| Objective:To investigate the protective effects and the mechanism of liquir itin that from the Chinese herbal active ingredients of licorice on photoaging h uman keratinocytes(HaCaT)induced by ultraviolet B(UVB),which provided a theoretical basis for the prevention and treatment of skin photoaging.Methods:1.The different concentrations of liquiritin were used on HaCaT cells,the cell proliferat rate were detected by methyl thiazolyl tetrazolium(MTT)method to determine the best safe and effective concentration.2.The photoaging model was set up by the irradiation of UVB at dose of 30mJ/cm2,and the best safe and effective concentration were used on photoagi ng model HaCaT cells,MTT method was used to detect the effect of liquiritin on photoaging model HaCaT cells proliferation rate.3.The activity of superoxide dismutase(SOD),glutathione(GSH),catalase(CAT)and the content of malondialdehyde(MDA),reactive oxygen species(ROS)we re detected by the corresponding kits.4.Western blot method was used to detect the expression of interleukin-1β(IL,1β),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),B-cell lymphoma 2(Bcl-2),Bcl-2 Associated X Protein(Bax),Cysteinyl aspartate 3(Caspase-3)and Cy steinyl aspartate 9(Caspase-9)protein in photoaging cells.5.The effects of liquiritin on the total apoptosis of photoaging HaCaT cells were respectively measured by flow cytometry.6.The expression of Bcl-2,Bax,Caspase-3 and Caspase-9 mRNA in photoagi ng cells were measured by Real-time Quantitative PCR(RT-PCR).Results:1.Compared with the blank control group,the cell proliferation rate of the(10-10,10-9,10-8)mol/L liquiritin group were significantly increased(P<0.05,P<0.01),cell proliferation was obviously promoted;the cell proliferation rate of the(10-4,10-3)mol/L liquiritin group were significantly decreased(P<0.01),cell prolifer ation was obviously inhibited;(10-7,10-6,10-5)mol/L liquiritin group had no statis tical difference on cell proliferation rate(P>0.05),and had no obvious promoting or inhibiting effects on cell proliferation,as they were the best safe and effecti ve concentrations.2.Compared with the blank control group,the cell proliferation rate of the UVB group were significantly decreased(P<0.01);Compared with the UVB grou p,the cell proliferation rate of the(1×10-7,1×10-6,1×10-5)mol/L liquiritin+UVB group were significantly increased(P<0.01).3.Compared with the blank control group,the activity of SOD,GSH and CA T were significantly decreased in UVB group(P<0.01),the content of MDA and ROS were significantly increased in UVB group(P<0.01);Compared with the UVB group,the activity of SOD,GSH and CAT were significantly increased in(1×10-7,1×10-6,1×10-5)mol/L liquiritin+UVB group(P<0.05,P<0.01),the content of MDA and ROS were significantly decreased in(1×10-7,1×10-6,1×10-5)mol/L liquiritin+UVB group(P<0.01).4.Compared with the blank control group,the expression of IL-1β,IL-6 and TNF-a protein were significantly increased in UVB group(P<0.01);Compared with the UVB group,the expression of IL-1β,IL-6 and TNF-a protein were sig nificantly decreased in(1×10-7,1 1×10-6,1×10-5)mol/L liquiritin+UVB group(P<0.05,P<0.01).5.Compared with the blank control group,the total apoptosis rate of cells was significantly increased in UVB group(P<0.01);Compared with the UVB gro up,the total apoptosis rate of cells was significantly decreased in(1×10-7,1×10-6,1×10-5)mol/L liquiritin+UVB group(P<0.01).6.Compared with the blank control group,the expression level of Bcl-2 mR NA was significantly decreased in UVB group(P<0.01),the expression level of Bax,Caspase-3 and Caspase-9 mRNA were significantly increased in UVB grou p(P<0.01);Compared with the UVB group,the expression level of Bcl-2 mRNA was significantly increased in(1×10-7,1×10-6,1×10-5)mol/L liquiritin+UVB gro up(P<0.05,P<0.01),the expression level of Bax,Caspase-3 and Caspase-9 mRNA were significantly decreased in(1×10-7,1×10-6,1×10-5)mol/L liquiritin+UVB group(P<0.05,P<0.01).7.Compared with the blank control group,the expression of Bcl-2 protein was significantly decreased in UVB group(P<0.01),the expression of Bax,Caspas e-3 and Caspase-9 protein were significantly increased in UVB group(P<0.01);Compared with the UVB group,the expression of Bcl-2 protein was significantl y increased in(1×10-7,1×10-6,1×10-5)mol/L liquiritin+UVB group(P<0.05,P<0.01),the expression of Bax,Caspase-3 and Caspase-9 protein were significantly d ecreased in(1×10-7,1×10-6,1×10-5)mol/L liquiritin+UVB group(P<0.05,P<0.01).Conclusion:Liquiritin could promote the proliferation of photoaging HaCa T cells,increase the activity of SOD,GSH and CAT,decrease the content of MD A and ROS,inhibite the expression of IL-1β,IL-6 and TNF-α protein,decrease th e total apoptosis of photoaging HaCaT cells induced by UVB.In the meantime,I iquiritin could promote the expression of anti-apoptotic factor Bcl-2 mRNA and protein,inhibite the expression of the apoptosis-related factors Bax,Caspase-3 a nd Caspase-9 mRNA and protein,so as to effcctively inhibite the photoaging of the skin. |
PDF Full Text Request