何首乌与制何首乌对人尿源性干细胞的作用差异及机制研究

目的 以人尿源性干细胞(human urine-derived stem cells,hUSCs)为研究对象,探究何首乌(Polygoni Multiflori Radix,PMR)与制何首乌(Polygoni Multiflori Radix Praeparata,PMRP)对人成体干细胞的作用差异及机制.方法 利用...

Study on the Differential Effects and Mechanism of Polygoni Multiflori Radix and Polygoni Multiflori Radix Praeparata on Human Urine-derived Stem Cells

Objective Human urine-derived stem cells(hUSCs)were used to explore the differentiation effect and mechanism of Polygoni Multiflori Radix(PMR)and Polygoni Multiflori Radix Praeparata(PMRP)on human adult stem cells.Methods Quantitative analysis of Emodin(EM)and 2,3,5,4’-tetrahydroxystilbene-2-O-β-D-glycoside(TSG)in the aqueous extract of PMR and PMRP was carried out by high Performance Liquid Chromatography;MTT assay was used to study the influence of PMR and PMRP on hUSCs viability;according to the content and proportion of EM and TSG in the two aqueous extracts,the effects of single component and 2 components combined in proportion on hUSCs viability were compared;cell cycle distribution and cell apoptosis were analyzed by f low cytometry,and cell cyclerelated proteins were detected by Western blot.Results The contents of TSG and EM in PMR were 77.68 and 0.53 mg/g,and were 29.37 and 0.36 mg/g in PMRP.PMR had obvious toxicity to cells at 0.5 and 1.0 mg/mL,while PMRP had the effect of promoting cell proliferation at the same dose.EM promoted cell proliferation at 0.5~4μmol/L,and TSG inhibited cell activity at 320 and 640μmol/L.The inhibitory effect of the combination of EM and TSG on cell viability was significantly increased compared with the corresponding concentration alone.The G0/G1 cell cycle arrest occurred in the PMR group and the 1∶160 combined group,and the cell apoptosis rates were higher than the PMRP group and the 1∶80 combined group.Western blot results showed that p21 level was upregulated in the PMR group.CDK1,CDK2 and Rb phosphorylation level were down-regulated in the PMR group,while up-regulated in the PMRP group.Conclusion Under the same conditions,PMR showed obvious cytotoxic effect compared with PMRP,while PMRP showed the effect of promoting cell activity,indicating that the change of EM and TSG proportion may be one of the mechanisms of the PMR processed attenuating toxicity.