Cynanchum Paniculatum Research Articles

2,3-dihydroxy-4-methoxyacetophenone Isolated from Cynanchum paniculatum Attenuates Scopolamine-induced Impairment of Spatial Memory in Mice.

2,3-dihydroxy-4-methoxyacetophenone Isolated from Cynanchum paniculatum Attenuates Scopolamine-induced Impairment of Spatial Memory in Mice.

Protective Activity of the Ethanol Extract of Cynanchum Paniculatum (Bunge) Kitagawa on Treating Herpes Simplex Encephalitis

To date there has been no valid treatment for herpes simplex encephalitis (HSV). This study explores the protective activity of ethanol extract of Cynanchum paniculatum (bunge) kitagawa for treatment of HSV. Cell models and animal models were established and divided into 4 groups: normal group, virus group, cynanchum paniculatum group and Dexamethasone group. Flow cytometry was employed to detect apoptosis of cell model and TUNEL assay was chosen to detect apoptosis of animal tissues. The survival time of the animal models was observed. ELISA was used to measure TNF-alpha expression and the Greiss method to measure Nitric Oxide (NO) expression in the mouse brain. As a result, it was found that extract of Cynanchum paniculatum can improve the survival rate of HSV-infected mice. The extract could prevent apoptosis in the neuron cell model and reduce apoptosis rate in brain tissue after HSV infection. With the extract intervention, TNF-alpha and NO levels in brain tissue were significantly decreased in the animal model. In conclusion, the extract of Cynanchum paniculatum can prevent HSV-inducing impairment in the cell and animal model of HSE.

Simultaneous determination of ten bioactive compaounds from the roots of Cynanchum paniculatum by using high performance liquid chromatography coupled-diode array detector

Background:Cynanchum paniculatum Kitagawa belongs to Asclepiadaceae and was used in traditional medicine to invigorate blood, alleviate edema, relieve pain, and relieve toxicity for a long time.Objective:A novel and reliable high performance liquid chromatography coupled with diode array detector method has been established for simultaneous determination of 10 bioactive compounds isolated from Cynanchum paniculatum Kitagawa, one of the herbal medicines.Materials and Methods:The chromatography analysis was performed on a SHISEIDO C18 column (S-5 μm, 4.6 mm I.D. × 250 mm) at 35°C with a gradient elution of acetonitrile and water at a flow rate of 1ml/min and UV detection at 210, 230, and 280 nm.Results:The method was validated for linearity, precision, and accuracy. All calibration curves showed good linear regression (r2 > 0.9996). Limits of detection (LOD) and limits of quantification (LOQ) fell in the ranges 0.01 - 0.28 μg/ml and 0.04 - 0.83 μg/ml, respectively. The relative standard deviation (RSD) of the intra- and inter- day test, precision test were within 1.92% and 2.43%, respectively. The mean recovery of all ranged from 92.82 to 107.96% with RSD values 0.12 - 2.18%.Conclusion:The results of validation appeared that this established method was very accurate and stabilized for the quantification of 10 bioactive compounds isolated from C. paniculatum.

Anti-proliferative activity and suppression of P-glycoprotein by (−)-antofine, a natural phenanthroindolizidine alkaloid, in paclitaxel-resistant human lung cancer cells

Multidrug resistance (MDR) is a major obstacle in effective chemotherapy for cancer patients. The expression of P-glycoprotein (P-gp) in cancer cells is highly correlated with resistance to chemotherapeutic drugs. (−)-Antofine, a phenanthroindolizidine alkaloid derived from Cynanchum paniculatum, inhibits the growth of various human cancer cells. In this study, we further explored the potential of (−)-antofine to overcome the resistance induced by anti-cancer drugs. To this end, we established the paclitaxel-resistant human lung cancer cell line A549-PA by gradually exposing A549 cells to increasing concentrations of paclitaxel. As a result, the A549-PA cells acquired resistance against paclitaxel treatment and had an IC50 that was more than 200 times that of the parental A549 cells. (−)-Antofine, however, effectively suppressed the growth of both the parental and drug-resistant cells. Additional studies revealed that the anti-proliferative activity of (−)-antofine in A549-PA cells is accompanied by a down-regulation of P-gp mRNA and protein expression. The effect of reversing the multidrug resistance of A549-PA cells via (−)-antofine treatment was demonstrated an increase in intracellular rhodamine-123 accumulation, measured using FACS analysis. These findings suggest an additional chemotherapeutic value of (−)-antofine, that is, regulation of cancer cell drug resistance, in addition to its antitumor effect.

