Penetration Promotion Mechanism And Skin Whitening Effect Of Ligusticum Jeholense Nakai Et Kitag

Ligusticum jeholense Nakai et Kitag,a member of the Umbelliferae plant,has been used as the traditional medicine Gao-Ben（GB）in China.The rhizome and root of GB have warm properties as well as spicy flavours and possess efficacies in promoting qi and blood circulation,expelling wind,and alleviating pain.GB has a long history of medicinal use in China,the earliest dating back to Sheng Nong’s Herbal Classic（Qin and Han Dynasty,around 221 B.C.to 220 A.D.）.GB is widely distributed in northeast China.Traditional Chinese medicine（TCM）has a long history of skin whitening use inChina,the earliest dating back to the Jin Dynasty（around 317-420 A.D.）.It was initially used as a deodorant to improve body scent in A Handbook of Prescriptions for Emergencies and was then widely used in external prescriptions until the Tang Dynasty（618-907 A.D.）,the strongest and most flourishing era in the history of China.With social prosperity and scientific as well as technological progress,TCM for external use achieved rapid development during this period.GB has been widely used in external prescriptions for skin whitening in TCM.Considering that natural plant extracts are increasingly used in cosmetic products,the aim of the study was to investigate the functions of GB in topical application prescriptions for skin whitening.Firstly,this paper analyzed the prescription rules of skin whitening formulas containing GB by TCM Inheritance Support System platform（V2.5）.There were 97 prescriptions contained GB.To understand whether the GB is only a penetration enhancer or has the function of skin whitening in beauty prescriptions,the following work was mainly done in this paper:1.The supercritical CO₂ Fluid extraction（SFE-CO₂）process was optimized by single factor experiment and response surface method.The SFE-CO₂ extraction conditions of lipid-soluble constituents in GB（GBO）were determined.The optimal extraction temperature of GBO was 53℃,the extraction pressure was 35 MPa,the extraction time was 110 min,and the CO₂ flow rate was 20 L/h.The average yield of GBO could reach more than 10%.2.The chemical composition of GBO was determined by gas chromatography-mass spectrometry（GC-MS）,and a total of 32 compounds were identified.The two substances with the highest content were Benzyl Alcohol（55.29%）and Ligustilide（35.68%）.3.To study the effects of GBO on the transdermal amount of drugs,in vitro transdermal tests were performed using the Franz diffusion cell.The cumulative amount of model drugs that penetrated through the tested skins was quantified by high-performance liquid chromatography.The results of the Franz diffusion cell experiment showed that GBO at 1%～5%concentrations did have the effect of promoting penetration.4.The changes in morphological features of the skin treated with GBO were identified by computer-aided image analysis of haematoxylin and eosin staining.The penetration-promotion mechanism of GBO was increasing skin permeability by impacting skin structure,thinning out the stratum corneum and losing the epidermal structure.The results showed that the 5%GBO group and 5%azone group had similar effects on the structure of the stratum corneum.5.Through the network pharmacology of TCM,the main ingredient of skin whitening were linoleic acid,ligustilide,coumarin,and hyperoside.The main targets involved were IL6,TNF,JUN,ALB,MAPK3,EGFR,STAT3,and PTGS2.KEGG enrichment analysis results mainly included pathways in cancer,the AGE-RAGE signalling pathway in diabetic complications,proteoglycans in cancer,and HTLV-I infection.6.Molecular docking further demonstrated that the active ingredients,such as linoleic acid,ligustilide,coumarin,hyperoside,paeonol,β-sitosterol,and other whitening targets,such as TNF,TYR,PTGS2,MAPK3,had a good binding effect.