Proconvulsant Effect of Papaverine on Penicillin-Induced Epileptiform Activity in Rats.

AIM: Papaverine is a vasodilator agent that is an opium alkaloid. It exhibits its effects by inhibiting the phosphodiesterase enzyme. Papaverine administration is widely used to avoid symptomatic vasospasm after subarachnoid hemorrhage. We aimed, in this research, to study the effects of papaverine on the epileptic discharges stimulated by penicillin. MATERIAL AND METHODS: Adult female Wistar rats (220±30 g) were included in this research (n=30). Rats were anesthetized with urethane (1.25 g/kg) and then the left cerebral cortex was reached by opening a burr hole with a drill. Penicillin G sodium salt (500 IU)(200 IU/1 µl) was injected into the left lateral ventricle to produce epileptiform activity. Thirty minutes before penicillin G sodium injection, papaverine was administered at doses of 5, 10, 20 or 40 mg/kg intraperitoneally. RESULTS: There was no significant difference in spike frequency between the control group and the groups given 5 mg/kg, 10 mg/ kg or 40 mg/kg papaverine, while 20 mg/kg papaverine significantly increased the spike frequency (p < 0.05). CONCLUSION: Papaverine augments the epileptiform activity produced by penicillin injection. It is important to remember that papaverine might induce convulsions in patients who have epilepsy. More research is required to understand the mechanisms of the proconvulsant influence of papaverine in epilepsy.

Papaverine, a vasodilator, blocks the pore of the HERG channel at submicromolar concentration.

Papaverine, a vasodilator used as a therapeutic agent for a range of diseases, has been reported to increase the risk of occasional serious ventricular arrhythmias. To examine the mechanism for this effect, we herein tested the effects of papaverine on human ether-a-go-go (HERG) K channels expressed in HEK293 cells and Xenopus oocytes. Our results revealed that papaverine dose-dependently decreased the tail currents of HERG channel expressed in HEK293 cells with the IC50 and the Hill coefficient of 0.58 microM and 0.58, respectively, at +20 mV and 36 degrees C. The IC50 for the papaverine-induced blockade of HERG current in Xenopus oocytes was found to decrease progressively relative to depolarization (38.8, 30.0, and 24.8 microM at -10, +20, and +40 mV, respectively). The papaverine-induced blockade of HERG current was time-dependent; the fractional current was 0.92 +/- 0.03 of the control at the beginning of the pulse, but it declined to 0.18 +/- 0.06 after 6 seconds at a test potential of 0 mV. These results collectively indicate that papaverine blocks HERG channel in a concentration-, voltage-, and time-dependent manner. Two S6 domain mutations, Y652A and F656A, partially attenuated (Y652A) or abolished (F656A) the hERG current blockade, suggesting that papaverine blocks HERG channel at the pore of the channel. This was consistent with the computational simulation that showed papaverine interacts with Tyr652 and Phe656. Therefore, ventricular arrhythmias induced by papaverine could be resulted from the blockage of the HERG channel at the cardiac myocytes.

Transplant-related bronchiolitis obliterans (BOS) demonstrates unique cytokine profiles compared to toxicant-induced BOS.

Bronchiolitis obliterans (BOS - bronchiolitis obliterans syndrome - clinical diagnosis; CBO-histopathologic diagnosis), is a chronic disease process of fibrosis and cellular deposition in airways, complicating long term survival following lung transplantation. BOS is also the result of sporadic toxicant exposure, with airway signs, symptoms, and histology indistinguishable from allograft rejection. This study establishes a transplant BOS model in MHC-mismatched rats and compares their cytokine profiles and histopathology to that of our established toxicant-induced BOS model. Both models result in lung histopathology similar to human disease. Cytokines and inflammation markers that are elevated in human transplant BOS (TGFbeta, iNOS, IFNgamma) were also elevated significantly in both models. Anti-nuclear antibody was absent from all sera in transplant or toxicant models exhibiting advanced airway pathology. The cytokine osteopontin was highly elevated in BAL early in toxicant-induced BOS, but increased late in the transplant-induced BOS model. The data show that BOS is a disease of a pathologic endpoint that is induced by different triggers and processes. The highly elevated BAL osteopontin early in the toxicant-induced BOS model suggests a need for evaluation in the diagnostic setting.

