Isolation And Its Anti-inflammatory Activity Of Salvia Miltiorrhiza Water-soluble Compounds

Salviae miltiorrhizae radix et rhizoma(SMRR)is the dried root and rhizome of Salvia miltiorrhiza Bge,which have effects of promoting blood circulation and removing blood stasis,promoting menstruation and relieving pain,cooling blood and eliminating carbuncle.Our previous studies have shown that the compound preparations consisting of SMRR,Rhizoma coptidis,forsythia,and safflower have significant antibacterial and anti-inflammatory effects.Further studies have shown that the fat-soluble components of Salvia miltiorrhiza have antibacterial activity.On this basis,this experiment studied the separation and anti-inflammatory effects of the water-soluble components of SMRR.The following results were obtained:(1)Isolation and identification of the water-soluble extract of SMRR : The water-soluble extracts of SMRR were isolated and purified by D101 macroporous resin gradient elution and silica gel column chromatography.The chemical properties and nuclear magnetic resonance spectra were analyzed and identified as: salvianolic acid B,rosmarinic acid,caffeic acid,protocatechualdenyde.(2)The effect of water-soluble extract of SMRR on ear swelling caused by xylene in mice: The results showed that the water-soluble extract of SMRR can significantly inhibit ear swelling caused by xylene in high dose(10 g/kg)and medium dose(5 g/kg),and the difference between the two groups was very significant(P<0.01),indicating that the water-soluble extract of SMRR has a significant difference(P<0.01).It has significant anti-inflammatory effect.(3)The effects of water-soluble extract and monomeric compounds of SMRR on phagocytosis of peritoneal macrophages in mice: The water-soluble extract of SMRR,caffeic acid,protocatechualdenyde,salvianolic acid B and rosmarinic acid were at concentrations of 2.5,2.5,2.5,0.5,0.5 μg/mL and above,respectively,compared with the cell control group,the level of phagocytic neutral red activity in rat peritoneal macrophages was significantly or significantly increased(P<0.05 or P<0.01),indicating that the water-soluble extract,salvianolic acid B,rosmarinic acid,caffeic acid and protocatechualdenyde can promote the phagocytic ability of mouse peritoneal macrophages.(4)The effects of water-soluble extract and monomeric compounds of SMRR,on the secretion of inflammatory factors by mouse peritoneal macrophages: The water-soluble extract of SMRR significantly inhibited IL-6 secretion in LPS-stimulated mouse peritoneal macrophages at a concentration of 62.5 μg/mL(P<0.05).Salvianolic acidB,rosmarinic acid and protocatechualdenyde can significantly inhibit the secretion of TNF-α,IL-6,IL-1β(P<0.05 or P<0.01)at concentrations of 15.625,15.625,3.906 μg/mL and above,respectively.Caffeic acid at a concentration of 15.625 μg/mL and above,can significantly inhibit the secretion of TNF-α,IL-6(P<0.05 or P<0.01).Compared with other monomer compounds,the secretion of TNF-α was significantly inhibited by protocatechualdenyde at a low concentration of 3.906 μg/mL(P<0.05).Compared with other monomer compounds,the secretion of IL-6 and IL-1β was significantly inhibited by protocatechualdenyde and salvianolic acid B at a low concentration of 3.906 μg/mL(P<0.05),respectively.In conclusion that the water-soluble extracts of SMRR,salvianolic acid B,rosmarinic acid,caffeic acid and protocatechualdenyde could inhibit the secretion of inflammatory factors in mouse peritoneal macrophages to different degrees,thus exerting anti-inflammatory effects,and protocatechualdenyde and salvianolic acid B have remarkable effects.