Chelerythrine is a kind of benzo[c] phenanthridine alkaloids, with such pharmacological activities as antitumor, antibiosis and anti-inflammation, which is widely found in plant of Fumariaceae, Papaveraceae, Ranunculaceae and Rutaceae families. This article summarizes the advances in domestic and foreign studies on pharmacological effect of chelerythrine in the recent decade, in the expectation of providing scientific basis for the in-depth studies, development and utilization of chelerythrine.
Animals ; Anti-Inflammatory Agents ; pharmacology ; Antineoplastic Agents, Phytogenic ; pharmacology ; Benzophenanthridines ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Pesticides ; pharmacology ; Plants, Medicinal ; chemistry

OBJECTIVETo identify the differentially expressed proteins in the liver of Oncomelania snails induced by Eomecon chinanthe sanguinarine.

METHODSSanguinarine was extracted and purified from the dry powder of Eomecon chinanthe. Oncomelania snails were immersed in 5 mg/L sanguinarine (50 Oncomelania snails per 500 ml) or pure water for 36 h (25°C) and the livers were isolated from live snails. Total liver proteins were extracted and separated by two-dimensional gel electrophoresis. Electrophoretogram was analyzed by Image Master 2D 5.0 software. The differentially expressed proteins between sanguinarine group and pure water group were selected and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and tandem mass spectrometry sequencing of tryptic peptides.

RESULTSIn terms of protein spots, 433 ± 14 and 385 ± 12 were observed in sanguinarine group and in water group respectively. The eleven identified differentially expressed proteins included tropomyosin, hypothetical protein XP_533132, actin 87E, keratin 6A, beta-tubulin, mitochondrial inner membrane protein isoform 4, keratin 2, allatostatin precursor, ENSANGP00000020184, actin-3 and ENSANGP00000013943. Among them, hypothetical protein XP_533132 and ENSANGP00000013943 were down-regulated and the other nine proteins were up-regulated in sanguinarine group.

CONCLUSIONSanguinarine could alter the expression of proteins in livers of Oncomelania snails.

Animals ; Benzophenanthridines ; pharmacology ; Electrophoresis, Gel, Two-Dimensional ; Isoquinolines ; pharmacology ; Liver ; drug effects ; metabolism ; Proteomics ; Snails ; drug effects ; metabolism ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

OBJECTIVETo investigate the apoptosis-inducing effect of nitidine chloride in human osteosarcoma MG-63 cells and explore its mechanism.

METHODSThe effect of nitidine chloride on the proliferation of MG-63 cells was detected by colorimetric MTT assay, and the morphological changes of cells treated with nitidine chloride were observed using fluorescence and electron microscope. Flow cytometry was performed to analyze the apoptotic rate of the cells, and the protein expression levels of caspase-3, caspase-9, Bcl-2 and Bax were detected by Western blotting.

RESULTSNitidine chloride inhibited the proliferation of MG-63 cells in a dose- and time-dependent manner. Fluorescence and electron microscopy revealed distinct apoptotic changes of the cells after nitidine chloride exposure. Flow cytometry indicated that nitidine chloride induced the apoptosis of MG-63 cells in a dose-dependent manner. Exposure to nitidine chloride, as shown by Western blotting, resulted in increased expressions of cleaved caspase-3, cleaved caspase-9 and Bax and decreased expressions of pro-caspase-3, pro-caspase-9 and Bcl-2.

CONCLUSIONNitidine chloride can inhibit the proliferation of osteosarcoma cell line MG-63 by inducing cell apoptosis, the mechanism of which might be related with the activation of the caspase-dependent pathway.

Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Benzophenanthridines ; pharmacology ; Bone Neoplasms ; pathology ; Cell Line, Tumor ; Dose-Response Relationship, Drug ; Humans ; Osteosarcoma ; pathology