Cytotoxicity of different intracavernous vasoactive drugs on cultured endothelial cells of human corpus cavernosum penis.

OBJECTIVES: To investigate the cytotoxic effect of prostaglandin E(1) (PGE(1)), a standard combination of papaverine/phentolamine, and a triple mixture of these agents on human cavernosal endothelial cells using a cell culture model. The endothelial layer of the corpus cavernosum plays an important role in signal transduction of penile erection and is directly exposed to vasoactive agents after intracavernous injection for erectile dysfunction. METHODS: Primary endothelial cells were obtained from the corpus cavernosum of 13 potent patients undergoing penile surgery. Cultured cells were exposed for 30 minutes to physiologic dilutions of 20 microg PGE(1), 30 mg papaverine/1 mg phentolamine, or the same dosages of the triple mixture of these agents, each dissolved in 5 to 50 mL sodium chloride. Lactate dehydrogenase release as a cytotoxicity marker was measured 6 hours after drug exposure, and the total cell metabolic activity was quantified after 48 hours with a 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxy-phenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS)-based assay. Additionally, the amount of viable cells was identified with a dual fluorescent staining procedure. RESULTS: The initial release of lactate dehydrogenase was elevated up to 3.2-fold in the concentrated papaverine/phentolamine and triple mixture group compared with PGE(1) and the control. After 48 hours, the papaverine-containing formulations led to a significant dose-dependent decrease in the viable cell count and metabolic activity of the cultures that was not noticed with PGE(1). CONCLUSIONS: These in vitro data strongly suggest an unfavorable effect of vasoactive agents containing papaverine on cavernosal endothelial cells. Before fibrotic changes of the smooth muscle stroma, the functionally important endothelium of the corpus cavernosum might suffer significantly from intracavernous injection therapy. Therefore, papaverine should no longer be used for this indication.

Electrocardiographic toxicity in the guinea pig.

Abnormalities of cardiac rhythm are one of the most common clinical problems in cardiology and arise as the result of either disorders of cardiac impulse formation or conduction, or a combination of both. It has been established that some classes of drugs, such as tricyclic antidepressants (e.g., imipramine), cardiac glycosides (e.g., digoxin), and Class I or Class III antiarrhythmic drugs (e.g., quinidine or amiodarone) can produce electrocardiographic toxicity in humans. It is therefore highly advisable to assess the effect of any new compound in this respect, during the early phases of drug development. This unit presents a protocol to detect the electrocardiographic toxicity of compounds in the anesthetized guinea pig.

Short-term histopathologic effects of different intracavernosal agents on corpus cavernosum and antifibrotic activity of intracavernosal verapamil: an experimental study.

OBJECTIVES: To evaluate the short-term effects of different intracavernosal agents and to investigate the antifibrotic effect of verapamil combined with these intracavernosal agents. METHODS: Forty-five Sprague-Dawley rats weighing 400 to 500 g each (mean weight 435.27 +/- 13.65 g) were equally divided into nine groups (n = 5). Papaverine (group 1), alprostadil (group 2), sodium nitroprusside (group 3), and verapamil (group 4) were injected alone intracavernously in 0.2-mL doses. Verapamil combined with papaverine, alprostadil, and sodium nitroprusside in 0.2-mL doses (0.1 mL verapamil and 0.1 mL vasoactive agent) were injected in groups 5 through 7. Group 8 was kept as a control group without injection, and isotonic saline alone was injected in group 9 during the same period. The intracavernous injection was done twice weekly with a 4-day interval. At the end of the study, total penectomy and multiple liver biopsies were performed to evaluate the histopathologic effects of the vasoactive agents and to test the liver function. RESULTS: In all groups, the structure of the corpora cavernosa was well preserved generally and appeared similar to the control tissue. However, localized edema, fibrosis, macrophage infiltration, and polymorphonuclear leukocytes were found only at the injection site. Although these findings were not different from the findings in the saline and alprostadil groups, they were slightly more extensive in the papaverine and sodium nitroprusside alone groups and also in the vasoactive agent plus verapamil groups. Although mononuclear lymphocyte infiltration was found in the portal areas, advancing into the liver parenchyma, the liver function tests were within normal limits. CONCLUSIONS: We observed that intracavernous injection, except with nitroprusside, caused focal intracavernosal fibrosis and edema. We believe these effects might not be caused by just the drug, but also by needle trauma, since general fibrosis was not observed in the short term. However, nitroprusside has a severe fibrotic effect on cavernosal tissue in the short term. Moreover, intracavernous verapamil injection could not prevent the fibrosis in the short term.

