Study On The Separation And Pharmacological Activities Of Middle And High Polar Layer Constituents From The Cissus Pteroclada Hayata

Cissus Pterolada Hayata is a genus of perennial vine plant belonging to the fam-ily Vi?taceae which is widely distributed in Guangxi,Hainan province of China.It have many pharmacological functions such as treated blood circulation promo-ting,eliminating inflammation,rheumatisme liminating and anti-inflammatory etc.In this paper,based on the raw material of Cissus Pteroclada Hayata study on the isolation and structure identification of high and medium polar layer constituents and the optimization of the extraction process of total polyphenols by Response Surface Methodology and the inhibitory effect and mechanism of the formation of melanin in murine B16 melanoma cells.1?25 compounds were isolated from high and medium polar layer by column chrom-atography,Prep-HPLC,and Medium pressure HPLC,and the structures of 21 compo-unds were identified,compound 3 and compound 5 were isolated from this plant for the first tnme.They are Stigxnasterol(1)??-sitosterol(2)>Laurie acid(3)?Taraxerone(4)?Imperatorin(5)?Ellagic acid(6)?Daucousterol(7)?oleanolic acid(8)?Resver atrol(9)?Polydatin(10)?11-O-galloylbergeni(12)?11-O-p-hydroxybenzoybergenin(13)?Bergenin(14)?(7E,9Z)-dihydrophaseie acid 3-O-?-D-glucopyranoside(15)?Ber-genin-11-O-?-D-galactopyranoside(16)?N-palmitie acid(19)?Gallic acid(20)?Ethyl gallate(21)?methyl gallate(22)?Myri-cetin(24)?Rutin(25).2?Using Response Surface Methodology to optimize the extraction process of total p olyphenols from the Cissus Pteroclada Hayata.Based on the single factor test,Response Surface Methodology test was carried out to investigate the effect of ethanol concentration,solid-liquid ratio,extraction time,extraction temperature on the extraction rate of total polyphenols,the extraction rate of total polyphenols from the Cissus Pteroclada Hayata was evaluated to find the best conditions.The optimum parameters of total polyphenols were:the concentration of ethanol was 80%,solid liquid ratio was 1:25,the extraction time was 2.4 h,the extraction temperature was 100?,the extraction rate of total polyphenols was 0.807%.The experimental results show that the Response Surface Methodology is reasonable and feasible to optimize the extracti-on conditions of total polyphenols from the Cissus Pteroclada Hayata,Provide a theoretieal basis for improving the extraction rate of total polyphenols3?Using murine B16 melanoma cells as a model,the proliferation rate of B16 melan-ma cells,the activity of tyrosinase in B16 melanoma cells,the content of melanin inB16 melanoma cells,the content of reactive oxygen species in B16 melanoma cells,the content of GSH in B16 melanoma cells was uesd as a basic evaluation standardsy to explore expression of tyrosinase in melanoma cells.The ethanol extract of Cissus Pteroclada Hayata al was the best drug to inhibition the generation of melanin in mouse B16 melanoma cells.