X rays-treated L02 cells. Methods L02 cells was cultured in vitro and divided into three groups. L02 cells were cultured in RPMI 1640, exposed to 5.0 Gy X-ray irradiation and continued to culture in the presence or absence of NADH with the concentration of 400 μg·mL-1 for 24 h, which was respectively named irradiation group and treatment group. The control group, without undergoing X-ray irradiation, was cultured in the absence of NADH. The apoptotic rate was determined by PI/Annexin V staining method. The level of intracellular reactive oxygen species, intracellular free Ca2+value and pH value, were detected by confocal microscopy. Results The apoptotic rates of treatment group and irradiation group were respectively (8.25±1.64)%and (28.16±2.46)%. The apoptotic rate was significantly lower in treatment group than in irradiation group (P<0.05). The mean value of intracellular ROS, free Ca2+and pH in irradiation group were signifi-cantly increased, as compared with those of the control group(P<0.05). But the value of intracellular ROS, free Ca2+and pH were obviously lower in treatment group than in irradiation group (P<0.05). Conclusion NADH could obviously inhibit the apoptosis of L02 liver cell line induced by X-ray irradiation, which might be associated with the down-regulation of intrac-ellular ROS, Ca2+and pH value.%目的：探讨烟酰胺腺嘌呤二核苷酸（NADH）对正常肝细胞L02辐射后细胞内活性氧簇（ROS）、Ca2+和pH的影响。方法采用体外培养的正常肝细胞L02为研究对象，并将其分为处理组、辐射组和对照组。处理组和辐射组细胞均给予5.0 Gy X射线辐射，随后分别在加/不加NADH（400μg·mL-1）的培养液中培养24 h，对照组仅给予RPMI 1640完全培养基培养24 h。采用PI/Annexin V双标染色法检测细胞凋亡率，共聚焦显微镜检测细胞内ROS、Ca2+和pH值。结果处理组和辐射组的细胞凋亡率分别为（8.25±1.64）％和（28.16±2.46）％，辐射组显著高于处理组，差异具有统计学意义（P<0.05）。与对照组相比，辐射组细胞内ROS、Ca2+和pH值明显升高（P<0.05），而处理组较辐射组显著下降，差异具有统计学意义（P<0.05）。结论 NADH能够抑制辐射诱导L02肝细胞凋亡，其作用机制可能与下调细胞内ROS、Ca2+和pH有关。
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