Sauropus androgynus and papaverine do not induce bronchiolitis obliterans in Sprague-Dawley rats.

BACKGROUND: In August 1995, an outbreak of bronchiolitis obliterans was observed in Taiwan. This progressive respiratory distress disease was associated with consumption of the uncooked vegetable, Sauropus androgynus. The vegetable is reportedly high in papaverine, a chemical that affects vasodilatation. This study determined whether S androgynus and papaverine induce bronchiolitis obliterans in Sprague-Dawley (S/D) rats. METHODS: The feeding doses were derived from the mean amount (in mg/kg body weight) that humans had eaten. In part A of the experiment, 30 S/D rats were fed various concentrations of S androgynus juice for eight weeks. The rats were divided into six groups, including a normal control and study groups with one time (1X), 10 times (10X, leaf only and mixed leaf and stalk) and 30 times (30X, leaf only and mixed leaf and stalk) concentration. In part B, 33 S/D rats were fed different doses of papaverine for 4 weeks. The rats were also divided into six groups, including normal control, oral feeding subgroups (1X, 10X and 20X) and intraperitoneal injection subgroups (5X and 10X). The degree of inflammation was defined semiquantitatively in the bronchioles and pulmonary vessel adventitia as including grade 0 (no inflammation), grade 1 (minimal), grade 2 (mild), grade 3 (moderate) and grade 4 (severe). RESULTS: There was no evidence of bronchiolitis obliterans in either experiment. The histopathologic findings of the lungs revealed normal or only minimal inflammatory changes in the peribronchial and perivascular adventitia in each group of both studies. The mean degree of inflammatory changes in the study groups was no different from that of control group. CONCLUSIONS: We conclude that ingestion of high-dose S anggogynus and papaverine do not induce injuries to the airways, alveoli or pulmonary vessels in the animal model of S/D rats. The species barrier may be one of the possible reasons.

Neurotoxic effects of papaverine, tetrahydropapaverine and dimethoxyphenylethylamine on dopaminergic neurons in ventral mesencephalic-striatal co-culture.

We report neurotoxic effects of papaverine, tetrahydropapaverine, dimethoxyphenylethylamine (DMPEA), and 1-methyl-4-phenylpyridinium ion (MPP+) on dopaminergic neurons in ventral mesencephalic-striatal co-culture. These compounds have been reported as mitochondrial toxins which may be implicated in the etiology and pathogenesis of Parkinson's disease. Tyrosine hydroxylase (TH)-positive neurons were decreased in dose-dependent manner by these compounds. Papaverine and MPP+ were most toxic to TH-positive neurons among the compounds tested. The order of the toxicity on TH-positive neurons was papaverine, MPP+, tetrahydropapaverine and then DMPEA. This order of toxicity was approximately the same as that reported on the inhibitory effect of these compounds on NADH-linked mitochondrial respiration and complex I activity. These findings indicate that the presence of dimethoxy residues in the catechol ring augments toxicity to dopaminergic neurons in culture.

Intraarterial infusion of high-concentration papaverine damages cerebral arteries in rats.

PURPOSE: To determine the appropriate concentration of papaverine for therapeutic intraarterial infusion against cerebral vasospasm. METHODS: We investigated histopathologic changes in cerebral arteries and brain tissue of normal Wistar rats that had received infusions of papaverine via the carotid artery. Rats were infused with 0.20 mL papaverine (concentration, 0.4% to 4.0%) via the internal carotid artery. Injury to the vascular wall was evaluated by transmission electron microscopy; pathologic changes in cerebral tissue were studied by light microscopy. RESULTS: Neither brain necrosis nor brain edema was seen under light microscopy at any concentration. At 4.0% papaverine concentration, degeneration of endothelial cells and medial smooth muscle, including vacuole formation, was observed under electron microscopy. At 1.4% concentration, degeneration of endothelial cells was seen. Extravasation of Evans blue dye was noted when drug concentration exceeded 1.4%. At 0.8% concentration, no histopathologic change was noted. CONCLUSION: On the basis of these results, we recommend a papaverine concentration of 0.8% or less for intraarterial infusion.

