衰老HDF细胞DNA损伤与HSP70mRNA表达及山茱萸多糖的作用

Fructus corni polyaccharides against DNA damage and effect on the expression of heat shock protein 70 mRNA

目的探讨山茱萸多糖(FCP)血清对抗人胚肺二倍体成纤维(HDF)细胞衰老的作用及热休克蛋白(HSP)70的调控机制。方法体外培养HDF细胞。根据HDF细胞代龄和施加因素分为4组,即青年组(32±3代)、多糖组(52±3代用1%多糖含药兔血清处理)、对照组(52±3代用1%正常兔血清处理)和衰老组(52±3代)。自45代开始加含多糖药血清并以空白血清培养作对照。观察细胞形态,MTT检测细胞活力,单细胞凝胶电泳检测DNA损伤,RT-PCR检测HSP70mRNA表达。结果与青年组比较,衰老组细胞活力明显下降(P<0.01),DNA损伤率增加(P<0.01),HSP70mRNA表达下降(P<0.01);多糖组细胞活力明显增高(P<0.01),DNA损伤下降(P<0.01),HSP70mRNA表达增加(P<0.01)。结论FCP血清可以通过调节HSP70的表达减轻细胞DNA损伤从而发挥抗HDF细胞衰老的作用。

Objective To investigate the anti-aging effect of the serum with Fructus Corni polysaccharides（FCP） and the possible mechanism of heat shock protein70 on human diploid fibroblasts（HDF） aging cell.Methods Conventional culture was performed in the HDF cells,the serum of FCP and control blood serum were added to the experiment group from 45 generation.The cell changes were observed under microscope.The cell activity was detected by MTT method.The DNA damage was detected using single cell gel electrophoresis（SCGE）.RT-PCR method was used to examine the expression of HSP70mRNA.Results In aging group the cell activity was decreased（P0.01） and the levels of DNA damage were increased （P0.01）, the expression of HSP70 mRNA were depressed（P0.01）.Compared with the aging group,the cell activity and the expression of HSP70 mRNA were increased（P0.01）,the levels of DNA damage were reduced in FCP group.Conclusions The possible mechanism of anti-aging effect of the serum FCP is to regulate the expression of HSP70 mRNA,to reduce the DNA damage.