仙草提取物对小鼠脾淋巴细胞DNA氧化损伤保护作用的研究

Protection of Hsian-tsao(Mesona procumbens Hemsl)on H_(2)O_(2)-induced DNA damage of primarily cultured mice spleen lymphocytes

目的:观察仙草提取物对原代培养脾淋巴细胞H2O2所致DNA氧化损伤的保护作用。方法:原代培养24 h的脾淋巴细胞悬液,随机分为6组,其中4组细胞用不同浓度的仙草提取物溶液预先处理60 m in,再加入50μm o l/L的H2O2,H2O2染毒组直接加入相同浓度的H2O2,空白对照组加入等量的PBS溶液,6组细胞共同在4℃下染毒20 m in,收获细胞同时进行单细胞凝胶电泳,最后用激光共聚焦显微镜拍摄图像,计算DNA迁移的细胞率和总彗星长度。结果:H2O2可致原代培养脾淋巴细胞DNA的严重损伤,而仙草提取物能不同程度地降低H2O2诱导产生的DNA损伤,在10、50、100μg/m l的浓度下,彗星细胞出现率从阳性对照组的100%分别降低为88%、60%和36%(P分别<0.05、0.01、0.01),总彗星长度也从对照组的(49.56±6.94)μm,逐渐降低为(41.14±5.64)μm、(38.89±9.81)μm、(28.62±4.66)μm(P分别<0.05、0.01、0.01)。结论:仙草提取物具有显著的抗氧化功能,能在一定浓度范围内保护细胞免受氧自由基对DNA的氧化损伤。

Objective： To investigate the protection of Hsian-tsao （Mesona procurnbens Hems/） on H2O2-induced DNA damage of primarily cultured spleen lymphocytes. Methods： Spleen lymphocytes were primarily cultured for 24 h and then divided into 6 groups randomly：4 groups were treated with different concentrations of Hsian-tsao extracts for 60 rain,then with 50μmol/L H2O2; 1 group was treated with 50μmol/L H2O2 only and the last group with PBS. Twenty rain after H2O2 or PBS treatment SCGE and laser confocal microscopy were performed ,then the comet length and percentage of cells with migrated DNA were measured. Results： H2O2 caused severe DNA damage in primarily cultured spleen lymphocytes and Hsian-tsao extracts reduced the DNA damage at a concentration of 10.50 and 100μg/ml. The quantity of comet cells was reduced from 100% to 88%,60%,36% （P〈0. 05,0. 01,0. 01） and the comet length was reduced from （49.56±6.94）μm to （41. 14±5.64）μm, （38.89±9.81）μm, （28.62±4.66）μm（P〈0.05,0. 01,0. 01）, respectively. Conclusion： Hsian-tsao has dramatic ability of antioxidant. It can protect cells from the damage of ROS-induced DNA injury in the certain range.