摘要
目的 研究D-半乳糖处理后引起人正常二倍体细胞衰老的机制.方法 MTT法检测细胞存活率,衰老相关的β-半乳糖苷酶染色鉴定衰老细胞,流式细胞术检测细胞内活性氧自由基水平,免疫印迹法检测衰老相关蛋白的表达.结果 D-半乳糖处理人胚胎肺细胞和肝细胞,均能抑制细胞的增殖.被处理的细胞衰老相关的β-半乳糖苷酶染为蓝色,细胞内活性氧自由基水平明显升高,衰老信号通路相关蛋白p53、p21、caviolin-1的表达均升高.结论 D-半乳糖引起人正常二倍体细胞出现典型的细胞衰老表型,可以作为衰老的可靠模型.
Objective To investigate the mechanism of cellular senescence induced by D-galactose in normal human diploid cells. Methods Cell viabilities were detected using MTT. Senescence-associated β-galactosidase activity assay was performed to show the senescent cells. The levels of reactive oxygen species were detected by flow cytometry. The levels of aging-related proteins were detected by immunoblotting assay. Results The growth-inhibitory actions of D-galactose were detected in the human embryonic lung fibroblast cells and liver cells. The blue cells were observed with galactosidase staining in the D-galactose -treated cells. The increased level of intracellular reactive oxygen species had been detected in the cells after exposure to D-galactose. The protein levels of p53, p21 and caviolin-1 were elevated with identification of immunblotting. Conclusions D-galactose can induce the typical features of cellular senescence in human diploid cells and it is a reliable model of aging.
出处
《老年医学与保健》
CAS
2014年第6期372-375,共4页
Geriatrics & Health Care
基金
国家自然科学基金（31471150）