Microwave assisted ionic liquid pretreatment of medicinal plants for fast solvent extraction of active ingredients

For increasing active ingredients accessibility to the solvents, microwave assisted ionic liquid pretreatment (MILP) is introduced to destroy cell walls of medicinal plants before solvent extraction. As a new sample pretreatment method, microwave irradiation on sample pretreatment with ionic liquids (ILs) was investigated. The conditions of MILP including microwave pretreatment time and power, ionic liquid (IL) and sample ratio and particle size were optimized and Cynanchum paniculatum and its active ingredient paeonol were chosen as the representative target analyte. Under the optimized conditions, the paeonol extraction yield by MILP was higher than that of ionic liquid aqueous solution and Soxhlet extraction. In addition, the results of bioassay test showed that the antibacterial effect of crude extract by MILP was almost the same with that of soxhlet extraction. Our results demonstrate that sample pretreatment with ILs and microwave irradiation is a potential alternative method for the pretreatment of medicinal plants before solvent extraction.

Simultaneous Determination of 10 Bioactive Compounds from the Roots of Cynanchum paniculatum by using HPLC-DAD

A novel and reliable high performance liquid chromatography coupled with diode array detector method has been established, for simultaneous determination of 10 bioactive compounds isolated from Cynanchum paniculatum Kitagawa, one of the herbal medicines. The chromatography analysis was performed on a SHISEIDO C18 column (S-5µm, 4.6mm I.D.×250mm) at 35°C with a gradient elution of acetonitrile and water at a flow rate of 1ml/min, and UV detection at 210, 230 and 280nm. The method was validated for linearity, precision and accuracy. All calibration curves showed good linear regression (r 2 >0.9996). Limits of detection (LOD) and limits of quantification (LOQ) fell in the ranges 0.01–0.28µg/ml and 0.04–0.83µg/ml, respectively. The relative standard deviation (RSD) of the intra- and inter- day test, precision test were within 1.92% and 2.43%, respectively. The mean recovery of all ranged from 92.82 to 107.96% with RSD values 0.12–2.18%. The results of validation appeared that this established method was very accurate and stabilized for the quantification of 10 bioactive compounds isolated from C. paniculatum.

Neuroprotective Activity of Compounds Isolated from Cynanchum paniculatum Kitagawa

The 10 compounds, paeonol (1), 4-acetylphenol (2), 2,5-dihydroxy-4-methoxyacetophenone (3), 4-O-methyl ether Gallacetophenone (4), acetoveratrone (5), 2,5-dimethoxyhydroquinone (6), vanillic acid (7), resacetophenone (8), m-acetylphenol (9) and 3,5-dimethoxyhydroquinone (10) were isolated and identified from the 80% methanol extract of roots of Cynanchum paniculatum Kitagawa (asclepiadaceae). The evaluation for protective effect of compounds 1-10 against glutamate-induced cytotoxicity in hippocampal HT22 cell line was executed. Among them, compound 4 and 10 were showed effective relative protection of 60.55% (at 50µM) and 55.08% (at 100µM), respectively. These results suggested that compounds 4 and 10 had neuroprotective effect on the glutamate-induced neurotoxicity in HT22 cells.

Inhibition of cell growth and potentiation of tumor necrosis factor-α (TNF-α)-induced apoptosis by a phenanthroindolizidine alkaloid antofine in human colon cancer cells

Based on the potential of natural products as a source for the development of cancer chemotherapeutic agents, this study was performed to investigate the anti-proliferative and antitumor effects of antofine, a phenanthroindolizidine alkaloid derived from Cynanchum paniculatum. Antofine showed potent anti-proliferative effects in several human cancer cells with IC 50 values in the nanomolar range. Treatment with antofine for 24 h did not result in the induction of apoptotic cell death but moderately induced cell cycle arrest at G0/G1 phase and inhibited the expression of cyclin D1, cyclin E, and CDK4. In addition, antofine inhibited the transcriptional activity of β-catenin/Tcf in human colon HCT 116 cells, and the expression level of β-catenin and cyclin D1 was also down-regulated by antofine in human colon SW480 cells. Moreover, antofine potentiated tumor necrosis factor-α (TNF-α)-induced apoptosis, which was demonstrated by the increase of Annexin V-positive cell population and of the cleavage of poly (ADP-ribose) polymerase (PARP) and caspase-8. Antofine also effectively suppressed tumor growth in the HCT 116 implanted xenograft nude mouse model. Taken together, these findings suggest that antofine might be a potential candidate for the development of cancer chemotherapeutic agents derived from natural products.

