| The Allium genus includes approximately 500 species,especially in the northern hemisphere. Commonly used allium vegetables include garlic, onion, leeks, chives, scallions, which are used all over the world in different delicacies. Some allium vegetables, which are rich resource of flavone, steroidal saponins, alkaloids, fatty acid as well as sulfur-containing compounds, have been employed for medicine in the traditional medical practice such as antimicrobial, antioxidant, antithrombotic, antitumor and cancer preventing agents. In recent years, extensive research has focused on the cardiovascular potential of allium vegetables and their constituents. Semen Allii Fistulosi is the seed of Allium fistulosum L. (Liliaceae), witch is a centuried traditional Chinese medicine initially recorded in"ShenNong's Herbal". However, there are no reports on chemical compounds and myocardial ischemia activity of it.In order to investigate in the pharmacological activity of myocardial ischemia of Semen Allii Fistulosi, several animal models were evaluated, including the dog models of acute myocardial ischemia, the rat models of myocardial ischemia reperfusion injury, the anesthetized dog model of hemodynamics and myocardial oxygen consumption. The extract was shown to have significant effect of decreasing the degrees and areas of acute myocardial ischemia, lessening the injury of myocardial cell; protecting myocardial cell against injuries when myocardial ischemia reperfusion occurs, inhibiting LDH, CK from efflux and activities. In conclusion, the extract of Semen Allii Fistulosi was further proved to be active to cure myocardial ischemia.39 compounds were obtained through column chromatography of silica gel, non-polar macroporous resin, Sephadex LH-20, ODS, pre-HPLC. On the basis of spectral analysis (UV, IR, EI-MS, FAB-MS, 1H-NMR, 13C-NMR, 2D-NMR) and physicochemical property, they were identified as azelaic acid (1), (+)-Nyasol (2), ferulic acid (3), vanillic acid (4), 3- ethyl-2-(1-phenylpropyl)pentanoic acid (5), 2-methoxyhy droquinone (6), (R)-(+)-2-hydr oxy-3-phenylpropanoic acid (7), 4-hydroxybenzoic acid (8), Methyl linoleate (9), 2-meth ox ybenzene-1,4-diol (10), 2-hexyl-6-hydroxycyclohexanone (11), (E)-5,6,9-trihydroxyoct adec-7-enoic acid (12), 6 (E)-octacosaene acid (13), (E)-hexadec-2-enoic acid (14), methyl palmitate (15), hexadecanoic acid-2-hydroxy-1,3-propanediyleste (16), (8E,10E)-7-hydrox yhexadeca-8,10-dienoic acid (17), (E)-tetradeca-3-enoic acid (18), (E)-hexadeca-2-enoic acid (19), (25R)-26-O-β-D-glucopyranosyl-5,20-diene-furostan-2-one-3β,26-diol 3-O-α-L- rhamnopyranosyl-(1→2)-[α-L-rhamnopyranosyl-(1→4)]-β-D-glucopyranoside (named: Fistulosaponin A) (20), (25R)-26-O-β-D-glucopyranosyl-5,20-diene-furostan-2α,3β,26- triol 3-O-β-D-glucopyranosyl-(1→2)-β-D-glucopyranosyl-(1→2)-β-D-glucopyranoside (named: Fistulosaponin E) (21), protogracillin (22), 26-O-β-D-glucopyranosyl-(25R)- furost-5-ene-3β,22,26-triol-3-O-α-L-rhamnopyranosyl-(1→4)-α-L-rhamnopyranosyl- (1→2)-β-D-glucopyranoside (23), 26-O-β-D-glucopyranosyl-(25R)-furost-5-ene-3β,22,26- triol-3-O-2,4-di-O-(α-L-rhamnopyranosyl)-β-D-glucopyranoside (24), 26-O-β-D-glucopy ranosyl-(25R)-furost-5-ene-3β,22α,26-triol-3-O-α-L-rhamnopyranosyl-(1→4)-α-L-rhamnopyranosyl-(1→4)-[α-L-rhamnopyranosyl-(1→2)]-β-D-glucopyranoside (25), (25R)-26-O-β-D-glucopyranosyl-5-ene-furostan-2-one-3α,22α,26-triol 3-O-α-L-rhamnopyranosyl- (1→4)-α-L-rhamnopyranosyl-(1→2)-β-D-glucopyranoside (named: Fistulosaponin C) (26), (25R)-26-O-β-D-glucopyranosyl-5-ene-furostan-2-one-3β,22α,26-triol 3-O-α-L-rhamnop yranosyl-(1→2)-[α-L-rhamnopyranosyl-(1→4)]-β-D-glucopyranoside (named: Fistulosapo nin B) (27), (25R)-26-O-β-D-glucopyranosyl-5-ene-furost-2-one-3β,22α,26 triol 3-O-β-D- glucopyranosyl-(1→2)-β-D-glucopyranosyl-(1→2)-β-D-glucopyranoside (named: Fistulo saponin D) (28), (25R)-26-O-β-D-glucopyranosyl-5-ene-furostan-2α,3β,22α,26-tetraol 3- O-β-D-glucopyranosyl-(1→2)-β-D-glucopyranosyl-(1→2)-β-D-glucopyranoside (named: Fistulosaponin F) (29), diosgenin (30), yuccagenin (31), (25R)-spirost-1,4-diene-3-one-2- ol (32), (25R)-spirost-4-ene-3-one-2-ol (33), (25R)-19-norspirosta-1,3,5 (10)-triene-4- methyl-2-ol (34),(25R)-spirost-5-ene-2α,3β,6β-triol (35), (25R)-spirost-1,4-diene-3-one- 2,6-diol (36), gitogenin (37), tigogenin (38), ruscogenin (39), All of them were isolated from Semen Allii Fistulosi for the first time, and 20, 21, 26, 27, 28, 29, 34, 36 are found as novel compounds.Ischemia injuries are mainly due to reduction in the supplement of oxygen. Hypoxia and reoxygenation are principal components of ischemia/repefusion and have distinctive effects on the tissue.In vitro cell culture of hypoxia-reoxygenation injury can simulate the in vivo ischemic injury. The isolated five compounds were then evaluated for their anti-hypoxia activity on HUVECs by H/R treatment. MTT assay showed that, compared to control, H/R treatment significantly decreased the cell viability to (56.7±4.8) % (P<0.01). Treatment of HUVECs with drugs remarkably improved cell survival against H/R-induced HUVEC (P<0.05). Allium fistulosum is a daily flavor of foodstuff widely distributed in China. In order to provide scientific basis for its identification criterion, we studied on the determination of the active saponins with orthogonal experimental design of solid-liquid proportion, saturate time and ultrasonic time of ultrasound with water. HPLC-ELSD and HPLC-MS determinations of active saponins of prevention myocardial ischemia of the seeds of Allium fistulosum were established in this study. An HPLC-ELSD method was established to determine the four furostan saponins in the water extracts of 13 samples from different areas of the seeds of Allium fistulosum The results of method validation showed that this method was simple, precise and repetitive. The overall recoveries for the four analytes were between 97.91 % and 99.77 %. However,sensitivity of this method was not so satisfactory, for the contents of the saponins were less than LOD in some samples. As a result,it is applicable for the quantification of just the abundant furostan saponins in the TCM from the seeds of Allium fistulosum and identification of its plant sources. An HPLC- MS method was established to determine the seven furostan saponins in the water extracts of 20 samples from different areas of the seeds of Allium fistulosum. The results of method validation showed that this method was rapid, sensitive, accurate and repetitive. The overall recoveries for the four analytes were between 95.61 % and 99.70 %. The sensitivity of this method was several hundredfolds higher than that of the HPLC-ELSD method established in this thesis. As a result,it is applicable for the quantification of the furostan saponins in the TCM from the seeds of Allium fistulosum and identification of its plant sources.
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