Studies On Purification,Analysis And Anti-inflammatory Activity Of Phenolics From Aerial Part Of Salvia Bowleyana Dunn

Salvia bowleyana Dunn,a perennial herb of genus Salvia,is mainly produced in Fujian and other southern provinces.Its root can be used as medicine,whose efficacy is the same as Chinese medicine S.miltiorrhiza.Early research had shown that its root is rich in four kinds of strong antioxidant activity phenolic acids,such as salvianolic acid B and rosmarinic acid.On this basis,the phenolic compounds from the aerial part(stem,leaf,inflorescence)of S.Bowleyana were purified and analyzed,and then main phenolic acids were carried out anti-inflammatory activity on the cell model.The result could provide scientific basis for the potential clinical application of phenolic acids and make full use of wild resources of S.bowleyana.The main results are as follows:(1)The extracts in root,stem,leaf and inflorescence of S.bowleyana were prepared by ultrasonic extraction-macroporous resin adsorption,their total phenol content were about 50%respectively.Nine fractions were isolated from the inflorescence extract.Using polyamide and Sephadex LH-20 as column chromatography medium,ten kinds of phenolic compounds—salvianolic acid B(C36H30O16),rosmarinic acid(C18H16O8),nepetoidin B(C117H14O6),salvianolic acid C(C26H2OO10),nepetoidin B dimmer(C34H28O12),kaempferol-3,7-di-O-rutanoside(C39H5OO24),kaempferol-3-?-D-(6"-O-malonyl)-glucopy-ranoside(C24H22O14),rutin(C27H30O16),isoquercitrin(C21H20O12),isorhamnetin(C16H12O7),were confirmed from above fractions by HPLC,LC-MS and LC-MS2.Five kinds of phenolic compounds-nepetoidin B,kaempferol-3,7-di-O-rutanoside,kaempferol-3-?-D-(6"-O-malonyl)-glucopyranoside,isoquercitrin and isorhamnetin were first isolated from S.bowleyana.(2)Using Sephadex LH-20 as chromatography medium,different volume fraction of ethanol solution as eluent,five fractions were obtained from the stem and leaf extracts respectively.Four kinds of phenolics-salvianolic acid B,rosmarinic acid,rutin and nepetoidin B were confirmed from purification fractions of stem and leaf by HPLC,LC-MS and LC-MS2.(3)Comparing the purified products of root,stem,leaf and inflorescence extracts of S.bowleyana,we finded that salvianolic acid B and rosmarinic acid were common components of different organs.The yield of salvianolic acid B was ranked as follows(in descending order):root(4.36%)>leaf(0.58%)>stem(0.37%)>inflorescence(0.11%),and the yield of rosmariinnic acid was ranked as follows:inflorescence(0.42%)>root(0.33%)>leaf(0.32%)>stem(0.14%).Therefore,salvianolic acid B was mainly stored in root,and the content of rosmarinic acid in inflorescence was higher then another organs,which provided a scientific basis for the selection of sources when this two components were prepared.(4)Using interleukin-6(IL-6)as detectable gene,the preliminary experiment with different concentrations and treatment time on RAW264.7 cell inflammatory model showed that 100?M and 12 h were optimal concentration and effect time of salvianolic acid B and rosmarinic acid.With pretreatment of 100 ?M salvianolic acid B and rosmarinic acid and treat 12 hours,we detected the mRNA expression levels of four kinds of inflammatory factors(IL-6,IL-1?,TNF-?,CXCL10)in the cell after 3 hours of LPS-induced.The results show that salvianolic acid B and rosmarinic acid had extremely significant inhibitory effects on four kinds of inflammatory factors(p<0.01),and the inhibitory effect of salvianolic acid B on IL-6 and IL-1?? was stronger than that of rosmarinic acid,while the inhibitory effect of rosmarinic acid on TNF-? and CXCL10 was stronger than that of salvianolic acid B.It suggested that salvianolic acid B and rosmarinic acid were good natural anti-inflammatory substances,which can provide more choices for the research and development of clinical medicine.Futher studies could examine their effects on MAPK,NF-?B and Akt signaling pathways,and then could reveal their anti-inflammatory mechanisms.