Protctive Effects Of Isopsoralen And PDG On UVB-induced HDF Cells Damage And Its Mechanism

Objective:To observe Isopsoralen and Pinoresinol Diglucoside(PDG)that are two plants of phytoestrogen from Psoralen and Eucommia,which have protective effects of the UVB damaging cells,and to explore protective mechanism of the Isopsoralen and PDG for ER pathway and downstream MAPK pathway on UVB damage cells,which phytoestrogens offer a theoretical basis for protection and treatment of photoaging.Methods:1.Construction of photoaging damage model,we selected exposure dose of 1min,2min,3min,4min,5min,1Omin,the cell proliferation rate were detected by MTT assay to determine the optimal dose of photodamage.2.The concentration of Isopsoralen and PDG(100μM,10μM,1μM,10-1M,10-2μM,10-4μM)were acted on HDF cells,the rate of cell proliferation was determined by MTT to determine the concentration of both drugs’effective and protection.3.The Western blot and real-time PCR method were used to detecte the expression protein of COL I and MMP-1;while adding estrogen receptor blockers(ICI182780)and P38 pathway inhibitor(SB203580)to detecte the expression protein of ERp,p38,p-p38 and expression mRNA of MMP-1.Results:1.Compared with the control group,UVB irradiation time(3min,4min,5min,1Omin)of cell proliferation rates were 92%,80%,69%,50%(p<0.01),the cells of irradiation time(5min)could be maintained to normal situation after treatment of phytoestrogens,in order to determine dose of UVB irradiation was 150mJ/cm2.2.Compared with the control group,the proliferation rate of psoralen(1OOμM)and PDG(100 μM)were significantly decreased(p<0.01),this concentration had apparently toxic effects on cells.Compared with the model group,the concentration 1μM,10-1 μM,10-2μM)of psoralen and PDG were significantly increased(p<0.01),in order to determine effective and protective concentrations.3.Compared with the model group,the concentration(10μM,1μM,10-1μM)of Isopsoralen and PDG could be significantly increased collagen protein expression(p<0.01),the concentration of Isopsoralen(10μM,1μM,10-1μM)and PDG(10μM)could be inhibited MMP-1 protein expression(p<0.01),Ipsoralen(10μM)and PDG(10μM)were significantly inhibited mRNA expression levels of MMP-1(p<0.01),while significantly were increased the expression of ERβ protein and decreased p-p38 protein expression(p<0.01);Compared with the phytoestrogens group,ICI182780(1μM)was significantly suppressed ERβ protein expression and up-regulated MMP-1 mRNA expression(p<0.01),while SB203580(10μM)significantly was decreased MMP-1,p-p38 protein expression and mRNA expression of MMP-1(p<0.01).Conclusion:we find that Isopsoralen and PDG can significantly promote the proliferation rate of HDF cells,promote collagen protein expression,suppress MMP-1 protein expression,up-regulate ERβ protein expression and inhibit the phosphorylation of p38 expression to inhibit degradation of MMP-1,further promote the synthesis of collagen,thereby slowing the incidence of skin photoaging.