Analysis of target and mechanism of rhynchophylline in treatment of inflammatory bowel disease / 中国药理学通报

Aim To investigate the feasibility and mechanism of rhynchophylline in the treatment of in-rhynchophylline flammatory bowel disease (IBD) based on network pharmacology combined with in vivo and in vitro experiments. Methods The target of rhynchophylline-IBD intersection was obtained from the database, and GO and KEGG enrichment analysis were performed. The binding of key target proteins was screened by molecular docking. In vivo the IBD model of mice was induced by sodium dextran sulfate (DSS). After seven days of rhynchophylline intervention, the signs of mice in each group were observed and DAI scores were recorded. The levels of interleukin-1β (3 (IL-1 β), my-eloperoxidase (MPO) and other inflammatory factors in colon tissue of mice were detected by ELISA. The intestinal permeability of each group was detected. In vitro experiments were conducted to establish the inflammatory model of Caco2 cells induced by DSS, and to clarify the regulatory effect of leptosinine on key targets. Results A total of 70 rhynchophylline-IBD intersection targets were screened, and enrichment analysis showed that they were related to the inflammatory prooess, PI3K-Akt and Hippo signaling pathway s. Molecular docking results showed that was most stable in binding with JAK2 and JAK1. In vivo experiment results showed that compared with model group, body weight, colon length and weight of rhynchophylline group significantly increased (P < 0. 05). DAI score, IL-1β, MPO and other inflammatory factors in colon tissue and intestinal permeability significantly decreased (P < 0. 01). In vitro experiment results showed that compared with model group, rhynchophylline group significantly promoted the proliferation of Caco2 cells (P < 0. 05). The levels of IL-6 and NO were significantly reduced (P < 0. 05). Western blot results showed that rhynchophylline could decrease the expressions of JAK2 and JAK1 (P < 0. 05). Conclusion Rhynchophylline may play a role in the treatment of IBD by inhibiting the expression of JAK2 and JAK1 proteins and reducing inflammatory response in body.

Mechanism of rhynchophylline solid lipid nanoparticles inhibiting the proliferation of airway smooth muscle cells in asthmatic model mice / 中国药房

OBJECTIVE To study the inhibitory effect mechanism of rhynchophylline solid lipid nanoparticles （Rhy-SLN） on the proliferation of airway smooth muscle cells （ASMCs） in asthmatic model mice. METHODS Asthma model was prepared by ovalbumin+calmogastrin sensitization. The primary isolation and culture of ASMCs were performed， and morphological observation and identification were also conducted [when α -smooth muscle actin （α -SMA） appeared red and Desmin appeared green in ASMCs， indicating successful cultivation of ASMCs]. The cells were divided into blank group （ASMCs of normal mice）， model group （ASMCs of asthma model mice）， Rhy-SLN group （ASMCs of asthma model mice）， recombinant suppressors of cytokine signaling 1 （SOCS1） overexpression group （ASMCs of asthma model mice transfected with SOCS1 vector）， SOCS1-RNAi group （ASMCs of asthma model mice transfected with SOCS1-RNAi vector） and SB203580 group [p38 mitogen-activated protein kinase （p38 MAPK） inhibitor， ASMCs of asthma model mice]. The cells of each group were added into the corresponding culture medium containing drug （10 μmol/L） or not containing drug for 24 hours. MTT method was used to detect the proliferation of ASMCs in asthmatic mice； Western blot assay was used to detect the protein expressions of α-SMA， interleukin-1β （IL-1β）， SOCS1， p38 MAPK and phosphorylated p38 MAPK （p-p38 MAPK） in ASMCs. RESULTS The primary ASMCs of mice varied in shape and size， presenting irregular， spindle and triangular shapes；α-SMA appeared red and Desmin appeared green， indicating successful cultivation of ASMCs. Compared with model group， ASMCs absorbance values and protein expressions of α -SMA， p38 MAPK， and p-p38 MAPK were reduced significantly in Rhy- SLN group， SOCS1 overexpression group and SB203580 E-mail：wangmeng106@163.com group， while protein expression of SOCS1 （except for group） was increased significantly （P＜0.05）； protein expressions of IL-1β was reduced significantly in ASMCs （P＜ 0.05）. ASMCs absorbance values and protein expressions of α-SMA， SOCS1， p38 MAPK and p-p38 MAPK were increased significantly in SOCS1-RNAi group （P＜0.05）. CONCLUSIONS Rhy-SLN can inhibit the proliferation of ASMCs， the mechanism of which may be associated with overexpression of SOCS1 and inhibiting the protein expressions of IL-1β and p38 MAPK.

