摘要
目的探讨淫羊藿总黄酮 (EF)延缓人胚肺二倍体成纤维细胞衰老以及保护端粒缩短的作用机制。方法采用含EF的血清对人二倍体成纤维细胞 2BS细胞株进行处理 ,观察 2BS细胞寿命 ;采用荧光实时定量PCR法检测 p16基因mRNA的表达 ;ELISA法检测细胞视网膜母细胞瘤 (retinoblastoma ,Rb)蛋白和磷酸化Rb蛋白的含量 ;TRAP Hyb单管一步法检测细胞端粒酶活性 ;端粒限制性片段 (TRF)Southernblot法检测 2BS细胞端粒长度变化。结果淫羊藿总黄酮能够延长 2BS细胞的传代寿命 ;下调 2BS细胞p16基因mRNA的表达 ;增加磷酸化Rb蛋白的含量 ;延缓衰老细胞端粒长度的缩短 ,而不是激活细胞端粒酶活性。结论 2BS细胞衰老时 ,p16基因mRNA的表达增加 ,磷酸化Rb蛋白含量减少 ,细胞端粒长度缩短 ;淫羊藿总黄酮可能通过抑制p16基因表达 ,促进磷酸化Rb蛋白的产生 ,从而延缓衰老细胞端粒长度的缩短 ,具有延缓细胞衰老的作用 ,而不是通过激活细胞端粒酶的活性。
ObjectiveTo investigate the mechanism of senes cence delay of human d iploid fibroblast (2BS) and protecting telomere length by epimedium flavonoids ( EF). MethodsThe drug sera of EF were used to treat the 2BS. The population dou blings of 2BS cells were observed, the mRNA expression of p16 gene were determin ed by fluorescence real-time quantitative RT-PCR, the telomerase activation of 2 BS cells were determined by TRAP-Hyb, the total retinoblastoma (Rb) and phospho r ated Rb protein content were detected by ELISA, the telomere length of 2BS cells were determined by telomere restriction fragment (TRF) Southern blot assay. ResultsEF could significantly extend the population doubli ngs of 2BS cells, the e xpression of p16 mRNA was decreased and the content of phosphorated Rb protein w ere increased by EF. The telomere lengthening of 2BS cells were improved by EF, but the telomerase was not activated. ConclusionIn se nescence human fibroblast s 2BS cells, p16 gene mRNA expression increased, content of phosphorated Rb prot ein decreased and the telomere length of 2BS shortened, EF might delay the aging of cells through inhibit ing the p16 gene expression, promoting the production of phosphorated Rb protein and to protect the length of telomere, but not activat ing the telomerase.
出处
《中国中西医结合杂志》
CAS
CSCD
北大核心
2004年第12期1094-1097,共4页
Chinese Journal of Integrated Traditional and Western Medicine
基金
国家自然科学基金资助项目 (No .30 2 71 666)
