摘要
目的:观察黄芪甲苷对长波紫外线(ultraviolet A,UVA)辐射导致的人成纤维细胞衰老的抑制作用,以及对基质金属蛋白酶-1(matrix metalloproteinase-1,MMP-1)和金属蛋白酶组织抑制因子-1(tissue inhibitor of metalloproteinase-1,TIMP-1)等老化相关基因表达的影响。方法:分离培养人原代成纤维细胞,将亚融合状态的培养细胞分为空白对照组、黄芪甲苷组、UVA组和UVA+黄芪甲苷组,以10J/cm2UVA进行照射,并加入20μg/mL黄芪甲苷干预处理。采用组织化学染色法检测衰老相关β半乳糖苷酶表达,以酶联免疫吸附法检测上清液中转化生长因子-β1(transforming growth factor-β1,TGF-β1)的含量,实时聚合酶链反应法检测MMP-1和TIMP-1的mRNA表达水平变化。结果:空白对照组及黄芪甲苷组β半乳糖苷酶阳性细胞均较低,UVA组β半乳糖苷酶阳性细胞数则显著升高,加入黄芪甲苷处理可使β半乳糖苷酶阳性细胞比率明显降低(P<0.05)。黄芪甲苷处理可以促进成纤维细胞分泌TGF-β1;UVA照射成纤维细胞后TGF-β1分泌量明显降低,UVA照射前加入黄芪甲苷可增加TGF-β1分泌量(P<0.05)。此外,UVA能够诱导MMP-1和TIMP-1的mRNA表达水平升高,而黄芪甲苷可在一定程度上抑制MMP-1mRNA表达,并诱导TIMP-1mRNA表达(P<0.05)。结论:黄芪甲苷可有效延缓人成纤维细胞光老化进程,其机制可能与促进TGF分泌和抑制胶原降解相关。
Objective：In this study,we aim to investigate the protective effects of astragaloside on ultraviolet A（UVA）-induced photoaging in human fibroblasts and its possible mechanisms.Methods：Subconfluent fibroblasts were cultured and divided into normal control group,astragaloside group,UVA irradiation group and UVA plus astragaloside group.The cells were shammed or irradiated with 10 J/cm2 of UVA irradiation and treated with 20 μg/mL astragaloside.The aging condition was determined by histochemical staining of senescence-associated β-galactosidase（SA-β-gal）.Concentration of transforming growth factor-β1（TGF-β1）in the supernatant was determined by enzyme-linked immunosorbent assay,and mRNA levels of matrix metalloproteinase-1（MMP-1）and tissue inhibitor of metalloproteinase-1（TIMP-1）were measured by real-time polymerase chain reaction Results：UVA irradiation raised the proportion of SA-β-gal-positive cells in comparison with the normal control group（P0.05）.Astragaloside treatment was shown to decrease the level of SA-β-gal compared with the UVA group.With UVA irradiation,the concentration of TGF-β1 in the supernatant decreased,and astragaloside treatment recovered the content of TGF-β1 compared with the UVA irradiation alone（P0.05）.UVA irradiation also up-regulated the mRNA levels of MMP-1 and TIMP-1（P0.05）.Astragaloside decreased the mRNA level of MMP-1 compared with the UVA irradiation alone,while the TIMP-1 expression increased（P0.05）.Conclusion：Astragaloside can protect the skin from UVA irradiation.The mechanism involved may be related with TGF secretion and decrease of collagen degradation.
出处
《中西医结合学报》
CAS
2011年第3期328-332,共5页
Journal of Chinese Integrative Medicine
