| This work was supported by National Nature Science Foundation of China—Part experiment content of "Study on dose-effect relationship and synergistic interation of nourishing and activating blood effect for the compatibility of Danggui and Honghua" (No. 81274058).This dissertation was divided into four major chapters, and the major results were summarized as follows:Chapter 1. Advances in Carthamus tinctorius researchA systematical review was given about the latest research progress of Carthamus tinctorius including geographic distribution and status quo, phytochemistry, analytics, pharmacological effects, toxic and side effects, intracorporal process and the clinical use, which provided a better understanding of medicinal uses of Carthamus tinctorius.Chapter 2. Chemical investigation of the petals of Carthamus tinctorius49 compounds were isolated from the petals of Carthamus tinctorius and identified by chromatograms (silica gel, sephadex LH-20 and pre-HPLC et al) and spectroscopies (MS, IR, UV,1D-NMR and 2D-NMR) combined with literature database, which were as follows: hydroxysafflor yellow B (1*), hydroxysafflor yellow C (2*), safflomin C (3), saffloquinoside C (4), hydroxysafflor yellow A (5), anhydrosafflor yellow B (6),6-hydroxykaempferol 3-O-β-D-glucoside (7),6-hydroxykaempferol 3,6,7-tri-O-β-D-glucoside (8),6-hydroxykaempferol 3-O-β-rutinoside-6-O-β-D-glucoside (9),6-hydroxykaempferol 6,7-di-O-β-D-glucoside (10), 6-hydroxykaempferol (11), kaempferol-3-O-β-D-rutinoside (12), kaempferol (13), kaempferol-3-O-β-D-glucosyl-(1â†'2)-β-D-glucoside (14),6-methoxykaemferol (15), quercetin (16), quercetin-3,7-di-O-β-D-glucoside (17), apigenin (18), apigenin-7-O-β-apiofuranosyl-6,8-di-C-β-glucopyranoside (19),6-hydroxyapigenin-6-O-β-D-glucoside-7-O-β-D-glucuronide (20), 5,7,4’-trihydroxy-6-methoxyflavone-3-O-β-D-rutinoside (21), rutin (22), luteolin (23), scutellarein (24), (2R)-5,4’-dihydroxyl-6,7-di-O-β-D-glucopyranosyl flavanone (25), palmitic acid (26), undecanoic acid (27), hexacosanoic acid (28), (2S)-1-O-heptatriacontanoyl glycerol (29),1-hexadecanoyl propan-2,3-diol (30),4-dimethyl heptanedioic (31), n-tetratriacont-20,23-dienoic acid (32), ursolic acid (33), vanillic acid (34), p-hydroxybenzoic acid (35), p-hydroxybenzaldehyde (36), p-hydroxyacetophenone (37), gallic acid (38), rosmarinic acid (39), protocatechuic acid (40), tetrephthalic acid mono-[2-(4-carboxy-phenoxy-carbonyl)-vinyl] ester (41), syringing (42),E-1-(4’-hydroxypheny)-but-l-en-3-one (43), esculetin (44), scopoletin (45), 2-hydroxy-N-[(3S,4R,5R)-tetrahydro-4-hydroxy-5-(4Z)-4-tetrade-cenyl-3-furanyl]-, (2S)-rel-(9CI) (46*),β-sitosterol (47), daucosterol (48), stigmasterol (49), citrostadienol (50), isolumichrome (51). Among all of these 51 compounds, six compounds were quinochalcone C-glycosides, whereas compound 1 and 2 were new compounds. Meanwhile, Compound 46 was a new ceramide. Besides, compound 14,31,32,41 were isolated from Carthamus genus for the first time, and compound 21,28,29,30,33,34,38,44,45,50 were isolated from Carthamus tinctorius for the first time.Chapter 3. Pharmacological studies of Carthamus tinctorius(1) Pharmacological activity of different polar fractions of Carthamus tinctoriusAcute blood stasis syndrome rats were induced by subcutaneous injection of adrenaline hydrochloride and ice water bath. Compare with control rats, whole blood viscosity (WBV), plasma viscosity (PV), TXB2, and FIB of model group had significantly increased, and APTT, PT, TT, ADP, and 6-K-PGF1α had significantly deceased, illustrating the model was successfully reproduced. Compare with model group, medicated groups had different degrees of improvement. Except for PV and PT, other measurement indicators of 4 fold group had significant difference (P< 0.05 or P< 0.01), and endometrium remained intact with less inflammation. Besides, DPPH radical-scavenging activity test and Fe3+reducing power test were combined to evaluate the antioxidant capacity of different polar fractions. The results showed that water fraction displayed significantly high antioxidative activities.(2) Pharmacological activity of monomeric substances of Carthamus tinctoriusIn this study, the antioxidant capacity of chemical compounds were investigated on the basis of the ability to scavenge DPPH radical, ABTS radical and reduce Fe3+. The results showed that fatty acids, sterols and some organic acids from petroleum ether fraction exhibited inactive to scavenge free radical and reduce Fe3+. Flavonoids on scavenging free radical activities were bound up with their structures. The hydrogenation of C2—C3 and glycosylation of OH group at A ring were not favorable for antioxidant activities, but the glycosylation of 3-OH at C ring could increase the antioxidant activities of flavonoids, especially forming monloglycoside. There was a positive correlation between quinochalcone C-glycosides and their hydroxyl number. Compounds contained pyrogallol (eg. compound 7,24, and 38) can stably formed five-member ring with Fe3+. Quniochalcone C-glycosides also showed strong Fe3+reducing power because of their 3-OH—4-OH and 5-OH—7-carbonyl.Chapter 4. The vivo study of anhydrosafflor yellow B (AHSYB), a major active pigment from Cathamus tinctoriusA sensitive UFLC-MS/MS method for the quantification of AHSYB, a major active water-soluble pigment from Carthamus tinctorius, in rat plasma has been developed and validated. Sample preparation was achieved by protein precipitation of plasma with four volumes of methanol. Rutin was used as the internal standard (IS). The analytes were separated using a C18 column with an 8 min gradient elution, followed by mass spectrometric detection using negative electrospray ionization (ESI) in multiple reaction monitoring (MRM) mode. The method was linear in the concentration range of 25-10000 ng·mL-1 for AHSYB. Intra-day and inter-day precision was less than 6.5%. The relative error of accuracy was within ± 9.4%. The mean recovery of AHSYB was higher than 70.9%. The proposed met·hod was successfully applied to the pharmacokinetic study after intravenous (2.5 mgkg-1) and oral (30 mg·kg-1) dosing of AHSYB in normal rats. And the pharmacokinetic properties of AHSYB in rats with acute blood stasis and the differences between normal and acute blood stasis syndrome rats were also investigated. The results showed that the compound was poorly absorbed (～0.3%) and the AUC0-t, AUCo-∞ and F were all significantly lower (P<0.05) in acute blood stasis syndrome rats, suggesting that disease condition may alter the body metabolism by enhancing metabolite enzyme activity. |
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