Vascular dilation by paeonol — A mechanism study

The goal of this study was to investigate the mechanism underlaying the vasodilatory effect of paeonol, a major active element from the root bark of Chinese herbs Paeonia suffruticosa Andr. and Cynanchum paniculatum (Bunge) Kitagawa. Paeonol relaxed isolated rat aorta rings by 95.6% while the 10−6M forskolin-induced vasodilatation used as 100%. The EC50 of vasodilatation by paeonol is 2.9×10−4M. Although paeonol exerted endothelium-independent relaxation, L-NAME treatment inhibited paeonol-induced vasodilation of endothelium intact rings, while indomethacin did not. Both L-NAME and ODQ did not affect paeonol relaxation in the rings without endothelium. In addition, paeonol markedly elevated NO generation in cultured endothelial cells. Pre-treatment of propranolol, glibenclamide, TEA and BaCl2 did not affect paeonol relaxation of endothelium removed rings. On the other hand, pre-treated of rings (without endothelium) with paeonol markedly blocked vasoconstriction induced by AngII, PGF2α, 5-HT, dopamine, vasopressin, endothelin-1 and PE. The paeonol incubation also significantly attenuated KCl-induced contraction which mainly depended on Ca2+ influx. In Ca2+-free medium (containing 10−4M of EGTA and 60mM of KCl), paeonol suppressed the contraction curve of CaCl2. In addition, paeonol also inhibited contraction by PE in Ca2+ free solution (containing 10−4M of EGTA) which mainly relied on intracellular Ca2+ release. Whole-cell patch-clamp experiment showed that paeonol shifted the I–V curve and the peak value of calcium currents was significantly inhibited. In conclusion, our study suggested that voltage-dependent and receptor-operated Ca2+ channel, as well as intracellular Ca2+ release were all inhibited by paeonol. An intracellular Ca2+ regulatory mechanism may be responsible to potent vasodilatory effect of paeonol.

Two New C21 Steroidal Glycosides from the Stems of Cynanchum paniculatumKitag.

Abstractmagnified imageTwo new glycosides, (3β,8β,9α,16α,17α)‐14,16β : 15,20α:18,20β‐triepoxy‐16α,17α‐dihydroxy‐14‐oxo‐13,14 : 14,15‐disecopregna‐5,13(18)‐dien‐3‐yl α‐cymaropyranosyl‐(1→4)‐α‐digitoxopyranosyl‐(1→4)‐α‐oleandropyranoside (1) and (3β,8β,9α,16α,17α)‐14,16β : 15,20α : 18,20β‐triepoxy‐16β : 17α‐dihydroxy‐14‐oxo‐13,14 : 14,15‐disecopregna‐5,13(18)‐dien‐3‐yl α‐oleandropyranosyl‐(1→4)‐α‐digitoxopyranosyl‐(1→4)‐α‐oleandropyranoside (2) were isolated from the stems of Cynanchum paniculatum (Bunge) Kitag. The structures of the new compounds were elucidated by means of spectral data, including HR‐ESI‐MS, and 1D‐, and 2D‐NMR.

Isolation and Structural Study on Carbohydrates from Cynanchum otophyllum and Cynanchum paniculatum

