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OBJECTIVE To investigate the inhibitory effect of natural compound XCQ-9 of Cynanchum paniculatum on the proliferation and apoptosis of Jurkat cell line of human T-cell acute lymphoblastic leukemia and its possible mechanism. METHODS Jurkat cell was used as the leukemia cell model， and MTT assay was adopted to detect the inhibitory effects of 0 （blank control）， 2.5， 5， 10， 20 and 40 μmol/L XCQ-9 on the proliferation of Jurkat cell after treated for 24， 48， 72 h. After treated with 0 （blank control）， 2.5， 5， 10 μmol/L XCQ-9 for 24 h and 48 h， the cell cycle and apoptosis were analyzed by flow cytometry. The expressions of Caspase-9， Cleaved Caspase-9， Caspase-3， Cleaved Caspase-3， poly ADP-ribose poly-merase （PARP）， Cleaved-PARP， cyclin-dependent kinase 1 （CDK1） and Cyclin B1 were detected by Western blot after treated for 24 h. RESULTS Compared with blank control group， XCQ-9 at different concentrations could significantly decrease the survival rate of Jurkat cells （P＜0.01）， and showed a dose and time-dependent manner. After 48 h treatment of 5， 10 μmol/L XCQ-9， Jurkat cell apoptosis was induced significantly （P＜0.05 or P＜0.01）， and the cell was arrested in G2 phase （P＜0.01）. After 24 h treatment of 10 μmol/L XCQ-9， the protein expressions of CDK1 and Caspase-9 were remarkably down-regulated （P＜0.01）， while the protein expressions of Cyclin B1， Cleaved Caspase-9， Cleaved Caspase-3 and Cleaved PARP were significantly up-regulated （P＜0.05 or P＜0.01）. CONCLUSIONS XCQ-9 plays anti-tumor effect through inducing G2 phase arrest to inhibit proliferation and 5008） activating Caspase pathway to increase apoptosis.
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ABSTRACT A new C21 steroidal glycoside, paniculatumoside G, together with neocynapanogenin C isolated for the first time from the natural source and two known compounds were isolated and characterized from the roots and rhizomes of Cynanchum paniculatum (Bunge) Kitag. ex H.Hara, Apocynaceae, a commonly used Traditional Chinese Medicine. On the basis of spectroscopic analysis, including HR-ESI-MS, 1D and 2D NMR spectral data, the structure of the new C21 steroidal glycoside was elucidated as neocynapanogenin H 3-O-β-D-oleandropyranoside.
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OBJECTIVE:To determine the content of paeonol in Cynanchum paniculatum by three-dimensional fluorescence coupled with ATLD algorithm. METHODS:The trilinear model was established. The principle of least square was used for ATLD. The component value was fitted by core consistency diagnosis,and inner filter effect was corrected by mathematical correction method. Fluorescence scanning condition included excitation wavelength of 250-400 nm,emission wavelength of 410-600 nm, wavelength interval of 5 nm,slit width of 5.0/5.0 nm,scanning speed of 1200 nm/min. Determination and absorption spectrum condition included scanning wavelength of 250-600 nm,scanning speed of 600 nm/min;Al(Ⅲ)was used as the sensitizer to in-crease the fluorescence intensity of paeonol. The content of paeonol was determined by HPLC. RESULTS:The linear range of pae-onol was 0.132-1.188 μg/mL(r=0.9999). RSDs of precision,stability and reproducibility tests were less than 3.0%. The recover-ies were 100.1%-104.7%(RSD=2.39%,n=6). The analysis spectrum of paeonol was almost completely overlapped with the actu-al spectrum. The results of HPLC method are very similar to those of ATLD algorithm. CONCLUSIONS:The three-dimensional fluorescence coupled with ATLD algorithm is simple,rapid,efficient and accurate,and can be used for the qualitative and quantita-tive analysis of complex system.
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@#To explore the effects of Cynanchum paniculatum paeonol on apoptosis of articular chondrocyte and the expression of Bcl-2 and Bax in rabbits with knee osteoarthritis(OA), the rabbit model was established by anterior cruciate ligament transaction and excision of partial medial meniscus. From the first week after the OA model established, Cynanchum paniculatum paeonol, triamcinolone acetonide and 0. 9% saline were injected in the right knee with 0. 2, 0. 2 and 0. 3 mg, respectively, twice a week for 5 weeks. Articular cartilage samples were taken and sliced one week after the end of treatment. The morphological changes of cells were observed under electronic microscope; the TUNEL method was employed to observe the apoptosis of articular chondrocyte; and the immunofluorescence assay was used to detect theexpression of Bcl-2 and Bax inchondrocytes. The results showed that the apoptosis rates in Cynanchum paniculatum paeonol group and triamcinolone acetonide group were significantly lower than that of the control group(P< 0. 05, P< 0. 01). Cynanchum paniculatum paeonol and triamcinolone acetonide could increase the expression of Bcl-2 and decrease the expression of Bax, thus raising the ratio of Bcl-2/Bax and inhibiting apoptosis, and the Bcl-2/Bax ratio in Cynanchum paniculatum paeonol group was much higher than that of model group(P< 0. 01).
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Objective: To investigate the chemical consituents from the active fraction in the roots and rhizomes of Cynanchum paniculatum with reversal activity of multidrug resistance. Methods: The active fraction was evaluated for reversing activities toward three human drug-resistance cell lines to clininally common used anticancer chemotherapeutic drugs. The compounds were isolated and purified by chromatography on ODS and preparative HPLC. Their structures were elucidated on the basis of chemical and spectroscopic methods, including MS, 1D, and 2D NMR spectral techniques. Results: The active fraction exhibited the significant effects in sensitization of human drug-resistance on gastric carcinoma cell line SGC-7901/VCR and human drug-resistance on colonic carcinoma cell line HCT-8/VCR to fluorouracil (5-FU). Nine compounds were isolated and identified as hancogenin B 3-O-β-D- oleandropyranoside (1), 3β,14-dihydroxy-14β-pregn-5-en-20-one (2), neocynapanogenin F (3), glaucogenin A (4), glaucogenin C (5), neocynapanogenin F 3-O-β-D-oleandropyranoside (6), glaucogenin C 3-O-β-D-thevetoside (7), glaucogenin A 3-O-β-D- oleandropyranoside (8), and 20-hydroxypregna-4,6-dien-3-one (9). Conclusion: Compound 1 is a new steroidal saponin named paniculatumoside D. C21 Steroids isolated from the active fraction in the roots and rhizomes of C. paniculatum have the potential value as multidrug resistance reversing agents.
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Objective:To determine paeonol in Cynanchum paniculatum from different habitats by HPLC to provide better quality evaluation for Cynanchum paniculatum. Methods:A Promosil C18 column (200 mm × 4. 6 mm, 5 μm) was adopted, methanol-water (55∶45) was used as the mobile phase, the flow rate was 1. 0 ml·min-1 , the detection wavelength was at 274 nm, and the injection volume was 10μl. Results:Paeonol showed a good linearity(r=0. 999 9) within the range of 0. 049 4-0. 790 8 mg·ml-1,and the re-covery was 99. 98% with RSD of 0. 17%. The content of paeonol in Cynanchum paniculatum from different habitats showed significant difference. Conclusion:The method is simple, accurate and reproducible, which can be used to determine the content of paeonol in Cynanchum paniculatum from different habitats.