Protective Effect Of Myricaria Paniculata Compounds And Essential Oil On HaCat Cell Damage Caused By Ultraviolet And UVB

Tibetan medicine Myricaria paniculata is a twig of Myricaria paniculata,a plant belonging to the genus Myricaria of the family Tamaricaceae,it mainly treats plague epidemics,and viscera poisonous heat,sore throat and yellow water disease etc.The research group obtained 24 compounds and essential oils from Myricaria paniculata in the early stage and carried out biological activity analysis.The results showed that Myricaria paniculata compounds and essential oils have good antibacterial,anti-inflammatory,antipyretic,and analgesic pharmacological activities,while the compounds and essential oils have little research on skin aging damage caused by ultraviolet radiation.Based on the previous research,this paper explored the protective effects of 7 active ingredients and essential oils on the damage of HaCat cells caused by ultraviolet and UVB.The main research results are as follows:1.A model of HaCat cells irradiated by UV-UVB was established.After 5,10,15,and 20 min of UVB irradiation at 8 m J/cm~2,the cell viability was determined by CCK-8.The results showed that the survival rate of cells decreased to below 50%after 20 min of irradiation,and the survival rate of cells was 83%after 10 min of irradiation.Therefore,UVB irradiation was used for 10 min in subsequent experiments,and the total irradiation dose was 24 J/cm~2to establish a UVB cell model.2.The CCK-8 method was used to determine the effects of different concentrations of 7 compounds and essential oils on HaCat cells at different times,thereby affecting the viability of HaCat cells,compound groups were treated with 3,5,and 10μmol/L for 4,6 and 8 hours,respectively.The results showed that different concentrations were used for 4 h,except for compound 1 and compound 7,the cell viability of the other compounds was the highest at 3μmol/L.The maximum cell viability was over 100%when the concentration was 3μmol/L at 6 h and 8 h,and 10μmol/L and above had an inhibitory effect on the cells,indicating that the high concentration of the compound did not further enhance the cell viability;Under the same concentration and different time,the cell viability reached the maximum for 6 h after the compound acted on it.In the Myricaria paniculata Essential oil group,the concentration of 0.01,0.1,0.5%,and 1%v/v was respectively used for 4,6,8,and 10h,The results showed that the essential oil concentration was 0.01,0.1 and 0.5%v/v,and the cell viability showed an upward trend,reaching the maximum value after 8 h.Therefore,the subsequent experimental compound group was treated with 3μmol/L for 6 h,and the essential oil group was treated with 0.01,0.1 and 0.5%v/v for 8 h.3.Use UVB irradiated HaCat cell model.Compounds and essential oils were used on HaCat cells before irradiation,followed by irradiation.After 24 h of culture,the contents of SOD,MDA,ROS,and related inflammatory factors TNF-α,IL-6,and Caspase-3 were measured.The results showed that there were significant differences between the Myricaria paniculata 7 compounds treatment group and Myricaria paniculata essential oil treatment group and the model group（P<0.05）.The results indicated that 7 compounds of Myricaria paniculata and Myricaria paniculata essential oil could improve the changes of cell morphology caused by UVB irradiation of HaCat cells and maintain cell viability.It can also effectively reduce the content of IL-6,TNF-αinflammatory factors and Caspase-3 protein,and each treatment group and model group、model group and normal group all reached extremely significant levels（P<0.01）.It shows that the compounds and essential oils have good anti-inflammatory effects.It can effectively inhibit Caspase-3 protein,and play an anti-photoaging effect.4.Network pharmacology and molecular docking analysis of 7 compounds in Myricaria paniculata and 10 active ingredients in essential oils,The mechanism of action of Myricaria paniculata compounds and essential oils in inhibiting the apoptotic damage induced by UVB irradiation in HaCat cells was predicted.By constructing a"drug ingredient-target-pathway"map,9 compounds,53 targets,and 20pathways were screened,which could further provide a reference for the study of the mechanism of treating skin inflammatory diseases.The 5 active ingredients and 4 key proteins obtained by molecular docking were verified,which showed that the active ingredients of Myricaria paniculata compound and Myricaria paniculata essential oil play a role in improving skin inflammation through the key targets of regulation.