Three new carbohydrates were isolated from the acidic hydrolysis part of the ethyl acetate extract of Cynanchum otophyllum Schneid (Asclepiadaceae) and one new carbohydrate from the ethyl acetate extract of Cynanchum paniculatum Kitagawa. Their structures were determined as methyl 2,6-dideoxy-3-O-methyl-α-D-arabino-hexopyranosyl-(1 → 4)-2,6-deoxy-3-O-methyl-β-D-arabino-hexopyranosyl-(1 → 4)-2,6-dideoxy-3-O-methyl-α-D-arabino-hexopyranoside (1), ethyl 2,6-dideoxy-3-O-methyl-β-D-ribo-hexopyranosyl-(1 → 4)-2,6-dideoxy-3-O-methyl-α-l-lyxo-hexopyranoside (2), met hyl 2,6-dideoxy-3-O-methyl-α-l-ribo-hexopyranosyl-(1 → 4)-2,6-dideoxy-3-O-methyl-β-D-lyxo-hexopyranosyl-(1 → 4)-2,6-dideoxy-3-O-methyl-α-D-arabino-hexopyranoside (3), and 2,6-dideoxy-3-O-methyl-β-D-ribo-hexopyranosyl-(1 → 4)-2,6-dideoxy-3-O-methyl-α-d-arabino-hexopyranosyl-(1 → 4)-2,6-dideoxy-3-O-methyl-α -d-arabino-hexopyranose (4), respectively, by spectral methods.

Supercritical fluid extraction of paeonol from Cynanchum paniculatum (Bge.) Kitag. and subsequent isolation by high-speed counter-current chromatography coupled with high-performance liquid chromatography-photodiode array detector

Supercritical fluid extraction (SFE) was used to extract paeonol from Cynanchum paniculatum (Bge.) Kitag. The extraction conditions including pressure, temperature, time and sample particle size were optimized with an orthogonal test. Under the optimized conditions of 15 MPa, 55 °C, 1.5 h and a sample particle size of 40–60 mesh, the process was repeated 10 times, and the extract was then collected. The yield of the crude extract from preparative SFE was 6.02%, which contained paeonol 72.02%. Paeonol of crude extract obtained by SFE, Ultrasonic Extraction (UE), Microwave-Assisted (MA), Steam Distillation (SD) and Soxhlet Extraction (SE) were further analyzed by HPLC-PDA to compare the extraction methods, and SFE is considered as the optimum one among the five processes. The SFE extract obtained was then purified successfully by high-speed counter-current chromatography (HSCCC) with a two-phase solvent system composed of petroleum ether–ethyl acetate–methanol–water (2:6:3:4, v/v) in one-step separation. From 2.42 g of the crude extract, 1.07 g of paeonol was obtained at 99.4% purity as determined by HPLC-PDA. These results indicate that SFE and HSCCC are very powerful techniques for the extraction and purification of paeonol from C. paniculatum.

Application of liquid chromatography coupled with electrospray ionization time‐of‐flight mass spectrometry for screening and quantitative analysis of C21 steroids in the roots and rhizomes of Cynanchum paniculatum

A liquid chromatography-mass spectrometry method is presented for the quantification of C21 steroids in the roots and rhizomes of Cynanchum paniculatum. Eight C21 steroids, including five steroidal aglycones and three steroidal glycosides, were simultaneously analyzed by liquid chromatography coupled with electrospray ionization time-of-flight mass spectrometry. The extracted ion current chromatograms were extracted from the total ion current chromatogram using characteristic ions produced by target compounds for peak determination. Chromatographic separation was achieved on a C18 reversed-phase column within 60 min, using an acetonitrile/water gradient. For comparision, six C. paniculatum samples from different locations were investigated by the established method, and the results indicated that the different geographical origin significantly influenced the C21 steroid composition. The method was observed to have the necessary sensitivity, selectivity, precision, and accuracy, and to be suitable for quality control of herbal medicines and their preparations.

New C 21 steroidal glycoside from Cynanchum paniculatum

A new C 21 steroidal glycoside, neocynapanogenin F 3- O-β- d-thevetoside 1, as well as its known aglycone neocynapanogenin F 2, which have an aberrant 13, 14:14, 15-disecopregnane-type skeleton were isolated from the root of Cynanchum paniculatum. Their structures were elucidated on the basis of spectroscopic data analysis. These compounds showed certain cytotoxic activities in vitro.

Fast determination of paeonol in plasma by headspace solid-phase microextraction followed by gas chromatography–mass spectrometry

Paeonol is the active component in the traditional Chinese medicines (TCMs), such as Cynanchum paniculatum, which has been used to treat many diseases, such as eczema. In this work, a simple, rapid and sensitive method was developed for the determination of paeonol in rabbit plasma, which was based on headspace solid-phase microextraction (HS-SPME) followed by gas chromatography–mass spectrometry (GC–MS). The extraction parameters of fiber coating, sample temperature, extraction time, stirring rate and ion strength were systemically optimized; the method linearity, detection limit and precision were also investigated. It was shown that the proposed method provided a good linearity (0.02–20 μg mL −1, R 2 > 0.990), low detection limit (2.0 ng mL −1) and good precision (R.S.D. value less than 8%). Finally, GC/MS following HS-SPME was applied to fast determination of paeonol in rabbit plasma at different time point after oral demonstration of Cynanchum paniculatum essential oil. The experimental results suggest that the proposed method provided an alternative and novel approach to the pharmacokinetics study of paeonol in the TCMs.

