Vam3对小鼠体外胸腺细胞的辐射防护效应

Radiation protection effects of Vam3 on mouse thymocyte in vitro

探讨白藜芦醇二聚体(Amurensis H,Vam3)对辐射致小鼠胸腺细胞损伤的保护作用。体外无菌分离小鼠胸腺细胞,实验分为6组:对照组(Control)、白藜芦醇组(Resveratrol,Res)、Vam3组、照射组(Irradiation,IR)、照射+Res组和照射+Vam3组。照射+Res组和照射+Vam3组于照射前30 min给予Vam3和Res孵育,对照组、Res组、Vam3组以及照射组加等量RPMI-1640培养基或对应药物处理,除对照组、Res组和Vam3组外,其余3组均给予4 Gy ^(137)Cs γ-射线单次照射。照射后分别用生物发光法检测小鼠胸腺细胞活力,活性氧探针(DCFH-DA)检测细胞内活性氧(Reactive oxygen species,ROS)水平,荧光抗体标记检测γ-H2AX表达水平以及FITC-Annexin V和PI双染标记法检测细胞早期凋亡率。结果显示,与对照组比较,照射组细胞活力显著下降,细胞内ROS和γ-H2AX水平明显升高,同时细胞早期凋亡数目增加;而照射+Vam3组与照射+Res组比较显示,Vam3在提升细胞活力,降低细胞内ROS和γ-H2AX水平以及抑制细胞凋亡方面的作用比Res更加明显。研究结果表明,Vam3对辐射引起的小鼠胸腺细胞急性损伤有一定保护作用,且保护作用优于白藜芦醇。

The aim is to observe the protective effect of Vam3 on irradiation-induced injury of thymocytes isolated sterilely from mice. The thymocytes were divided into 6 groups： control group, resveratrol group, Vam3group, irradiation group, irradiation with resveratrol group and irradiation with Vam3 group. Except the control group, resveratrol group and Vain3 group, the rest groups were exposed to radiation at a single dose of 4 Gy 7-rays from 137Cs after 30 min treatment by vehicle and tested drug, respectively. The thymocytes＇ viability was measured by bioluminescence; the level of thymocytes＇ reactive oxygen species （ROS） was measured by DCFH-DA; the level of thymocytes＇ 7-H2AX was measured by fluorescent antibody labeling; the early apoptosis was detected by F1TC-Annexin V and PI labelling using flow cytometry （FCM） after radiation. Compared with control group, the thymocytes＇ cell viability decreased obviously after irradiation, and the level of cell ROS, Y-H2AX and early apoptosis cell increased. By comparing the resveratrol group and Vam3 group with the irradiation group, we can find that Res and Vain3 could reduce the decrease of thymocytes＇ cell viability and inhibit the increase of ROS,Y-H2AX and early apoptosis cell induced by irradiation. The comparison between irradiation with resveratrol group and irradiation with Vam3 group showed that the effects of Vain3 in increasing cell viability, decreasing the level of ROS and 7-H2AX and lowering the early apoptosis rate were more obvious than resveratrol. It can be concluded that Vam3 performs better than resveratrol in the protection of mice thymocytes against the acute radiation injury.