水溶性低分子量壳聚糖对辐射诱导正常肝细胞株BRL损伤的影响

摘要

Objective To investigate the effects of water-soluble low molecular weight chitosan (WSC) on the irradiated damage of normal liver cell line BRL. Methods BRL cells were cultured in vitro and divided into six groups of A(4 Gy 60Co γ-rays) ,B1(4 Gy 80Co γ-rays+WSC 1. 56 μg/ml) ,B2 (4 Gy 60Co γ-rays+WSC 12. 50 μg/ml) ,B3(4 Gy 60Co γ-rays+WSC 25. 00 μg/ml) ,B4(4 Gy 60Co γ-rays+WSC 50.00 μg/ml) and C(normal controls). At 24 h and 48 h after irradiation, the cell apoptosis and fluorescence intensity of intracellular Ca24 were detected by Annexin V-FITC/PI double staining and Fluo-3AM, respectively. The survival fraction of BRL cells was analyzed by colony formation assay. The protection factor( PF) of WSC on BRL cells was calculated and evaluated with multi-target one shot model. Results Compared with group C, the rate of cell apoptosis and fluorescence intensity of intracellular Ca2+ were increased in group A (P<0. 05), which were significantly reversed in groups of B1,B2, B3 and B4(F<0. 05). There was an excellent correlation between cell apoptosis rate and fluorescence intensity of intracellular Ca2+ (y=l. 03x-7. 47 and R2 = 0. 9994). Colony formation assay showed that the PF was more than 1 after WSC treatment. Conclusion WSC can inhibit the apoptosis of BRL cells induced by irradiation, which may be related to the effect of inhibiting intracellular calcium overload.%目的 研究水溶性低分子量壳聚糖(WSC)对辐射照射后正常肝细胞株BRL损伤的影响.方法 体外培养的BRL细胞分为4 Gy照射组(A组)、4 Gy照射+WSC 1.56、12.50、25.00、50.00 μg/ml组(分别为B1、B2、B3、B4组)和对照组(C组).照射后24、48 h,采用Annexin V-FITC/PI双染法和Flou-3AM法分别检测BRL细胞凋亡和胞内钙离子荧光强度.克隆形成实验分析BRL细胞生存率的变化,以多靶单击数学模型拟合方程计算WSC对细胞的保护系数(PF)并评价其辐射防护效果.结果 与C组相比,A组细胞凋亡率、胞内钙离子荧光强度增加(P＜0.05),而WSC能明显逆转上述指标的变化(P＜0.05).细胞凋亡率和胞内钙离子浓度呈良好的线性关系(y=1.03x-7.47,R2=0.9994).克隆形成实验结果表明,WSC处理后的各组PF值均大于1.结论 WSC能抑制电离辐射诱导的细胞凋亡,可能与WSC抑制细胞内钙超载有关.