Activation of PXR causes drug interactions with Paxlovid in transgenic mice

Paxlovid is a nirmatrelvir (NMV) and ritonavir (RTV) co-packaged medication used for the treatment of coronavirus disease 2019 (COVID-19). The active component of Paxlovid is NMV and RTV is a pharmacokinetic booster. Our work aimed to investigate the drug/herb-drug interactions associated with Paxlovid and provide mechanism-based guidance for the clinical use of Paxlovid. By using recombinant human cytochrome P450s (CYPs), we confirmed that CYP3A4 and 3A5 are the major enzymes responsible for NMV metabolism. The role of CYP3A in Paxlovid metabolism were further verified in Cyp3a-null mice, which showed that the deficiency of CYP3A significantly suppressed the metabolism of NMV and RTV. Pregnane X receptor (PXR) is a ligand-dependent transcription factor that upregulates CYP3A4/5 expression. We next explored the impact of drug- and herb-mediated PXR activation on Paxlovid metabolism in a transgenic mouse model expressing human PXR and CYP3A4/5. We found that PXR activation increased CYP3A4/5 expression, accelerated NMV metabolism, and reduced the systemic exposure of NMV. In summary, our work demonstrated that PXR activation can cause drug interactions with Paxlovid, suggesting that PXR-activating drugs and herbs should be used cautiously in COVID-19 patients receiving Paxlovid.

Content determination of four kinds of alkaloids in Uncaria by HPLC-MS-AIS / 四川精神卫生

ObjectiveTo determine the contents of four kinds of indole alkaloids （rhynchophylline， isorhynchophylline， corynoxeine， isocorynoxeine） in Uncaria by high-performance liquid chromatography-mass spectrometry-automatic internal standard （HPLC-MS-AIS）. MethodsChromatographic separation was performed using C18 column （3.0 mm×50 mm， 3.3 μm）， and the mobile phase， comprising 0.1% formic acid aqueous solution-acetonitrile （82∶18， V/V）， was eluted at a flow rate of 0.5 mL/min and column temperature of 30℃. Mass spectrometric detection was performed using an electrospray ionization source and positive multiple-reaction monitoring mode at a voltage capillary of 4 000 V. The mass-to-charge ratio （m/z） transition was 385.25/160.10 for rhynchophylline， 385.30/160.10 for isorhynchophylline， 383.25/160.15 for corynoxeine and 383.25/160.15 for isocorynoxeine， respectively. The injection volume was kept constant at 2 μL. ResultsThe linear concentration ranges of rhynchophylline， isorhynchophylline， corynoxeine and isocorynoxeine were 2.30~600.00 ng/mL （r=0.999 3）， 2.30~600.00 ng/mL （r=0.999 2）， 2.47~650.00 ng/mL （r=0.999 4） and 2.47~650.00 ng/mL （r=0.999 2）， respectively. The relative standard deviation （RSD） of precision and stability were all lower than 5.00%， the accuracy ranged from 92.40% to 104.10%， and the average recovery was 95.90%~104.60%. ConclusionHPLC-MS-AIS method is simple and accurate for the determination of four kinds of alkaloids in Uncaria， and can be used as a new method for quality control of Uncaria.

