Heat-treating Process,Phenolic Acids Composition Analysis And Anti-inflammatory Mechanism Research Of Salvia Miltiorrhiza Bunge Var.Alba Wu And Li,Miss.

Salvia miltiorrhiza Bunge var.alba Wu and Li,miss.,as the white variant of Salvia miltiorrhiza Bunge,is native to Laiwu,Zhangqiu and Tai’an in Shandong province.The roots of S.miltiorrhiza Bge.var.alba are value for the treatment of cardiovascular diseases,such as coronary heart disease,angina,hypertension,hyperlipidemia and stroke;In particularly,its roots have also been used to treat patients with thrombosis angiitis obliteranstype(TAO)in the local.As a new species,it is included in "standard of Chinese medicinal material in Shandong province" in 2002.The main chemical constituents of the roots of S.miltiorrhiza Bge.var.alba are both tanshinones and phenolic acids,as those in the roots of S.miltiorrhiza Bge..In them,the content of salvianolic acid B(SalB)is abundant,but salvianolic acid A(SalA)is very poor.However,many researches revealed that SalA has an excellent biological activity.Firstly,we determined the content of fenolic acids in S.miltiorrhiza Bge.var.alba by HPLC methods,founding that SalA cannot be detected.Then,the study was focused on the conversion of salvianolic acid B into salvianolic acid A in the tissue of the roots of S.miltiorrhiza Bge.var.alba.The conditions,such as pH values of aqueous solutions to wet the powder of the roots,reaction temperatures and reaction times for converting SalB into SalA,were investigated.The results indicated that the amount of produced SalA,in the cases of the powder wetted by a solution with pH 1.0,and treated at 120 ℃ for 4 h,is the largest than that treated with other conditions.In treated powder of the roots of S.miltiorrhiza Bge.var.alba,SalB was mainly converted into SalA,and that a little of other phenolic acids were produced.The phenolic acids in the treated powder were analyzed by LC-MS methods.SalA in it was also measured by HPLC methods.Then,seven known phenolic acid compounds were isolated and elucidated by spectroscopic analysis from the treated powder of the roots,named as:methl 3,4-dihydroxy-phenyl lactate,ferulic acid,rosmarinicacid-3-O-glucoside,rosmarinic acid,isosalvianolic acid A,salvianolic acid A and salvianolic acid C,respectively.Among them,methl 3,4-dihydroxy-phenyl lactate,ferulic acid,isosalvianolic acid A,salvianolic acid A and salvianolic acid C were newly produced during a period of time when the powder of the roots were treated.The anti-inflammatory effects of salvianolic acid A,salvianolic acid C and rosmarinic acid on human aortic endothelial cells(HAECs)induced by tumor necrosis factor alpha(TNF-a)were investigated by the experiments,such as BCECF/AM labeling assay was used to detect the adhesion ability of U937 cells to HAECs;DCFH-DA method to detect the production of intracellular ROS;The mRNA levels of cell adhesion molecule-1(ICAM-1),vascular cell adhesion molecule-1(VCAM-1),human interleukin-6(IL-6),monocyte chemoattractant protein-1(MCP-1),RAGE,P-selectin and nuclear factor kappa B(NF-κB)were detected by RT-PCR;IL-6 and MCP-1 protein levels were measured with ELISA;ICAM-1,VCAM-1,RAGE,P-selectin and NF-κB protein levels were measured with western blot(WB).The results showed that the three phenolic acids compounds could inhibit the inflammatory cytokines in HAECs induced by TNF-a,and effectively inhibit the production of ROS and U937 cells adhesion to HAECs.So,these phenolic acids compounds have good anti-inflammatory activity.