| Danshen is the dry root and rhizome of Salvia miltiorrhiza Bunge.As one of the earliest and most widely used traditional Chinese medicine,Danshen possess the functions of activating blood and dredging collaterals,removing stasis and relieving pain,cooling blood and relieving carbuncle,clearing away heart and bothering.Modern pharmacological studies have shown that Danshen can protect myocardium,anti-atherosclerosis,antibacterial and anti-inflammatory,anti-oxidation,anti-tumor,hepatoprotective,improve memory dysfunction.Clinically,it can be used to treat cardiovascular and cerebrovascular diseases such as atherosclerosis,stroke,hypertension and angina pectoris.In this research,we studied the tanshinones of the 95%ethanol extracts of S.miltiorrhiza roots.By using silica gel column chromatography,Sephadex LH-20 column chromatography,C18 reversed-phase column chromatography,preparative thin-layer chromatography and HPLC,a total of 22 compounds were isolated from the ethyl acetate extracts of S.miltiorrhiza roots,the structures were identified as:tanshinone Ⅰ(1),tanshinone ⅡA(2),cryptotanshinone(3),15,16-dihydrotanshinone I(4),hydroxytanshinone ⅡA(5),tanshindiol C(6),danshenol A(7),salviolone(8),arucadiol(9),miltirone(10),hydroxymethylene tanshinone(11),methylenedihydrotanshinquinone(12),danshenxinkun A(13),danshenxinkun B(14),ursolic acid(15),1,2-dihydrotanshinol A(16),17-hydroxycryptotanshinone(17),6,12-dihydroxyabieta-5,8,11,13-tetraen-7-one(18),tanshinol E(19),danshenspiroketallactone(20),β-sitosterol(21),daucosterol(22).Among them,compound 19 is a novel compound,compound 16 and 17 are first isolated from natural products.S.miltiorrhiza f.alba C.Y.Wu&H.W.Li is a white flowered varietas of Salvia miltiorrhiza Bunge and is mainly native to Laiwu and Zhangqiu districts,Shandong Province.The chemical constituents are roughly the same as those in S.miltiorrhiza,mainly tanshinones and phenolic acids.but S.miltiorrhiza f.alba has been shown to possess unique efficacy in treating thromboangiitis obliterans(TAO),in addition to having similar pharmacological activities to S.miltiorrhiza.Previous research of our group found that a mixture of extracted phenolic acids from the roots of S.miltiorrhiza f.alba showed significantly protective effects on TAO rats.In this study,we evaluated the effect of salvianolic acid B(Sal B),one of the main phenolic acids,on TAO rats.The 75%ethanol ultrasonic extracts of S.miltiorrhiza f.alba roots were extracted with polyamide resin,MCI resin and C18 reversed-phase silica gel column chromatography after extraction,and Sal Bwith a purity more than 96%were isolated and enriched.TAO rats were developed by injection of sodium laurate into the femoral artery.Different doses of Sal B administration group(10,20,40 mg/kg)were set up,together with ligustrazine hydrochloride(15 mg/kg)as positive control.The effects of Sal B on TAO rats were investigated by observing the changes of symptoms,pathological changes of rat femoral artery,determination of the levels of thromboxane B2(TXB2),6-keto-prostaglandin F1α(6-keto-PGF1α)and endothelin-1(ET-1)in plasma using enzyme-linked immunosorbent assay,and quantitative evaluating the levels of tumor necrosis factor-a(TNF-a)and inducible nitric oxide synthase(iNOS)expression in femoral artery using immunohistochemistry.The results showed that Sal B can significantly improve the condition of the limbs of TAO rats,reduce the grade of lesions,reduce inflammation and thrombosis,decrease the levels of TXB2,ET-1 and the ratio of TXB2/6-keto-PGF1α,increase the level of 6-keto-PGF1α in plasma,and significantly inhibit the expression of iNOS and TNF-a in arterial wall.These showed that Sal B has a good protective effect on TAO by inhibiting thrombosis and inflammatory responses.This study provides a theoretical basis for the application of the roots of S.miltiorrhiza f.alba for the treatment of TAO. |
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