Development of a 2D-HPLC for simultaneous determination of rhynchophylline， isorhynchophylline， corynoxeine and isocorynoxeine / 四川精神卫生

ObjectiveTo develop a two-dimensional high-performance liquid chromatography （2D-HPLC） for simultaneous determination of the contents of four kinds of Uncaria alkaloids： rhynchophylline， isorhynchophylline， corynoxeine and isocorynoxeine. MethodsThe 2D-HPLC apparatus was comprised of a first chromatographic column in version Aston SC2 （3.5 mm×25 mm， 5 μm）， an intermediate column in version Aston SH C18 （3.5 mm×10 mm， 5 μm）， and an analytical column in version Aston SCB （4.6 mm×125 mm， 5 μm）. The mobile phase of the first and second liquid chromatography system were CAA-1 and mixed mobile phase （V BPI-1 basic mobile phase ∶ V MPI-1 mobile phase ∶ V OPI-1 organic mobile phase = 45∶14∶41）. The chromatographic parameters included a flow rate of 1.0 mL/min， a column temperature of 40℃， a wavelength of 254 nm， an injection volume of 500 μL and a detection time of 9.5 min. ResultsThe linear ranges of rhynchophylline， isorhynchophylline， corynoxeine and isocorynoxeine were 9.77~10 000.00 ng/mL （r=0.999 6）， 10.74~11 000.00 ng/mL （r=0.999 7）， 10.74~11 000.00 ng/mL （r=0.999 7）， 10.74~11 000.00 ng/mL（r=0.999 6）， respectively. The relative standard deviation （RSD） of precision， stability and repeatability were all less than 5.00%. The accuracy was 95.20%~104.01%， and the recovery rate was 93.63%~101.38%. ConclusionThe 2D-HPLC developed for simultaneous determination of four kinds of alkaloids in Uncaria is simple and accurate， which can be used as a new method for quality control of Uncaria.

Effects of three kinds of active ingredients of traditional Chinese medicine on dre-miR-723-5p expression in morphine-induced zebrafish brain / 中国药理学通报

Aim To study the effect of three kinds of active ingredients of traditional Chinese medicine (sinomenine, rhynchophylline and isorhynchophylline) on dre-miR-723-5p expression in morphine-induced zebrafish brain. Methods Morphine was injected intraperitoneally to zebrafish, conditional position preference (CPP) was trained and then the behavioral of animals were observed; the miRNA expression profiles of morphine-additive zebrafish were determined by small RNA sequencing; qRT-PCR was used to verify the expression of dre-miR-723-5p, three target gene databases (miRanda, miRDB, andRNAhybrid) were used to predict the target genes of dre-miR-723-5p; Kobas 3.0 was used to perform Gene Ontology (GO) and KEGG pathway analysis of these target genes. Results Morphine-induced CPP model was established successfully. Compared with control group, the resident time and movement map in drug-pair box of zebrafish in model group significantly increased. After drug administration, the resident time and movement map in drug-pair box of zebrafish decreased. The verification results of qRT-PCR were consistent with the results of small RNA sequencing. Ninety-nine putative target genes of dremiR-723-5p that were common to all three target gene databases, which were mainly enriched in biological process, cell composition and molecular function, involved in the positive regulation of MAPK signaling pathway, lysosome, cytokine-cytokine receptor interaction, and apoptosis. Conclusion Morphine can increase the expression of dre-miR-723-5p in the zebrafish brain, which can be reversed by sinomenine, isorhynchophylline, and rhynchophylline treatment, and dre-miR-723-5p may participate in the mechanism underlying morphine-induced damage of brain.

Separation and Determination of Rhynchophylline and Isorhynchophylline in Uncaria rhynchophylla by HPLC with Ionic Liquid as Mobile Phase Additives / 中国药房

