摘要
目的通过检测L929细胞中的衰老及自噬水平,探讨欧亚旋覆花总黄酮(IBFTF)对L929细胞衰老和自噬的影响,以更好地开发IBFTF的药用价值。方法分别使用0、5、10、20、40 g/L D-半乳糖作用于L929细胞,探究D-半乳糖诱导L929细胞衰老的最适浓度,分别使用浓度为25、50、100 mg/L IBFTF作用于衰老L929细胞探究IBFTF对L929细胞衰老和自噬的影响,通过MTT检测不同浓度D-半乳糖对L929细胞活力的影响,β-半乳糖苷酶染色检测不同浓度的D-半乳糖对L929细胞衰老进程的影响,荧光显微镜检测IBFTF对衰老L929细胞内活性氧生成的影响,通过吖啶橙染色和免疫荧光探究IBFTF对衰老L929细胞中溶酶体数量和LC3蛋白的表达,用Western-Blot检测细胞自噬相关蛋白水平。结果 IBFTF呈剂量依赖性延缓衰老L929细胞的衰老进程,在延缓衰老过程中L929细胞自噬水平也呈现剂量依赖性增高。结论 IBFTF延缓衰老L929细胞衰老的过程可能与自噬水平的上调有关。
Objective To investigate the effect of Inula britannica flower total flavonoids(IBFTF) on autophagy of aging L929 cells by detecting the levels of autophagy in the process of senescence of L929 cells for better development of the medicinal value of IBFTF. Methods Cell senescence models were established by using D-galactose(D-gal) solutions(0, 5, 10, 20 and 40 g/L) on L929 cells to find out the most appropriate concentration of D-gal for inducing senescence in L929 cells. IBFTF solutions(25, 50, 100 mg/L) were applied to aging L929 cells to investigate the effect of IBFTF on L929 cell senescence and autophagy. The effect of D-gal on the viability of L929 cells was detected with MTT. β-galactosidase staining was used to detect the effect of D-gal at different concentrations on the senescence process of L929 cells. Fluorescence microscopy was used to detect the effect of IBFTF on ROS production in senescent L929 cells. The effects of IBFTF on the number of lysosomes and the expression of LC3 protein in senescent L929 cells were examined with acridine orange staining and immunofluorescence. The levels of autophagy-related proteins were detected with Western-Blot. Results IBFTF delayed the senescence process of aging L929 cells in a dose-dependent manner, and the autophagy level of L929 cells also increased in a dose-dependent manner during the aging delay process. Conclusion IBFTF delayed the senescence of D-gal-induced senescent cells, which may be related to the elevated levels of autophagy.
作者
郭宗雨
刘露
况杰
周茂强
果磊
Guo Zongyu;Liu Lu;Kuang Jie;Zhou Maoqiang;Guo Lei(Department of Burn and Plastic Surgery,The First Affiliated Hospital of Chongqing Medical University,Chongqing 400016,China)
出处
《北京中医药大学学报》
CAS
CSCD
北大核心
2019年第12期1024-1029,共6页
Journal of Beijing University of Traditional Chinese Medicine
基金
重庆市科卫联合中医药科研项目(No.2019ZY023405).
关键词
欧亚旋覆花总黄酮
L929细胞
自噬
衰老
Inula britannica flower total flavonoids
L929 cells
autophagy
aging
作者简介
郭宗雨,男,在读硕士生;通信作者:果磊,女,博士,副教授,主任医师,硕士生导师,主要研究方向:中西医结合抗衰老研究,E-mail:841602937@qq.com
