Establishment And Application Of The Evaluation Method For Soothing Efficacy Of Cosmetic Ingredients

Research Background: Skin irritation refers to a reaction of the skin when it is irritated by the action of irritating substances,which symptoms as redness,swelling,tingling,itching and so on.In recent years,the diversification of cosmetics and cosmetic ingredients has greatly increased the possibility of inducing skin irritation,and people are more and more concerned about skin care and increased understanding of skin irritation,which led to a large demand in cosmetics and cosmetic ingredients that claim with soothing efficacy.Now,a lot of laboratory methods for evaluating soothing efficacy are based on inflammation,but skin irritation is a complex process.Anti-inflammation is an important indicator for evaluating the soothing efficacy of cosmetic ingredients,but is not the only evaluation indicator.Therefore,this thesis will develop a comprehensive,rapid,efficient and non-inflammation dependent method for evaluating the soothing efficacy of the ingredients.Research contents:1.An in vivo zebrafish embryos irritation model had been established.In this part,we used 24 hpf zebrafish embryos line AB to establish an irritation model.The irritation of different concentrations of SDS and 4-BR were evaluated by this model.Spontaneous rotating of 24 hpf zebrafish embryos was observed and recorded under the microscope with imaging system.We established a stable zebrafish embryo irritation model.2.Evaluation of soothing efficacy of cosmetic ingredients by zebrafish embryos irritation.The 24 hpf zebrafish embryos line AB were treated with different concentrations of dipotassium glycyrrhizinate,baicalein,?-bisabolol,paeonol,asiaticoside,proantho cyanidins,4-tert-butylcyclohexanol and tea polyphenols.The no observable effect concentration(NOEC)of the ingredients was determined according to the spontaneous rotating and death of zebrafish embryos.Then we used the zebrafish embryos irritation model to evaluate of eight ingredients and the soothing efficacy of the ingredients was determined based on the spontaneous rotating of zebrafish embryos.3.Study on soothing efficacy of cosmetic soothing ingredients.In order to establish a zebrafish inflammation model,the zebrafish line Tg[lyz: Ds Red] were treated with 100 ?mol/L SDS.Evaluation of anti-inflammatory efficacy of dipotassium glycyrrhizinate,baicalein and4-tert-butylcyclohexanol by zebrafish inflammation model.We used 24 hpf zebrafish embryos line AB to establish an irritation model,treated by dipotassium glycyrrhizinate,baicalein and4-tert-butylcyclohexanol,RNA samples were prepared at 24 hpf,and then the m RNA expression of relevant inflammatory factors was detected by RT-q PCR.Then we explored the relationship between the investigation of soothing efficacy of cosmetic ingredients and inflammatory factors.Research results:1.An in vivo zebrafish embryos irritation model had been established.Compare with the blank control group,the spontaneous rotating of zebrafish embryos was significantly enhanced when 500 ?mol/L SDS was applied for 15 min,indicating that SDS has a remarkable irritation effect under this condition.In summary,we successfully established an in vivo zebrafish embryos irritation model.2.Evaluation of soothing efficacy of cosmetic ingredients by zebrafish embryos irritation.We found that the NOEC of dipotassium glycyrrhizinate,baicalein,?-bisabolol,paeonol,asiaticoside,proantho cyanidins,4-tert-butylcyclohexanol and tea polyphenols at 1000?mol/L,100 ?mol/L,50 ?mol/L,200 ?mol/L,500 ?mol/L,1000 ?mol/L,150 ?mol/L and 3g/L.By this method,?-bisabolol,paeonol,asiaticoside,proantho cyanidins,and tea polyphenols performed soothing efficacy at high concentration.Dipotassium glycyrrhizinate,baicalein and 4-tert-butylcyclohexanol performed soothing efficacy at low,medium and high concentration.In conclusion,a soothing efficacy method had been established.The soothing efficacy of dipotassium glycyrrhizinate,baicalin and 4-tert-butylcyclohexanol were evaluated by this method.We further explore anti-inflammatory efficacy by traditional inflammation model.3.Study on soothing efficacy of cosmetic soothing ingredients.Dipotassium glycyrrhizinate,baicalein performed remarkable anti-inflammatory efficacy at 500 ?mol/L,1000 ?mol/L and 50 ?mol/L.However,4-tert-butylcyclohexanol at 200 ?mol/L,250 ?mol/L and 300 ?mol/L without anti-inflammatory efficacy.The results of RT-q PCR,IL-6 and TNF-? in 24 hpf zebrafish embryos showed no significant changes after treated by 500 ?mol/L SDS,and IL-6 and TNF-? in 24 hpf zebrafish embryos also showed no significant changes after treated by dipotassium glycyrrhizinate,baicalein and 4-tert-butylcyclohexanol.In summary,cosmetic ingredients did not achieve a soothing efficacy by inhibiting inflammation in a zebrafish embryo irritation model.Conclusion: An evaluation method of soothing efficacy had been established in this thesis by spontaneous rotating of 24 hpf zebrafish embryos.We used this method to evaluate the soothing efficacy of cosmetic ingredients.And initially verified that the soothing efficay of cosmetic ingredients is not necessarily inflammation-dependent,this method is to achieve the soothing efficacy by anti-irritation.Therefore,in this thesis the evaluation of soothing efficacy of cosmetic ingredients can be used to select the soothing ingredients with anti-inflammatory effects and without anti-inflammatory effects,This method will provide a rapid,efficient and high-throughput method for the future development of new cosmetic ingredients by the spontaneous rotating of zebrafish embryos.