OBJECTIVE： To establish a method to determine the contents of rhynchophylline and isorhynchophylline in Uncaria rhynchophylla. METHODS： The separation degree of ionic liquid 1-butyl-3-methylimidazolium chloride （C4mimCl） as mobile phase additive was compared with that of mobile phase without additives and with traditional additive triethylamine （which damaged the chromatographic column）. The optimum concentration of C4mimCl was screened and the contents of rhynchophylline and isorhynchophylline in U. rhynchophylla from 4 habitats in Jiangxi province were determined by the newly established method. The determination was performed on Dikmatech Diamonsil Plus C18 column， the mobil phase was acetonitrile-buffer （0.1％ phosphoric acid+3.0 mmol/L C4mimCl）， gradient elution. UV detection wavelength was set at 245 nm and the flow rate was 1    mL/min. Sample size was 10 μL. RESULTS： When mobile phase had no additives or 3.0 mmol/L triethylamine and 3.0 mmol/L C4mimCl were added as additives， the separation of rhynchophylline from the front peak was 1.02， 1.23 and 1.72， and the separation from the back peak was 1.06， 6.00 and 4.25， respectively. The symmetry factors were 0.81， 0.86 and 1.13， respectively. The separation of isorhynchophylline from the front peak was 0.96， 3.89 and 4.05， and the separation from the back peak was 1.02， 2.34 and 2.36， respectively. The symmetry factors were 0.88， 0.81 and 0.96， respectively. The linear range of rhynchophylline and isorhynchophylline were 4.93-157.76 （r＝0.999 9） and 4.98-159.50 μg/mL （r＝1.000）， respectively. The quantitative limits were 0.486 4， 0.793 6 μg/mL， respectively. RSDs of precision， repeatability， stability and durability tests were all less than 5％ （n＝6）. The recovery rates were 102.9％-107.8％ （RSD＝1.7％，n＝6） and 95.4％-106.3％ （RSD＝3.9％，n＝6）， respectively. The content of rhynchophylline and isorhynchophylline in U. rhynchophylla from 4 habitats were 0.758-1.343         and 1.511-1.823 mg/g， respectively. CONCLUSIONS： Addition of C4mimCl into mobile phase can enhance its separation. Established HPLC method is rapid， accurate and reproducible， which can be used for content determination of rhynchophylline and isorhynchophylline in U. rhynchophylla.

Identification of components and metabolites of Zhidong Granule in rats plasma and urine by UPLC-Q-TOF-MS / 中草药

Objective: To investigate the prototype components and metabolites of Zhidong Granule in rats plasma and urine by UPLC-Q-TOF-MS. Methods: The analysis was carried out on a Waters UPLC HSS T3 (2.1 mm × 150 mm, 1.8 μm) column, with the mobile phase of 0.1% formic acid solution (A)-acetonitrile (B) at a flowing rate of 0.3 mL/min, the column temperature was 35 ℃. Electrospray ionization (ESI) source was applied and operated in both positive and negative ion mode. Results: A total of 45 components, including 28 prototype components and 17 metabolites, were identified by comparing their retention time, ion fragmentation information with that of the blank biological samples, herb extract, and reference compounds. Conclusion: This experimental method is simple and reliable, which can provide theoretical basis for elucidating the bioactive components of Zhidong Granule.

Rhynchophylline reverses methamphetamine-induced CPP by regulating GluRl expression in zebrafish / 中国药理学通报

Aim To investigate the effect of rhyncho-phylline on methamphetamine ( METH )-induced condition place preference (CPP) zebrafish and the relationship with GluRl. Methods METH was injected intraperitoneally to zebrafish, and the expression of GluRl in zebrafish brain was detected by immunohisto-chemistry and Western blot. Results METH-induced CPP model was established after the administration of METH (40 mg • kg-1) in zebrafish. Compared to control group, the resident time in white box of model group zebrafish significantly increased; compared to model group, the resident time in white box of ket-amine (150 mg • kg-1) group and rhynchophylline (100 mg • kg-1) group dramatically decreased. Compared to control group, the number of GluRl positive cells and the GluRl expression of model group ze-brafish markedly increased; compared to model group, the number of GluRl positive cells and the GluRl expression of rhynchophylline group zebrafish evidently decreased. Conclusions GluRl is important for METH-induced CPP formation in zebrafish. Further, rhynchophylline reduces GluRl expression in METH-treated zebrafish brain, and its mechanism may be related to GluRl expression in the brain of zebrafish.