We have previously shown that the vasodilator effect of protopine (Pro) on rabbit aorta is related to the elevations of cAMP and cGMP. In the present study, the vasodilator mechanisms of Pro were further explored by recording the isotonic contraction of the rat aortic strips, detecting directly the intracellular free Ca(2+) concentration ([Ca(2+)](i)) with Fura-2/AM loaded vascular smooth muscle cells (VSMCs) of rat aorta, and determining the activity of protein kinase C (PKC) in rat aortic tissue with radioactive isotope gamma-32P -ATP-catalyzing assay. By recording the aortic strips contraction induced by noradrenaline (NA) and high potassium (K(+)), Pro shifted nonparallelly the concentration-response curves of NA and high K(+) to right, in which the maximal response was depressed in the presence of Pro (30 and 100 micromol/L), and the values of pD'(2) were 3.70-/+0.25 and 3.97-/+0.15 for NA and high K(+), respectively. In the Fura-2/AM loaded VSMCs, Pro (50 and 100 micromol/L) could not produce any significant change on the resting [Ca(2+)](i), but significantly decreased the [Ca(2+)](i) elevated by NA and high K(+). Pro (30 and 100 micromol/L) had no significant effect on the activity of the cytosolic and membrane PKC in the aortic strips inpretreated by NA. However, in the aortic strips pretreated by NA, the activity of membrane PKC was significantly increased and the activity of cytosolic PKC tended to be decreased by Pro, while the activity of total PKC did not change. These results suggest that Pro seems to promote the translocation of PKC from the cytosol to the membrane in the presence of NA, its vasodilator effect may be the comprehensive result of its decreasing effect on the [Ca(2+)](i) and the increasing effect on cAMP and cGMP, as well as its influence on the PKC.
Animals ; Aorta, Thoracic ; cytology ; Benzophenanthridines ; pharmacology ; Berberine Alkaloids ; pharmacology ; Calcium ; metabolism ; Cells, Cultured ; Cyclic AMP ; metabolism ; Cyclic GMP ; metabolism ; In Vitro Techniques ; Male ; Muscle, Smooth, Vascular ; cytology ; metabolism ; Norepinephrine ; pharmacology ; Protein Kinase C ; metabolism ; Rats ; Rats, Wistar ; Vasodilator Agents ; pharmacology

OBJECTIVETo explore the mechanism governing the reversal of multidrug resistance in human breast carcinoma cells by chelerythrine.

METHODSReverse transcription-polymerase chain reaction (RT-PCR) and Western blot were used to determine the expressions of protein kinase Cα (PKCα) and multidrug resistance-related genes ABCG2, ABCC1, MDR1, and P-glycoprotein (P-gp) in MCF-7Taxol cells after treatment with chelerythrine and phorbol-12-myristate-13-acetate (PMA). Also, the antitumor effect of PMA or chelerythrine and effects of PKCα activator or inhibitor in combination with paclitaxel or adriamycin on multidrug resistance in MCF-7Taxol cells were evaluated by MTT.

RESULTSRT-PCR or Western blot showed that the expressions of MDR1 and P-gp were significantly higher in MCF-7Taxol cells exposed to PMA stimuli (both P0.05).

CONCLUSIONPKCα inhibitor chelerythrine can reverse multidrug resistance in breast carcinoma cells by inhibiting the expressions of MDR1 and P-gp expression in vitro.

ATP Binding Cassette Transporter, Sub-Family B ; ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Benzophenanthridines ; pharmacology ; Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Female ; Humans

The purpose of this study is to determine 1) whether morphine postconditiong (MPostC) can attenuate the intercellular adhesion molecules-1 (ICAM-1) expression after reoxygenation injury and 2) the subtype(s) of the opioid receptors (ORs) that are involved with MPostC. Human umbilical vein endothelial cells (HUVECs) were subjected to 6 hr anoxia followed by 12 hr reoxygenation. Three morphine concentrations (0.3, 3, 30 microM) were used to evaluate the protective effect of MPostC. We also investigated blockading the OR subtypes' effects on MPostC by using three antagonists (a micro-OR antagonist naloxone, a kappa-OR antagonist nor-binaltorphimine, and a delta-OR antagonist naltrindole) and the inhibitor of protein kinase C (PKC) chelerythrine. As results, the ICAM-1 expression was significantly reduced in the MPostC (3, 30 microM) groups compared to the control group at 1, 6, 9, and 12 hours reoxygenation time. As a consequence, neutrophil adhesion was also decreased after MPostC. These effects were abolished by coadministering chelerythrine, nor-binaltorphimine or naltrindole, but not with naloxone. In conclusion, it is assumed that MPostC could attenuate the expression of ICAM-1 on endothelial cells during reoxygenation via the kappa and delta-OR (opioid receptor)-specific pathway, and this also involves a PKC-dependent pathway.
Animals ; Benzophenanthridines/pharmacology ; Endothelial Cells/cytology/*drug effects/*metabolism ; Endothelium, Vascular/cytology ; Humans ; Intercellular Adhesion Molecule-1/genetics/*metabolism ; Morphine/*pharmacology ; Naloxone/pharmacology ; Naltrexone/analogs & derivatives/pharmacology ; Narcotic Antagonists/pharmacology ; Narcotics/*pharmacology ; Protein Isoforms/metabolism ; Protein Kinase C/antagonists & inhibitors/metabolism ; Receptors, Opioid/metabolism ; Reperfusion Injury/*metabolism ; Signal Transduction/physiology ; Umbilical Veins/cytology