Changes in glutathione and cellular energy as potential mechanisms of papaverine-induced hepatotoxicity in vitro.

The purpose of this study was to elucidate the mechanism of hepatotoxicity of papaverine hydrochloride (papaver) in vitro. To evaluate the role of metabolism in the toxicity of papaver, cells were pretreated with SKF-525A or benzyl imidazole (cytochrome P450 system inhibitors) for 24 hr at 1 x 10(-5) or 1 x 10(-4) M, respectively, or with phenobarbital sodium (cytochrome P450 system inducer) for 3 days at 2 x 10(-3) M. Cells then were exposed to concentrations of papaver ranging from 1 x 10(-5) to 1 x 10(-3) M for 4 to 24 hr. Cytotoxicity was evaluated by enzyme leakage (lactate dehydrogenase) and by energy status of the cells (ATP/ADP). The role of biological reactive intermediates in the toxicity of papaver was investigated by measuring changes in cellular reduced glutathione levels (GSH), by inhibiting GSH synthesis, and by determining the production of lipid peroxidation (LPX). Papaverine produced concentration- and time-dependent increases in enzyme leakage, with significant effects occurring by the 8-hr exposure period. Pretreatment with SKF-525A or benzyl imidazole increased enzyme leakage induced by papaver especially at a later time frame (24 hr), but pretreatment with phenobarbital delayed the onset of cytotoxicity from 8 to 12 hr. Decreases in GSH levels paralleled the time course of enzyme leakage. However, the administration of buthionine sulfoximine to cell cultures dramatically decreased the time by which papaver induced cellular injury (2 hr vs 8 hr). Changes in cellular energy status (ATP/ADP) were also detected earlier than enzyme leakage (4 hr vs 8 hr). In contrast, no significant production of lipid peroxidation was noted in papaver-treated cultures. We suggest that the mechanism of papaver-induced hepatotoxicity may be related to alterations in glutathione balance of the cells and to disruption of energy homeostasis.

Coronary vasodilator reserve. Comparison of the effects of papaverine and adenosine on coronary flow, ventricular function, and myocardial metabolism.

To evaluate coronary flow reserve during cardiac catheterization, intracoronary adenosine and papaverine have been used in the clinical setting. Although papaverine maximizes coronary blood flow, it induces several toxic side effects that reduce its desirability as a coronary dilator. This investigation was designed to compare the subselective intracoronary administration of papaverine with that of adenosine in an animal model. In dogs (n = 34), we studied the effects of each agent on hemodynamics, regional myocardial blood flow, contractility (sonomicrometric and echocardiographic), metabolism (coronary arterial and venous lactate and tissue high-energy phosphates), and electrocardiographic (ST and QT intervals) parameters. Barbiturate and morphine anesthesia/analgesia was induced, and a left thoracotomy was performed. An arterial shunt was created from the left carotid artery to the left anterior descending coronary artery. Two separate groups were studied: group 1 (n = 16) for regional myocardial blood flow and mechanical function and group 2 (n = 18) for biochemical measurements. Adenosine (67 +/- 2 micrograms/min) or papaverine (6 +/- 1 mg/min) was infused into the coronary shunt at a rate of 0.5 + 0.1 ml/min for a maximum duration of 3.5 minutes. Regional myocardial blood flows were determined at control (predrug) and maximal coronary flow using radiolabeled microspheres. All hemodynamic, wall motion, biochemical, and electrocardiographic parameters were also measured at these times. Both drugs produced comparable increases in total and regional coronary blood flows (adenosine, 1.21 +/- 0.15 to 4.83 +/- 0.36 ml/min/g; papaverine, 1.21 +/- 0.05 to 4.89 +/- 0.28 ml/min/g) upon infusion into the left anterior descending coronary artery. Papaverine produced significant (p less than 0.05) changes in subendocardial ST segment electrocardiogram (-2.5 mm), QT prolongation (8 +/- 2%), myocardial creatine phosphate (47% decrease), and coronary sinus serum lactate (277% increase) compared with control. In addition, intracoronary papaverine induced an abnormal contractile pattern. No significant changes in any of these parameters (i.e., ST segment, QT prolongation, myocardial creatine phosphate level, or lactate level) were observed with intracoronary adenosine infusions. We conclude that intracoronary adenosine is comparable to papaverine for maximizing coronary blood flow without the deleterious properties observed with intracoronary papaverine.