Research progresses in pharmacological effects of rhynchophylline and its isomers on nervous system / 药学学报

Uncaria rhynchophylla is one of the frequently used herbs in China, it is mainly used for heat-clearance, suppression of hyperactive liver, calming endogenous wind and arresting convulsion in traditional Chinese medicine (TCM). Alkaloids are the main active materials in Uncaria rhynchophylla, pharmacological studies have shown that Uncaria rhynchophylla and its alkaloids have comprehensive biological effects on the nervous system. Rhynchophylline is one of the most abundant alkaloids in Uncaria rhynchophylla. The recent studies demonstrate that rhynchophylline and its isomers (isorhynchophylline, corynoxine, corynoxine B) may be good drug candidates for treatment of Alzheimer's disease, Parkinson's disease, epilepsy, etc. Although the structures of the 4 alkaloids are very similar, they have different effects on nervous system. For example, corynoxine and corynoxine B exhibit better sedative effects than isorhynchophylline. Rhynchophylline and isorhynchophylline have been extensively studied. For development and utilization of rhynchophylline for nervous system disease, more studies are needed to unveil the structure-function relationship and the underlying mechanisms. Here, we summarizes the progresses the effects of rhynchophylline and its isomers on the nervous system.

Effect of rhynchophylline on expression of TH in methamphetamine-induced CPP mice / 中国药理学通报

Aim To explore the effect of rhynchophyl-line on the expression of tyrosine hydroxylase ( TH) in hippocampus of methamphetamine-induced condition place preference ( CPP) mice. Methods Metham- phetamine was injected intraperitoneally to mice, and the expression of TH was observed by immunohisto-chemistry and Western blot. Results The CPP mouse model was established successfully by methamphet-amine ( 4 mg·kg-1) . Ketamine ( 15 mg·kg-1) , rhynchophylline low dosage group (40 mg·kg-1) and rhynchophylline high dosage group ( 80 mg·kg-1) could remove the effect of methamphetamine on CPP mice. The result of immunohistochemistry showed that methamphetamine ( 4 mg·kg-1) could increase the number of TH positive cells in hippocampus while ket-amine (4 mg·kg-1), rhynchophylline (40, 80 mg· kg-1) group could attenuate the change. Western blot-ting indicated the expression of TH of model group in-creased significantly, whereas ketamine ( 15 mg· kg-1) , rhynchophylline ( 40, 80 mg·kg-1) group presented less expression. Conclusions The CPP in-duced by methamphetamine in mice may be inhibited to some extent by rhynchophylline, and its mechanism may be associated with the expression of TH.

Inhibitory effects of solid lipid nanoparticles of rhynchophylline on proliferation of rat vasculars mooth muscle cells induced by TGF-β1 / 国际中医中药杂志

Objective To observe the effect of Rhy-SLN on the proliferation of rat vascular smooth muscle cells (VSMC) induced by TGF-β1, and explore the mechanism. Methods The primary culture of rat thoracic aortic vascular smooth muscle cells was studied by tissue block culture method. The cells were divided into the control group, TGF-β1 group, TGF-β1+ the high, medium and low dosage groups of Rhy-SLN. In addition to the control group, the cells of the other groups were involved in the intervention of TGF-β1 of 20 g/L, and the high, medium and low dosage groups of Rhy-SLN cells were involved in the intervention of 25, 50, 100 mg/L of the hook teng solid lipid nanoparticles. After 24 hours of culture, MTT assay was used to detect cell proliferation inhibition rate in each group, and the cell cycle was detected by flow cytometry. The expression of c-myc and c-Fos protein in each group was detected by Western blot method. Results Compared with the TGF-β1 group, the absorbance value (0.457 ± 0.046 vs. 0.975 ± 0.049) of TGF-β1+ rhy-sln high dose group significantly decreased (P<0.01); the number of S phase cells (15.87% ± 2.47%, 15.23% ± 1.69%, 17.02% ± 2.87% vs.38.58% ± 2.68%)of TGF-β1+rhy-sln in each dose group significantly decreased(P<0.01);The c-myc(48.65 ±3.65,50.69 ± 4.16,55.29 ± 3.67 vs.68.21 ± 3.25)and c-Fos(38.78 ± 4.25,43.56 ± 3.69,46.58 ± 3.57 vs.66.54 ± 4.09) of the TGF-β1+ rhy-sln each dose group significantly decreased (P<0.01). Conclusions The Rhy-SLN can inhibit the proliferation of VSMC in rats induced by TGF-β1.Its mechanism is related to the conversion of G0/G1 phase to the S phase and the expression of the reduction of c-myc and c-fos protein.