BACKGROUNDNitric oxide (NO) is a biologically active molecule which has been reported to protect the heart against ischemia and reperfusion injury in different species. This study aimed to test the hypothesis that nitric oxide may induce the expression of heat shock protein 72 (HSP72) which may protect the heart against ischemia.

METHODSRabbits were given intravenous saline or S-nitroso-N-acetylpenicillamine (SNAP), a nitric oxide donor, or Zaprinast, an inhibitor of cyclic guanosine monophosphate (GMP)-phosphodiesterase, which may increase myocardial cyclic GMP content. Twenty-four hours later, the rabbits were either sampled to measure HSP72, or induced with a 30-minute coronary occlusion followed by a 120-minute reperfusion, and then the infarct size was measured. Meanwhile, chelerythrine (CHE, an inhibitor of protein kinase C) was given intravenously 5 minutes before SNAP injection and the effect on HSP72 expression and infarct size was determined.

RESULTSTwenty-four hours after pretreatment, immunoblotting showed HSP72 expression increased in the SNAP group compared with control groups, and this was blocked by CHE. Myocardial infarct size in the SNAP group was smaller than that of the control group ((32.4 +/- 5.8)% vs (51.1 +/- 4.7)%, P < 0.05). Pretreated with CHE abolished the infarct size-limiting effect of SNAP ((46.0 +/- 5.1)%). Pretreatment with Zaprinast neither induced HSP72 expression nor reduced infarct size ((55.4 +/- 5.4)%).

CONCLUSIONNO induced HSP72 expression and a delayed protection to the heart via the activities of protein kinase C by a cyclic GMP-independent pathway.

Animals ; Benzophenanthridines ; pharmacology ; Cyclic GMP ; metabolism ; HSP72 Heat-Shock Proteins ; biosynthesis ; Hemodynamics ; Male ; Myocardial Infarction ; metabolism ; physiopathology ; prevention & control ; Myocardial Ischemia ; metabolism ; physiopathology ; prevention & control ; Nitric Oxide ; metabolism ; Nitric Oxide Donors ; pharmacology ; Phosphodiesterase Inhibitors ; pharmacology ; Protein Kinase C ; metabolism ; Purinones ; pharmacology ; Rabbits ; S-Nitroso-N-Acetylpenicillamine ; pharmacology

OBJECTIVETo investigate the protective effects of protein kinase C (PKC) and mitogen-activated protein kinases (MAPKs) their and mechanisms in liver ischemic preconditioning.

METHODSIn rat models of liver ischemia-reperfusion (IR) and ischemic preconditioning (IP), the liver function was evaluated by examining serum alanine aminotransferase and aspartate aminotransferase levels, and the morphological changes of the liver cells were observed under microscope. PKC activator phorbol 12-myristate 13-acetate(PMA) and inhibitor chelerythrine(CHE), as well as MEK inhibitor PD98059, were utilized to analyze the phosphorylation of PKC and P44/42 MAPKs.

RESULTSCompared with the control rats, the liver function was best protected in rats of IP group, but not in those of IP group with PD98059 or CHE treatment. The rats in IR group showed improved liver function after PMA treatment. Similarly, the phosphorylation of PKC and P44/42 MAPKs was correlated with the liver function, and highly enhanced PKC and P44/42 MAPKs activity was observed in IP and IR+PMA groups, but decreased activity in IR and IP+CHE groups.

CONCLUSIONPhosphorylation of PKC and MAPKs plays a pivotal role in the preservation of the hepatocytes during IP.

Alanine Transaminase ; blood ; Alkaloids ; pharmacology ; Animals ; Aspartate Aminotransferases ; blood ; Benzophenanthridines ; pharmacology ; Enzyme Inhibitors ; pharmacology ; Flavonoids ; pharmacology ; Ischemic Preconditioning ; Liver ; blood supply ; enzymology ; Male ; Mitogen-Activated Protein Kinases ; antagonists & inhibitors ; metabolism ; Phosphorylation ; drug effects ; Protein Kinase C ; antagonists & inhibitors ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; blood ; physiopathology ; Tetradecanoylphorbol Acetate ; pharmacology

OBJECTIVETo determine the effect of activation of lambda-opioid receptor with U50, 488H, a selective kappa-opioid receptor agonist, on the changes in electrical coupling during prolonged ischemia and to explore the possible mechanism.

