Study On The Effect Of The Extract Of Polygonum Multiflorum And Its Main Monomer Components On Tyrosinase And Antioxidant Activity

Chinese medicine polygonum multiflorum thumb is the root of the perennial herbaceous plant of polygonaceae.Research has shown that polygonum multiflorum Processed make the physiologically active ingredients significantly change,Changes in its efficacy may be related to the content of chemical composition before and after the processing.The hair blacking effects of polygonum multiflorum processed whether it is because after the processing some content has changed,and what ingredients is related and the mechanism of hair blacking remain to be studied to determine.Objective:To explore the separate part of the polygonum multiflorum processed and the main monomer component changed of content before and after the processing effect on tyrosinase activity and mRNA expression levels.To explore if the PWE and the main monomer compositions of processed Ploygonum multiflorum(emodin,THSG)can improve the anti-oxidation damage ability of cells through Nrf2-ARE pathway,Surmise if the potential ability of processed Ploygonum multiflorum can protect the melanin melanin/stem cells from oxidative damage.Methods:Selecting various the separate part of the polygonum multiflorum processed and the main monomer component changed of content before and after the processing to make experiments with tyrosinase activity impact extracellularly by the rate oxidation of dopa assay.Through the rate oxidation of dopa and dopa staining testing to detect the separate part of the polygonum multiflorum processed impact to tyrosinase activity of B16F10;detecting the polygonum multiflorum processed impact on the tyrosinase mRNA expression of B16F10 by RT-PCR.By using the method of DPPH radical scavenging assay to determind the antioxidant capacity of the separate part of the polygonum multiflorum processed and the main monomer component of the extracellular.Detecting the separate part of the polygonum multiflorum processed and the main monomer component impact on ARE(antioxidant response element)report gene expression on HepG2-ARE by luciferase assay.Results:Significant activation of tyrosinase activity component did not be screened from the test drug,but the water extract of polygonum multiflorum processed(PWE),ethanol extract of polygonum multiflorum processed inside the 50%ethanol eluate and 95%ethanol eluate,2,3,5,4 tetrahydroxy stilbene-2-O-?-D-glucoside(THSG),emodin and so on showed some inhibition of tyrosinase activity.By detecting the effects of PWE,polygonum multiflorum processed polysaccharide and THSG,Emodin on.tyrosinase activity on intracellular,we discovered the ingredients inhibited the activity of tyrosinase in varying degrees,which was consistent with the results of activity affect extracellular,these ingredients of polygonum multiflorum processed in the concentration range tested showed no significant effect on the expression levels of tyrosinase.Besides,we found that PWE and the THSG can play the remarkable antioxidant effect investigated via DPPH radical scavenging assay.By detecting the ARE expression in HepG2-ARE cells we found that PWE in the concentration range of 100?400 ?g/ml and the THSG in the concentration range of 15?60?g/ml can activate the expression of ARE and promote the antioxidant effect in a concentration-dependent manner,and the emodin at low concentrations can also present certain antioxidant effect.Conclusion:The results cannot support the opinion that polygonum multiflorum processed directly activate tyrosinase activity to black hair by the selected test components;Whether the existence of the other ingredients or the ingredient of polygonum multiflorum processed after metabolismcan in vivo can activate tyrosinase activity and expression remained to be studied;In addition,the results showed the processed ploygonum multiforum can protect cells from oxidative damage by activation of Nrf2-ARE pathway and delay aging,but whether the processed ploygonum multiflorum can actually protect melanin melanin/stem cells from oxidative damage and the specific mechanism needs further study.