Effect of rhynchophylline on the mRNA expression of excitatory amino acid transporter 2 and NMDA receptor 2B after oxygen-glucose deprivation in cultured rat hippocampal astrocytes / 国际脑血管病杂志

Objective To investigate the effect of rhynchophylline on mRNA expression of excitatory amino acid transporter 2 (EAAT2 ) and N-methyl-D-aspartic acid receptor 2B (NR2B) after astrocyte oxygen-glucose deprivation. Methods The subcultured third generation astrocytes from the hippocampus were inoculated into 6-well plates, and they were divided into blank control group, hypoxia-ischemia group,low-dose rhynchophylline group (0. 02 mg/ml) and high-dose rhynchophylline group (0. 2 mg/ml) after the cells were attached to the wall and grew out protrusion. The total RNAs in each group were extracted.Real-time fluorescence quantitative polymerase chain reaction was used to detect the expression levels of EAAT2 and NR2B mRNA in astrocytes of each group. Results Compared with the blank control group, the expression levels of NR2B and EAAT2 mRNA in astrocytes of the ischemia-hypoxia group were significantly higher (all P < 0. 05 ). The expression levels of NR2B and EAAT2 mRNA in the low-dose rhynchophylline group were lower than those in ischemia-hypoxia group, but there was no significant difference. The expression levels of NR2B and EAAT2 mRNA in the high-dose rhynchophylline group were significantly lower than the ischemia-hypoxia group and the low-dose rhynchophylline group (all P < 0. 05).Conclusion The expression of EAAT2 and NR2B mRNA in astrocytes of hippocampus cultured in vitro was significantly increased after ischemia and hypoxia, and rhynchophylline intervention could significantly reduce its expression in a concentration dependent manner.

Study on rhynchophylline reducing vascular endothelial cells prethrombotic state induced by TNF-α through enhancing autophagy / 中草药

Objective: To investigate the effect of rhynchophylline on HUVECs prethrombotic state induced by TNF-α through regulating autophagy. Methods: TNF-α was used to establish the HUVECs damage model. The change of autophagy was detected by transmission electron microscope and immunofluorescence technique. Western blotting was used to detect the LC3II/LC3I ratio and the expression of Beclin-1, p62, NF-κB, vWF, and PAI-1. The content of ET-1 and PGI2 in cultural supernatant was detected by ELISA. Results: TNF-α up-regulated the expression of NF-κB, vWF, and PAI-1in HUVECs, and decreased the content of ET-1 and PGI2 in cell culture supernatant (P < 0.05). In addition, more autophagosomes were observed in HUVECs, and the LC3II/LC3I ratio and beclin-1 expression were increased, as well as the expression of p62 was decreased (P < 0.05). Rhynchophylline promoted the production of autophagosome, increased the LC3II/LC3I ratio and Beclin-1 expression, and reduced the expression of p62 (P < 0.05). Moreover, the results showed that rhynchophylline down-regulated the expression of NF-κB, vWF, and PAI-1, as well as reduced the content of ET-1 and PGI2 in cultural supernatant (P < 0.05). 3-MA, an inhibitor of autophagy, abolished the effect of rhynchophylline to a certain extent (P < 0.05). Conclusion: Rhynchophylline reduces the expression of coagulative factors induced by TNF-α through enhancing autophagy, and inhibits the prethrombotic state when vascular endothelial cells have inflammatory damage.

Prevention of abdominal adhesions in rats by rhynchophylline through inhibition of Smad singnaling pathway / 药学学报

Postoperative intra-abdominal adhesion is one of the most common complications in the postoperative period. Current remedies are very ineffective to prevent the pathological outcomes except steroid hormones. Rhynchophylline is deemed as a pharmacologically active component from traditional Oriental medicine Uncaria rhynchophylla (Miq.) Jacks. (Rubiaceae). This study was designed to investigate the preventative effect of rhynchophylline on the abdominal adhesions in rats. Rhynchophylline relieved the experimental abdominal adhesion and decreased the levels of interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in the blood serum in a dose-dependent manner. The levels of transforming growth factor-β1 (TGF-β1) and connective tissue growth factor (CTGF) were reduced significantly in the peritoneal fluid. The potential mechanism of the activity is related to inhibition of the TGF-β1/Smad signaling pathway.