The Anti-inflammatory and Anti-nociceptive Effects of Ethyl Acetate Fraction of Cynanchi Paniculati Radix

The anti-inflammatory and anti-nociceptive effects and sedative activities of the ethyl acetate fraction of Cynanchum paniculatum (EACP) were evaluated in mice and rats by acetic acid-induced vascular permeability, arachidonic acid-induced paw edema, cotton pellet-induced granuloma formation, formalin-induced licking time, acetic acid-induced writhing response, and pentobarbital-induced sleeping time. EACP at a dose of 40 mg/kg significantly exhibited anti-inflammatory activities on acetic acid-induced vascular permeability, arachidonic acid-induced paw edema, and the late phase of formalin-induced licking time. Moreover, it showed anti-nociceptive effects on acetic acid-induced writhing responses and significant sedative effects on pentobarbital-induced sleeping time. The results demonstrated that the anti-nociceptive effects are apparently related to the sedative effects of EACP. These results support the use of Cynanchum paniculatum in relieving inflammatory pain.

The Essential Oil of Cynanchum paniculatum (Bge.) Kitag (Asclepiadaceae)

Cynanchum paniculatum (Bge.) Kitag (Asclepiadaceae) is a medicinal plant from China. The essential oil of the leaves and roots was isolated and analyzed by GC/FID and GC/MS. Eight and nine compounds were identified in the oil from leaves and roots, respectively. The predominant component found in both oils was paeonol (78.2% and 86.3%). The most striking difference among the oil samples was the presence of 3,4-dimethoxyphenylacetone in the leaf oil. The leaf oil yield was 0.5% (w/w) and the root oil yield was 1.1% (w/w).

A pair of new C-21 steroidal glycoside epimers from the roots of Cynanchum paniculatum.

Two new C-21 steroidal glycoside epimers, paniculatumosides A and B (1 and 2), based on a 13,14:14,15-disecopregnane-type skeleton, with neocynapanogenin B (3) and neocynapanogenin C (4) as two new aglycons, were isolated from the roots of Cynanchum paniculatum. Their structures were determined by spectral data interpretation.

Cytotoxic activity and G2/M cell cycle arrest mediated by antofine, a phenanthroindolizidine alkaloid isolated from Cynanchum paniculatum.

Starting with an extract derived from the root of Cynanchum paniculatum Kitagawa (Asclepiadaceae) that was active in the process of inhibiting the growth of human cancer cells in culture, a phenanthroindolizidine alkaloid antofine was isolated and identified as an active principle (IC50 = 7.0 +/- 0.2 ng/ml for A549, human lung cancer cells; IC50 = 8.6 +/- 0.3 ng/ml for Col2, human colon cancer cells). Prompted by the high potency of cancer cell growth inhibition, additional action mechanism studies were performed with antofine. Utilizing cultured Col2 cells as a model, antofine induced arrest in the G2/M phase of the cell cycle after 48 h of incubation. With wash-out experiment, colony formation was also inhibited in a dose-dependent manner. These data suggest the potential of antofine to serve as a cancer chemotherapeutic agent by virtue of arresting the cell cycle.

A glycoside from dried roots of Cynanchum paniculatum

A new glycoside, neocynapanoside A, of disecopregnane with five-and nine-membered lactone rings was isolated from a Chinese herbal drug ‘Xu-Chang-Qing’ which is the dried roots of Cynanchum paniculatum. The structure of the aglycone was deduced to be 15,20α:18,20β-diepoxy-13,14:14,15-disecopregna-5,12-dien-14(16), 18(20β)dioic acid dilactone as found in neocynapanogenin A. The total structure of the new glycoside was established as neocynapanogenin A 3-α- l-cymaropyranosyl-(1→4)-β- d-digitoxopyranosyl-(1→4)-β- d-oleandropyranoside.