| Objective:In this study,two kinds of traditional Chinese medicine,Salvia Miltiorrhiza Bge.and Lepisorus ussuriensis(Regel et Maack)Ching,were selected as the main research object,and the anti-severe acute pancreatitis activity and mechanism were studied and the anti-inflammatory activity sites were screened.Methods:1.The salvianolic acid B solution was treated with sodium hydroxide to prepare the sodium danshensu and the bisphenolic acid(SMND-309)by column chromatography technique.2.The model of severe acute pancreatitis(SAP)induced by cerulein combined with lipopolysaccharide(LPS)was established in rats to investigate the anti-severe acute pancreatitis(SAP)activity of sodium danshensu and SMND-309.3.A HPLC method using caffeic acid as internal standard was established to detect the concentration of SMND-309 in plasma and various tissues of mice.The pharmacokinetic parameters and tissue distribution after intraperitoneal injection of SMND-309 in mice were investigated.4.Mouse monocyte macrophage raw264.7,was induced by lipopolysaccharide(LPS)to establish inflammatory screening model.Different polar sites(petroleum ether fraction,ethyl acetate fraction,N-butanol fraction,water fraction)of Lepisorus ussuriensis(Regel et Maack)Ching were applied to establish the model.Griess and ELISA method was used to detect the secretion level of inflammatory factors NO and PGE2 in order to determine the anti-inflammatory activity sites.Results:1.The methanol solution of salvianolic acid B is degraded by 15%sodium hydroxide,and then separated by column chromatography(subsequently by AB-8 macroporous adsorption resin and polyamide resin),the sodium danshensu and the bisphenolic acid can be prepared simultaneously.2.In the severe acute pancreatitis rat model induced by cerulein and lipopolysaccharide,compared with the model group,the congestion phenomenon of pancreatic tissue in each group of sodium danshensu and SMND-309 was significantly improved,the edema index was significantly reduced,the expression levels of IL-1?,TNF-? in serum and the levels of serum amylase(AMS)and lipase(LPS)were significantly decreased compared with the model group,the positive expression of NF-?B/p65 decreased significantly.3.After intraperitoneal injection of SMND-309,the drug showed a single-compartment distribution in mice.The peak time(tmax)is 0.083 h,the maximum blood concentration(Cmax)is 28.667 mg.L-1,and the elimination half-life(t1/2z)is 7.974 h.The distribution order of SMND-309 in the main tissues is:kidney>plasma>heart>spleen>lung>liver.4.In the screening model of anti-inflammatory activity at cell level,it was found that both ethyl acetate fraction,N-butanol fraction and water extract fraction of Lepisorus ussuriensis(Regel et Maack)Ching could significantly inhibit the release of inflammatory factor NO and PGE2 induced by LPS in raw 264.7 cells.Conclusions:1.A method for simultaneously preparing sodium danshensu and bisphenolic acid(SMND-309)was established.2.Both sodium danshensu and SMND-309 have anti-severe acute pancreatitis activity,which may be due to the inhibition of the positive expression of NF-?B/p65 and the decrease of synthesis and release level of inflammatory factor IL-1??TNF-?and achieve the effect of anti-severe acute pancreatitis.3.SMND-309 was injected intraperitoneally and distributed rapidly in the body,with the highest concentration in the kidney.4.The ethyl acetate fraction,N-butanol fraction and water extract fraction of Lepisorus ussuriensis(Regel et Maack)Ching were the anti-inflammatory active sites,and the anti-inflammatory mechanism might be related to the influence of inflammatory factors release. |
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