METHODSThe isolated rat heart was perfused in a Langendorff apparatus. The effect of U50, 488H on electrical coupling parameters including onset of uncoupling, plateau time, slope and fold increase in r(t) was observed in isolated perfused rat heart subjected to global no-flow ischemia. The effect of U50, 488H on connexin 43 (Cx43) expression of ventricular muscle during ischemia was determined by immunohistochemistry.

RESULTSIn the prolonged ischemia model, U50, 488H concentration dependently delayed the onset of uncoupling, increased time to plateau, and decreased the maximal rate of uncoupling during ischemia. The effect of U50, 488H on electrical uncoupling parameters during ischemia was abolished by a selective kappa-opioid receptor antagonist nor-BNI or a PKC inhibitor chelerythrine. The amount of Cx43 immunoreactive signal in ventricular muscle was greatly reduced after ischemia. U50, 488H markedly increased Cx43 expression during ischemia and its effect was also attenuated by nor-BNI or chelerythrine.

CONCLUSIONThese results demonstrated that U50, 488H delayed the onset of uncoupling and plateau time, decreased the maximal rate of uncoupling and increased Cx43 expression of ventricular muscle during ischemia, and these effects of U50, 488H were mediated by kappa-opioid receptor, in which activation of PKC was involved. The effect of U50, 488H on electrical coupling during ischemia was probably correlated with preservation of Cx43 in cardiac muscle.

3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cyclohexyl)-benzeneacetamide, (trans)-Isomer ; pharmacology ; Animals ; Benzophenanthridines ; pharmacology ; Connexin 43 ; metabolism ; Female ; Heart ; drug effects ; In Vitro Techniques ; Myocardial Ischemia ; metabolism ; physiopathology ; Myocardium ; metabolism ; Naltrexone ; analogs & derivatives ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Receptors, Opioid, kappa ; metabolism ; Signal Transduction ; drug effects

The purpose of this study is to investigate the effect of chelerythrine on the hypertrophy of cardiomyocytes of neonatal rats induced by different glucose levels and its mechanism. Using cultured neonatal ventricular myocytes as a model, groups were divided as: control (5 mmol x L(-1)); high glucose level (10, 15, 20, and 25.5 mmol x L(-1)); high glucose level (25.5 mmol x L(-1)) add different concentrations of chelerythrine (1 and 8 micromol x L(-1)); and control glucose level (5 mmol x L(-1)) add different concentrations of chelerythrine (1 and 8 micromol x L(-1)). Different groups of cardiomyocytes after adding corresponding treat factors were cultured for 48 hours. Cardiomyocytes' diameters and protein level were measured and the expression of PKC-alpha, PKC-beta2, p-PKC-alpha, and p-PKC-beta2 were measured by Western blotting. Compared with control group, neonatal myocytes cultured in high glucose levels showed increased cellular volumes, protein level and expression of PKC-alpha, PKC-beta2, p-PKC-alpha, p-PKC-beta2. When chelerythrine was added, cellular volumes, protein level and expression of PKC-alpha, PKC-beta2, p-PKC-alpha, p-PKC-beta2 were significantly reduced. But in 1 micromol x L(-1) chelerythrine group, the expression of PKC-beta2 was not significantly reduced. The result suggested that chelerythrine can reverse the hypertrophy induced by different glucose levels on the cardiac myocytes, it may have protective effect against diabetic cardiomyopathy via PKC passageway.
Animals ; Animals, Newborn ; Benzophenanthridines ; pharmacology ; Cells, Cultured ; Diabetes Mellitus, Experimental ; drug therapy ; metabolism ; Dose-Response Relationship, Drug ; Glucose ; administration & dosage ; Hypertrophy ; chemically induced ; pathology ; Hypoglycemic Agents ; pharmacology ; Myocytes, Cardiac ; drug effects ; pathology ; Phosphorylation ; Protein Kinase C ; antagonists & inhibitors ; metabolism ; Protein Kinase C beta ; Protein Kinase C-alpha ; metabolism ; Rats ; Rats, Sprague-Dawley