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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: To observe the inhibitory effect of astragaloside on the aging of human fibroblasts induced by ultraviolet A (UVA) radiation and the effects of matrix metalloproteinase-1 (MMP-1) and metalloproteinase tissue inhibitor -1 (tissue inhibitor of metalloproteinase-1, TIMP-1) and other aging-related gene expression. Methods: Human primary fibroblasts were isolated and cultured. The sub-fusion cultured cells were divided into blank control group, Astragaloside IV group, UVA group and UVA + Astragaloside group and irradiated with UVA at 10 J / cm2 with 20 μg / mL Astragaloside intervention treatment. The expression of aging related β-galactosidase was detected by histochemical staining. The content of transforming growth factor-β1 (TGF-β1) in the supernatant was detected by enzyme-linked immunosorbent assay (ELISA). Real-time polymerase chain reaction Method to detect the mRNA expression level of MMP-1 and TIMP-1. Results: The β-galactosidase positive cells in the blank control group and astragaloside group were lower than those in the UVA group, the number of β-galactosidase positive cells in the UVA group was significantly increased. The addition of astragaloside treatment could make β-galactosidase positive cells The rate was significantly lower (P <0.05). Astragaloside treatment could promote the secretion of TGF-β1 by fibroblasts; the secretion of TGF-β1 by UVA irradiation was significantly decreased, and the addition of astragaloside IV before UVA irradiation could increase the secretion of TGF-β1 (P <0.05). In addition, UVA could induce the mRNA expression of MMP-1 and TIMP-1 to increase, while astragaloside could inhibit the expression of MMP-1mRNA and induce the expression of TIMP-1mRNA to a certain extent (P <0.05). CONCLUSION: Astragaloside IV can effectively delay the photoaging process of human fibroblasts, and its mechanism may be related to promoting the secretion of TGF and inhibiting the collagen degradation.