Protective Effects Of Salvia Miltiorrhiza Polyphenols And Panax Notoginseng On Angiogenesis And Myocardial Ischemia In Rats

AIM: To investigate the different effects of salvianolic acid and notoginseng triterpenes on proliferation, angiogenesis and expression of vascular endothelial growth factor in EA-hy926 cells in vitro.And to investigate the different effects of salvianolic acid and notogiaseng triterpenes on isolated heart perfusion heart perfusion and extract on pathological changes of myocardium in rats after myocardial infarction( MI) in vitro.METHODS: EA-hy926 cells were cultured in vitro.Salvianolic acid and notoginseng triterpenes at concentrations of 0.4, 0.8 and 1.2 mg·L–1were used to culture EA-hy926 cells.EA-hy926 cells in a blank control group were grown in culture solution only. Viability of cells was assessed by CCK-8, and after treated for 12 h, capillary-like structures were examined. After 24 h culture, the expression of VEGF was detected by real-time PCR.Left ventricular anterior transmural MI model was established by permanent ligation of the left descending coronary artery(LAD) with silk ligature using male SD rats.Give different dose of salvianolic acid,notoginseng triterpenes and 0.9% saline to myocardial infarction-rats through administration by gavage.After 2 weeks, rats were anesthetized using phenobarbitone(40 mg/kg body wt ip), and hearts were rapidly excised and immediately placed in ice cold Kreb-Henseleit heparin-ized solution. Hearts were perfused. A saline-filled latex balloon connected to a pressure transducer was inserted into the left ventricle. The balloon was filled with enough saline to produce an end-diastolic pressure of 16 –18 mm Hg. Digital analysis of the pressure wave was performed and displayed by an electronic polygraph. LV functions were assessed by recording: LV developed pressure(LVDP; peak systolic minus end diastolic pressure); LV end-diastolic pressure(LVEDP); maximum rate of pressure rise(dp/dtmax); and rate pressure product(RPP=heart rate×LVDP).Than, Leftventricular anterior transmural MI model was established by permanent ligation of the left descending coronary artery(LAD) with silk ligature using male SD rats.Give salvianolic acid,notoginseng triterpenes and 0.9% saline to myocardial infarction-rats through administration by gavage.After 2 weeks, rats were anesthetized using 10% Chloral hydrate, and underwent ecocardigraphy.LV functions were assessed by recording: LVIDd(LVIDd,The left ventricular internal dimension at end diastole); LVEF(LVEF,left ventricular eject fration); FS(FS, fractional shortening).Then rats were sacrificed. The segmental heart samples were collected for hematoxylin and eosin staining,masson staining. RESULTS: Salvianolic acid at 0.4, 0.8 mg·L–1, the same as notoginseng triterpenes, increased VEGF content in EA-hy926 cells. Expression of VEGF protein in the salvianolic acid at 1.2 mg·L–1group, was up-regulated as compared with notoginseng triterpenes group(P < 0.05). On isolated heart perfusion study, Salvianolic acid and notoginseng triterpenes increased LVDP,LVEDP, dp/dtmax and RPP(P < 0.05).On ecocardigraphy and pathology study,compared with the MI group,salvianolic acid and notoginseng triterpenes increased LVEF and FS(P<0.05) and decresed LVIDd(P < 0.05). There were serious myocardial necrosis,increased fibroblasts,accompanied by inflammatory cell infiltration in the rat of MI group,and slightly fuzzy cell morphology,nuclear hypertrophy,mild proliferation of fibroblasts,inflammatory cells decreasing in the salvianolic acid and notoginseng triterpenes group,while clearer cell morphology,rarefibroblasts and inflammatory cells in the compatibility of salvianolic acid,according to the HE staining analysis.The masson staining results showed that the MI group consisted of more myocardial collagen tissue,yet each treatment group consist of mainly red myocardial tissue,intermingled collagen tissue.CONCLUSION: Salvianolic acid and notoginseng triterpenes can promote LVfunctions.This analysis also provided evidence that salvianolic acid had the better effects than notoginseng triterpenes.The mechanism is maybe that salvianolic acid and notoginseng triterpenes can promote cell proliferation, angiogenesis and expression of VEGF protein.