Study On Protective Effect And Mechanism Of Ursolic Acid And Kaempferol On UVB Damage In HaCaT Cells

Objective:To investigate fructus corni and dodder active ingredients of ursolic acid and campherol from Shijing Pill on ultraviolet?UVB?damage to human skin keratinocytes?HaCaT?protective effect and observe acid and campherol on UVB damage effect of HaCaT cell of TNF-??IL-6 related inflammatory factors and JNK?P-JNK?AKT?P-AKT signal transduction and regulation mechanism,in order to provide theoretical basis for the study of the prevention and cure of skin photoaging by Shijing Pill.Methods:The logarithmic growth phase HaCaT cells were uesd to establish light aging model by UVB irradiation at doses of 60mJ·cm-2.The cell proliferation rate were detected by MTT assay to determine the safe concentration of drugs and effective concentration.The different concentrations of Ursolic acid(10-14 mol·L^-1)and campherol(10-10 mol·L^-1)were uesd for photoaging HaCaT cells 24h,then detected superoxide dismutase?SOD?activity and malondialdehyde?MDA?content in the cells by the kit.The Western blot method was used to detect tumor necrosis factor a?TNF-???interleukin 6?IL-6??AKT?P-AKT?JNK?P-JNK and any other related indicators.Result:Cell morphology was observed under inverted microscope,normal HaCaT cells were arranged closely the boundary was clear.However,the gap of HaCaT cells became larger,the shape was irregular and the boundary was blurred.2.MTT results showed that when the concentration of ursolic acid was higher than 10-12mol and the concentration of campherol was higher than 10-8mmol will inhibit the normal proliferation of HaCaT?P<0.01,P<0.05?.In the safe concentration range,ursolic acid at the concentration of 10-14mo1·L^-1 and campherol at the concentration of 10-10mol·L^-1)protective effect on photoaging of HaCaT the most significant?P<0.01?.3.Compared with the blank group,the activity of SOD in model group was significantly decreased,and the content of MDA was significantly increased?P<0.01?.Compared with the model group,the concentrations of ursolic acid(10-14mol·L^-1)and campherol(10-10mol·L^-1)can significantly improve the intracellular SOD activity and Reduce MDA content?P<0.001?.Compared with the blank group,the protein expression of TNF-a and IL-6 and the ratio of P-AKT/AKT and P-JNK/JNK were significantly increased in the model group?P<0.01?;Compared with the model group,the concentrations of ursolic acid(10-l4mol·L^-1)and campherol(10-10mol·L^-1)can significantly decreased the protein expression of TNF-a and IL-6 and the ratio of P-AKT/AKT and P-JNK/JNK?P<0.01?.Conclusion:Ursolic acid and campherol could promote the proliferation of HaCaT cells in photoaging,increase the activity of SOD,decrease the content of MDA,inhibition the protein of TNF-? and IL-6 expression,so as to effectively prevent the occurrence of skin photoaging.