肌肽对高糖诱导SH-SY5Y细胞凋亡的保护作用  

Protective effect of carnosine on high glucose-induced apoptosis in SH-SY5Y cells

作　　者：李宗泽[1] 杨清俊[1] 朱艳凌[1] 刘影[1] 杨菁[1] 李敏[1] 

机构地区：[1]辽宁医学院辽宁省心脑血管药物基础研究重点实验室,辽宁锦州121001

出　　处：《中国药理学与毒理学杂志》2013年第5期795-800,共6页Chinese Journal of Pharmacology and Toxicology

基　　金：辽宁省教育厅创新团队基金(2008T115)~~

摘　　要：目的探讨肌肽对高糖诱导的人神经母细胞瘤(SH-SY5Y)细胞凋亡的保护作用及其机制。方法 SH-SY5Y细胞用含葡萄糖50mmol·L-1的高糖培养液和肌肽0.6,3和15mmol·L-1的培养液培养48h,相差显微镜下观察细胞形态的变化;MTT比色法测定细胞存活率;Hoechst33258染色观察细胞核形态的变化;流式细胞仪检测细胞凋亡率;CDFH-DA探针法检测细胞中活性氧(ROS)的水平,酶联免疫吸附法检测细胞上清液中8-羟基脱氧鸟苷酸(8-OHdG)的含量。结果与正常组(低糖培养基)相比,高糖组细胞存活率明显降低(P<0.01);Hoechst33258染色发现细胞核固缩裂解,呈凋亡特征性改变;细胞凋亡率、细胞中ROS的水平、上清液中8-OHdG的分泌量明显升高(P<0.01)。与高糖组相比,肌肽0.6,3和15mmol·L-1组细胞存活率明显升高(P<0.01);Hoechst33258染色发现细胞核核固缩裂解减少,形态明显改善;细胞凋亡率分别降低至(20.65±1.13)%,(14.95±0.64)%和(9.95±0.61)%(P<0.01);细胞中ROS的水平分别降低至(182±16)%,(168±15)%和(140±10)%(P<0.05);上清液中8-OHdG的分泌量均明显降低,分别为2.780±0.154,2.136±0.100和(1.864±0.151)μg·L-1(P<0.01)。单纯肌肽组的细胞存活率、细胞核形态以及细胞凋亡率与正常组相比无显著差异,细胞中ROS水平和上清液中8-OHdG的分泌量均有所降低,分别为(88±9)%和(0.380±0.023)μg·L-1(P<0.05)。甘露醇组的各项指标与正常组相比无显著差异。结论肌肽对高糖诱导的SH-SY5Y细胞凋亡具有保护作用,其机制可能与肌肽的抗氧化作用有关。OBJECTIVE To investigate protective effects of carnosine on high glucose induced apoptosis and its potential mechanism. METHODS SH-SY5Y cells were cultured in medium （DMEM） which contained high glucose 50 mmol· L^-1 and carnosine 0.6, 3 and 15 mmol· L^-1 for 48 h. The changes in cell morphology were observed under an inverted phase contrast microscope, the cell viability was detected by MTT assay, and changes in nucleus were assayed by Hoechst33258 staining. The apoptosis rate was measured by flow cytometry using Annexin V and propidium iodide （PI）. The reactive oxygen species （ROS） level in cells was detected by fluorescent probe CDFH-DA, while the content of 8-OHdG in culture medium was determined by enzyme-linked immunosorbent assay. RESULTS Compared with normal control group, the cell viability in high glucose group decreased（ P〈 0.01 ）, the karyopyknosis, split and apoptosis were found by Hoechst33258 dyeing; while the apoptosis rate and content of 8-OHdG were increased. Compared with high glucose group, the cell viability of carnosine 0.6, 3 and 15 mmol· L^-1 groups obviously increased（ P 〈0.01 ） ; while the karyopyknosis and split decreased; the form of cells significantly improved; the apoptosis rate was respectivedly reduced to （20.65±1.13）%, （14.95 ±0.64）% and （9.95±0.61）%（P〈0.01 ）; the level of ROS in the cells was reduced to（182±16）% ,（168 ±15）% and （140 ±10）% （P〈0.05） ; and the content of 8-OHdG in culture medium was reduced to 2.780±0.154,2.136 ±0.100 and （1.86 ±0.15） pg.L-1 （P 〈0.01 ）. Compared with normal control group, the cell viability, change in nucleus and apoptosis rate did not significantly change in carnosine group; ROS levels and contents of 8-OHdG were respectively reduced to （88± 9） % and （0. 380±0. 023） ug· L^-1 （ p 〈 0.05）. There was no significant difference between the normal group and the mannitol group. CONCLUSION Carnosine has protective effect on high glucose induced apoptosis. T

关 键 词：高糖 SH—SY5Y细胞 细胞凋亡 肌肽 活性氧 8-羟基脱氧鸟苷酸 

分 类 号：R965[医药卫生—药理学]