Effects of Ursolic Acid and Rhynchophylline in Uncariae Ramulus Cum Uncis on Human Hepatoma HepG2 Cells / 中国中医药信息杂志

Objective To investigate the effects of ursolic acid and rhynchophylline in Uncariae Ramulus Cum Uncis on human hepatoma HepG2 cells; To discuss its antihepatoma mechanism. Methods Culture the human hepatoma HepG2 cells with 50, 25, 12.5, 6.25 μmol/L ursolic acid, rhynchophylline for 24, 48, 72 hours respectively. 1%DMSO was set as negative control group. CCK-8 based cytotoxicity assay was used to detect the growth of HepG2. Acridine orange fluorescent staining was used to observe human hepatoma HepG2 cells cultured with 50 μmol/L ursolic acid, rhynchophylline and 1%DMSO for 48 hours were observed. Results Ursolic acid significantly acted as a disincentive to the growth of HepG2 cells, which was in positive correlation with time and concentration. Inhibited effect by rhynchophylline on HepG2 cell proliferation was weaker than ursolic acid. Pathological observation showed that most of the cells in rhynchophylline group showed cell shrinkage, dense pulp coating and nuclear staining edge set, and most of the cells in the ursolic acid group showed apoptosis late changes, such as nuclear cleavage and apoptotic bodies. Conclusion Ursolic acid and rhynchophyllinein Uncariae Ramulus Cum Uncis can inhibit the proliferation of HepG2 cells and can induce apoptosis.

Effect of combination of gastrodia and uncaria on pharmacokinetics of gastrodin and rhynchophylline / 中国中药杂志

To investigate the effect of the combination of gastrodia and uncaria on the pharmacokinetics of gastrodin and rhynchophylline, and determine their pharmacokinetic parameters after administration of the combination of gastrodia and uncaria at the ratio of 12∶9. Compared with uncaria group or gastrodia group, Cmax and AUC of both gastrodin and rhynchophylline were significantly increased, and tmax was retroceded by 1.5 h for rhynchophylline and 0.25 h for gastrodin. The change of tmax resulted in a 1.25 h difference in the peak time between gastrodin and rhynchophylline , which was the same between them. Uncaria shows a more effect in suppressing hyperactive Yang, while gastrodia has a balancing effect by nourishing Yin and suppressing hyperactive Yang. As a result, gastrodia could exert the effect in nourishing Yin and suppressing effect of uncaria, which could avoid the deficiency of Yang affecting Yin due to mono-treatment of uncaria. On one hand, the enhanced AUC and Cmax of gastrodin could increase the average plasma drug concentration of gastrodin, and remedy the losing effect of uncaria at the early stage; On the other hand, the increased AUC and Cmax of rhynchophylline could make up the quick elimination of gastrodia in vivo at the late stage. Their combination could lead to an increased anti-hypertensive effect with the balance of Yin and Yang. They showed unique advantages compared with simple dosage increase of western medicines. The results were consistent with the principle of TCM treatment for the hypertension due to hyperactivity of the liver Yang. In short, this study gives a good pharmacokinetic explanation of the balance of Yin and Yang and TCM treatment for both symptoms and root cause.

