利用X射线辐照经羧甲基-β-1,3-葡聚糖预处理的人B淋巴母细胞(Human B lymphoblasts,HMy2.CIR),探究羧甲基-β-1,3-葡聚糖对辐射损伤的防护作用。采用CCK-8法检测细胞存活率,并筛选出最佳给药浓度和孵育时间(0.1μg/mL,72 h);流式细胞仪检测羧甲基-β-1,3-葡聚糖对辐照后HMy2.CIR细胞凋亡的影响;微核实验检测羧甲基-β-1,3-葡聚糖对辐照后HMy2.CIR细胞微核形成的影响;彗星实验检测对DNA损伤程度和修复的影响。结果表明,羧甲基-β-1,3-葡聚糖对HMy2.CIR无细胞毒性并具有增殖促进作用;羧甲基-β-1,3-葡聚糖能够抑制辐射造成的凋亡率和微核率的增加,降低DNA损伤程度,加快损伤DNA的修复。羧甲基-β-1,3-葡聚糖对HMy2.CIR细胞辐射损伤有防护作用,其机制可能与抑制辐射诱导的细胞凋亡和DNA损伤有关。
The aim is to investigate radiation protection effect induced by carboxymethyl-β-1,3-glucan on Human B lymphoblasts(HMy2.CIR). The survival rate of carboxymethyl-β-1,3-glucan on HMy2.CIR was tested by CCK-8 assay at concentrations of 0, 0.01, 0.1, 1,100, 200 and 400 μg/mL, respectively. The flow cytometry was used to measure the apoptosis rate. Micronucleus rate was assessed by the in vitro micronucleus test. DNA strand breaks were assessed by the comet assay. The results showed that treatment with carboxymethyl-β-1,3-glucan was not cytotoxic. Pre-treatment with carboxymethyl-β-1,3-glucan at concentration of 0.1 μg/mL for 72 h protected HMy2.CIR cells against radiation, as indicated by increased surviving fraction, reduced apoptosis, reduced micronucleus rate and fewer DNA strand breaks. The carboxymethyl-β-1,3-glucan has radiation protection effect onHMy2.CIR cells, which may be related to inhibition of apoptosis and DNA damage after radiation.