Protective Effects Of Baicalein On UVB Induced Photoaging In HaCaT

Objective:To observe baicalein that is plant of flavonoids from Eucommia,which have protective effects of the UVB damaging cells,and to explore protective mechanism of the baicalein for MAPK and NF-κB pathway on UVB damage cells,which baicalein offer a theoretical basis for protection and treatment of photoaging.Methods:1.The effects of baicalein on the proliferation of normal HaCaT cellsThis experiment uses the different concentrations of baicalein,and determined by MTT method is used to determine on HaCaT cells,hence the best effective concentration screening drugs,including the minimum concentration of 10-11 mol/L,the remaining concentration as the minimum concentration in turn 10 times,100 times,1000 times,10000 times,100000 times,10 million times and 100 million times.2.Establish the photoaging HaCaT cell modelChoosing radiation intensity of 0.5mW/cm2 UVB to illuminate,which respectively select the(1,3,5,7,10,15)min irradiation time,the cell proliferation rate were detected by MTT assay to determine the optimal dose of photodamage.3.Baicalein has an effect on the proliferation and morphology of photoageing HaCaT cellsUsing three concentrations of baicalein light aging cells,concentration respectively the concentration of 10000 times,100000 times and 10000 times as much as three concentrations,and determined by MTT method is used to determine cell proliferation rate,in scutellaria baicalensis were observed under inverted microscope light aging changes in cell morphology.4.Baicalein affects the levels of SOD,GSH,CAT activity,and MDA and ROS in the cells of photoaging HaCaT cellsTest kits were used to detect the effects of SOD,GSH,CAT activity,and the changes of MDA and ROS in photoaging cells.5.The RT-PCR method tests the effects of the expression of IL-la,I1-8,and TNF-αmRNA in photoageing HaCaT cellsThe expression level of IL-1α,Il-8,TNF-α mRNA in photoageing cells was tested by RT-PCR.6.WB method tests the effects of the expression of the expression of the expression of IL-1α,I1-8,TNF-α,p-ERK1/2,ERK1/2,p-P38,P38,NF-κBP50,NF-κBP65 protein in photoageing HaCaT cellsThe WB method was used to detect the expression of p-ERK/ERK,p-P38/P38,NF-κBP50/NF-κBP65,IL-1α,Il-8,TNF-α protein in photoaging cells in photoageing cells.Results:1.Baicalein has an effect on the proliferation of normal HaCaT cellsCompared with the blank group,(10-11,10-10,10-9)mol/L three concentration group and 10-8 mol/L concentration cell proliferation rate increased significantly(p<0.05,p<0.01),the concentration of low concentration of two groups of significant effects on the cell increment rate,the difference was statistically significant(p<0.01).There was no significant effect on the cell proliferation rate of the three concentrations in the concentration group,no statistical difference(p>0.05),to determine the optimum concentration.2.UVB screening of HaCaT cell exposure timeCompared with the blank group,the control group of UVB(7min)and the control group of UVB(10,15min)had significant influence on cell proliferation rate,and the difference was statistically significant(p<0.05,p<0.01).Compared with the control group in the last three groups,the control group of the first three groups(1,3,5min)had no significant effect on cell proliferation rate,no statistical difference(p>0.05).3.Baicalein has an effect on the proliferation and morphology of photoageing HaCaT cellsCompared with the blank group,model group had no significant role in promoting cell proliferation of HaCaT cells(p>0.05);Compared with model group,(10-7,10-6,10-5)mol/L baicalein group had no significant role in promoting proliferation of HaCaT cells(p>0.05);Under the microscope,the blank group cells are rhombic,plump,and normal.Compared with the blank group,the cells in the model were shrivelled and oval-shaped,and some of the cells in the cell were damaged and tended to die.Compared with model group,10-7mol/L baicalein group reduce cell damage phenomenon,10-6mol/L baicalein group cell shrinkage and significantly reduce breakage phenomenon,10-5mol/L baicalein group of cells in good condition,the cell body was close to normal.4.Baicalein affects the levels of SOD,GSH,CAT activity,and MDA and ROS in the cells of photoaging HaCaT cellsCompared with the blank group,the activity of SOD,GSH and CAT decreased,the content of MDA and ROS increased in model group(p<0.05,p<0.01).Compared with model group,(10-7,10-6,10-5)mol/L baicalein group significantly improve light aging cells,the activity of SOD,GSH and CAT increased,the content of MDA and ROS decreased(p<0.05,p<0.01).5.Baicalein affects the expression of IL-1α,IL-8,TNF-α mRNA in photoageing HaCaT cellsCompared with the blank group,IL-1α,IL-8 and TNF-α expression of quantity of mRNA increased in model group(p<0.05,p<0.01).Compared with model group,(10-7,10-6,10-5)mol/L baicalein group significantly decreased light aging cells of IL-1α,IL-8 and TNF-α expression of quantity of mRNA(p<0.05,p<0.01).6.Baicalein affects the expression of IL-1α,IL-8,TNF-α,p-ERK,p-P38,NF-κB P50,NF-κBP65 protein in photoageing HaCaT cellsCompared with the blank group,IL-1α,IL-8,TNF-α,p-ERK,p-P38,NF-κBP50,NF-κBP65 expression of protein significantly increased in model group(p<0.05,p<0.01).Compared with model group,(10-7,10-6,10-5)mol/L baicalein group significantly decreased light aging cells of IL-1α,IL-8,TNF-α,p-ERK,p-P38,NF-κBP50,NF-κBP65 expression of protein(p<0.05,p<0.01).Conclusion:Baicalein can significantly promote protect HaCaT cells light aging,improved the activity of SOD,GSH and CAT,reduced the content of MDA and ROS,decreased light aging cells of IL-1α,IL-8 and TNF-α expression of q uantity of mRNA and protein.Also can regulate phosphorylation p-ERK1/2/ER K1/2,p-P38/P38,NF-κBP50/NF-κBP65 expression of protein,thereby inhibiting L-1α,IL-8 and TNF-α,slowing the incidence of skin photoaging.