Rhynchophylline, One of Major Constituents of Uncariae Ramulus et Uncus Enhances Pentobarbital-induced Sleep Behaviors and Rapid Eye Movement Sleep in Rodents

Rhynchophylline (RP) is a major tetracyclic oxindole alkaloid of Uncariae Ramulus et Uncus which has been used to treat hypertension, seizures, pain and anxiety in the oriental countries. A recent report revealed that RP attenuated ischemia-induced neuronal damage and kainite-induced convulsions in animals. This study was performed to investigate whether RP enhances pentobarbital-induced sleep behaviors and modulates sleep architecture in mice. Locomotor activity was significantly inhibited by RP at 0.25 and 0.5 mg/kg, similar to 2 mg/kg diazepam (a benzodiazepine agonist) in mice. RP shortened sleep latency and increased total sleep time in a dose-dependent manner when administrated with pentobarbital (42 mg/kg, i.p.). RP also increased the number of sleeping mice and total sleep time by concomitant administration with the sub-hypnotic dosage of pentobarbital (28mg/kg, i.p.). On the other hand, RP (0.25mg/kg, p.o.) itself significantly inhibited sleep-wake cycles, prolonged total sleep time, and rapid eye movement in rats. In addition, RP also increased chloride influx in the primary cultured hypothalamic neuronal cells. In addition, we found that glutamic acid decarboxylase (GAD(65/67)) was activated by RP. In conclusion, RP augments pentobarbital-induced sleeping behaviors, and can be a candidate for treating insomnia.

Optimizaton on recipe of rhynchophylline sustained release tablet by central composite design and response surface methodology / 国际药学研究杂志

Objective To optimize the recipe of rhynchophylline sustained release tablet in order to provide a reference for industrial production of the formulation. Method With the release degree as index, the influences of single factors on the release, such as hydroxypropyl methyl cellulose(HPMC)used as an skeleton materials, ethyl cellulose(EC)as blockers and the mass fraction of polyvinylpyrrolidone (PVP)as adhesive, were evaluated to screen the optimal preparation by central composite design and response surface methodology based on the single factor investigation. Result The optimal preparation was as follows: rhynchophylline 9.5%, HPMC: 19%, EC: 7%, PVPP:2.2%, starch:61.5% and talcum powder: 0.8%. The sustained release tablet prepared by the optimized recipe of rhynchophylline had no sudden release effect and the release degree in 12 h was more than 84%. Conclusion The optimized recipe is reasonable and meets the requirements of sustained release formulation, which means it can provide a reference for industrial production of the formulation.

Effect of isorhynchophylline on head-shakes behavior and levels of monoamine neurotransmitter in model rats with Tourette syndrome / 中华行为医学与脑科学杂志

Objective To explore the effect of rhynchophylline and isorhynchophylline on headshakes behavior and levels of monoamine neurotransmitter in model rats with Tourette syndrome.Methods 40 Wistar rats were randomly divided into DOI-induced head-shakes rats (HSR group),haloperidol group,rhynchophylline group and isorhynchophylline group with 10 in each group.The inhibitory effects of rhynchophylline and isorhynchophylline were estimated by observing the HSR behavior.Dopamine (DA) and 5-hydroxytryptamine(5-HT) in the rat striatum were detected by the enzyme-linked immunosorbent assay (ELISA) method.The 5-HT2A receptor mRNA expression in prefrontal lobe cortex of the rats was measured by real-time PCR.Results Compared with HSR group,the head shakes of the rats in haloperidol group and isorhynchophylline group were significantly decreased(P＜0.01),and no change of head-shakes number was observed in rhynchophylline group (P＞0.05).There was no significant difference of head-shakes number between the haloperidol group and isorhynchophylline group(P＞0.05).Compared with HSR group,DA levels in the rat striatum were significantly decreased in isorhynchophylline group and haloperidol group((152.35± 5.80) μ~L vs (111.19±4.30) μg/L,(152.35±5.80) μg/L vs (126.42±3.17) μg/L,P＜0.01),while DA levels in the rat striatum in rhynchophylline group were not changed ((152.35±5.80) μg/L vs (142.71±5.51) μg/L,P＞0.05).There was no significant change of 5-HT2A receptor mRNA expression in rat prefrontal lobe cortex in every group(P＞0.05).Conclusion Isorhynchophylline may have an inhibitory effect on rats with DOI-induced HSR.Isorhynchophylline may decrease the DA levels in the rat stratum with DOI-induced HSR.Rhynchophylline has no significant inhibitory effect on head-shakes behavior and DA levels in the rat stratum with DOI-induced HSR.