| Salvia miltiorrhiza Bunge(SM)has been used for various diseases,including coronary heart disease,hypertension,myocardial infarction and cerebrovascular diseases,it has remarkable effects.SM has two mainly components: the lipo-soluble(tanshinones)and water-soluble(SM phenolic acids).That contains a certain amount of glycoprotein in water-soluble components.This paper mainly studies the separated four active parts from SM(total tanshinones part,total salvianolic acid part,the total salvianolic acids and glycoprotein mixed part and glycoprotein part),for the first time comparative evaluated their cardioprotective effects.1.The extraction technology of SM active parts.Using flash extractor,total tanshinones and total salvianolic acids was determined by UV spectrophotometry.Finally determine the optimum extraction: the extraction ratio is 12 BV,3 times of extraction,the concentration of the extraction solvent is 75% ethanol.The extract,plus the amount of the distilled water dissolved made into suspension,first by petroleum ether extraction,using petroleum ether-ethyl acetate(3:1)extraction,concentration,drying to obtain total tanshinones part(DSBW-1,total tanshinone content is 50.8%);the remaining water fraction concentration,drying,adding 5 times volume of 75% ethanol extract the concentrated extract,drying,total salvianolic acids and glycoprotein mixed part(DSBW-2,the content of total salvianolic acid was40.8%,the polysaccharide content of 20%);the DSBW-2 adding amount of 60%ethanol dissolution,filtration,the filtrate is added into 95% ethanol precipitation,4?for 12 h,4000 r/min speed centrifuge for 5 minutes,the supernatant was collected and concentrated under vacuum drying,the total salvianolic acids parts(DSBW-3,the content of total salvianolic acids is 54.3%);collecting precipitation,vacuum concentrating,drying,glycoprotein part(DSBW-4,the polysaccharide content of40.8%,glycoprotein content is 6.08%).2.The separation technology research of salvianolic acids.With weak polar AB-8 macroporous resin,salvianolic acids were added to distilled water,5%,10%,20% methanol gradient elution.The final determination of Danshensu is mainly concentrated in the water area,rosmarinic acid,salvianolic acid B mainlyselect the eluent of ethyl acetate-acetone-10% formic acid(8:1:0.3)was isolated from Danshensu,rosmarinic acid and salvianolic acid B.3.The research of the active parts on rat aortic rings.Isolated rat aortic rings with norepinephrine induces vascular ring contraction,to give low dose concentration of 100 ?g/mL,middle dose concentration of 500 ?g/mL,high dose concentration of 1mg/mL SM four active parts.Through the signal acquisition system,record the percentage vasodilation.The results showed that at the concentration of 1 mg/mL,diastolic DSBW-3 was the strongest,vasodilation percentage was 55.67±3.09%;DSBW-1 strength,vasodilation percentage was 47.33±2.62%.4.SM active parts of myocardial cell protection research.To study the effects of in vitro cultured rat myocardial cells,ischemia reperfusion injury model was established.Given the concentration of 2.5 ?g/mL,5 ?g/mL,10 ?g/mL SM four active parts role in model cells,by detecting OD,lactate dehydrogenase(LDH)levels and superoxide dismutase(SOD)content,the protective effect of SM comparative evaluation of four active parts of myocardial cells.The results showed that the drug concentration is 10 g/mL,DSBW-1 on the protective effect of myocardial cells,compared with the model group.This paper through the extraction separation technology of SM active parts,for the full development and utilization of resources has laid a foundation;separation of salvianolic acids,with research on the optimization of process,it has high efficiency,saving energy and protection environmental significances;comparative evaluation four active parts of cardioprotective effects from SM,clear the active parts of SM make contribution to cardioprotection,provide reference basis for the clinical application of SM. |
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