富H_2溶液对人肾小管上皮细胞辐射损伤的防护作用

Protective effects of hydrogen-rich saline againstγ-ray irradiation-induced injury in kidney epithelial HK-2 cells

目的探讨富H2溶液对人肾小管上皮细胞(HK-2细胞)辐射损伤的防护效应及机制。方法将HK-2细胞分为4组:对照组、H2组、照射组和照射加H2组。对照组和照射组细胞加入5mL无H2的PBS溶液,H2组和照射加H2组细胞则加入等量饱和富H2溶液,孵育10min后将照射组和照射加H2组利用60 Coγ射线以6Gy剂量照射325s,随即用比色法测定细胞丙二醛(MDA)含量变化;37℃条件下与羟苯基荧光素(HPF)荧光探针孵育15min后测定细胞内羟自由基(·OH)水平;照射后24h用流式细胞仪检测细胞凋亡情况;照射后每天计数各组细胞克隆形成以反映各组细胞存活率,观察10d。结果富H2溶液可以提高HK-2细胞经6Gy照射后第10天的细胞存活率(P<0.05),并且能明显减少照射后24h细胞凋亡水平(P<0.01);富H2溶液还可以降低受照细胞的MDA水平(P<0.05)以及·OH水平(P<0.05)。结论富H2溶液对60 Coγ射线导致的HK-2细胞辐射损伤有较好的防护作用。

Objective To investigate the protective effects and mechanism of hydrogen-rich saline againstγirradiationinduced injury in kidney epithelial HK-2cells.Methods HK-2cells were divided into 4groups:the control group,the hydrogen-treated group,the irradiated group,and the irradiation plus hydrogen-treated group.The hydrogen-treated group and the irradiation plus hydrogen-treated group were pretreated with 5mL hydrogen-rich solution while the control group and the irradiated group with 5mL non-hydrogen PBS for 10 min,then the irradiated group and the irradiation plus hydrogen-treated group were simultaneously treated withγ-irradiation(6Gy)for 325 s.The malondialdehyde(MDA)changes were determined by clorimetric method,and the hydroxyl radicals(·OH)were examined after being incubated with HPF for 15 min at 37℃.Flow cytometry was used to observe the apoptosis rates of HK-2cells 24 hafter irradiation,and the cell survival was examined by clonogenic assay every 24 huntil the 10 th day.Results Hydrogen-rich saline significantly improved the survival of HK-2cells at the 10 th day after 6Gy irradiation(P<0.05),and significantly decreased the apoptosis rates of HK-2cells(P<0.01).It was also found that hydrogen-rich saline significantly down-regulated MDA level(P<0.05)and ·OH level in irradiated HK-2cells(P<0.05).Conclusion Hydrogen-rich saline has a protective effect againstγ-irradiation-induced injury in HK-2cells.