Cytotoxicity induced by papaverine hydrochloride in fungal cell systems.

Cunninghamella echinulata was used to assess the cytotoxicity of papaverine (papaver), at concentrations ranging from 1 x 10(-5) to 1 x 10(-3) M for 1-6 days. Leakage of lactate dehydrogenase (LDH) and alanine aminotransferase (ALT), and changes in alkaline phosphatase (AP) activity and lactate/pyruvate (L/P) ratios were used as indices of cytotoxicity. Leakage of LDH occurred at the 6th day with 1 x 10(-3) M; AP activity and L/P ratios decreased after 6 days treatment with 1 x 10(-4) M, after 1 day with 1 x 10(-3) M, and after 3 days with 1 x 10(-4) M, respectively. This study provides support for the use of fungal cell systems to evaluate the toxicity of drugs and chemicals.

Toxicity assessment of papaverine hydrochloride and papaverine-derived metabolites in primary cultures of rat hepatocytes.

The present study was undertaken to assess and compare the toxic effects of papaverine hydrochloride and its metabolites. Primary cell cultures of rat hepatocytes were treated with papaverine (papaver), 3'-O-desmethyl (3'-OH), 4'-O-desmethyl (4'-OH), and 6-O-desmethyl (6-OH) papaverine at 1 x 10(-5), 1 x 10(-4), and 1 x 10(-3) M for 4, 8, 12, and 24-h periods. Cell injury was determined by: a) cell viability using the trypan blue exclusion test; b) cytosolic enzyme leakage of lactate dehydrogenase and aspartate aminotransferase; c) morphologic alterations; and d) lactate:pyruvate (L:P) ratios. Cell cultures showed concentration- and time-dependent toxic responses. For example, a decrease in cell viability and an increase in enzyme leakage were observed after cell treatment with 1 x 10(-4) and 1 x 10(-3) M papaver for 8 h; 1 x 10(-3) M 6-OH papaverine for 8 h and 1 x 10(-4) M for 24 h; and 1 x 10(-3) M 4'-OH papaverine for 24 h (P less than 0.05). Furthermore, changes in morphology correlated to cell viability and enzyme release in those cultures treated with papaver, 4'-OH and 6-OH papaverine. Some of these changes included size deformation, cell detachment from the dishes, and cell necrosis. On the other hand, an increase in L:P ratios (P less than 0.05) was detected with papaver as early as 8 h with 1 x 10(-4) and 1 x 10(-3) M and 12 h with 1 x 10(-5) M; 6-OH showed an increase in L:P ratios at 8 h with 1 x 10(-3) M and 12 h with 1 x 10(-4) M; these changes were evident with with 4'-OH at 12 h with 1 x 10(-3) M. In contrast, cells treated with 3'-OH papaverine did not show significant damage with any time period and concentration used in this study. The results of this study indicate that papaverine-derived metabolites are less cytotoxic than its parent compound, papaver. The toxicity was ranked as follows: papaver greater than 6-OH greater than 4'-OH greater than -3'-OH.

Comparison of the anti-respiratory syncytial virus activity and toxicity of papaverine hydrochloride and pyrazofurin in vitro and in vivo.

Based on reports describing their broad antiviral activity, the toxicity and antiviral efficacy of papaverine hydrochloride and pyrazofurin against respiratory syncytial virus (RSV) infection were tested in vitro in tissue culture cells and in vivo in cotton rats. Papaverine inhibited RSV replication in vitro; however, the median minimal toxic dose-median minimal inhibitory concentration ratios (MTD50:MIC50) in vitro and in vivo for papaverine were less than 4. Further work with this compound was discontinued. In contrast, pyrazofurin inhibited RSV replication in vitro (a mean MIC50 of 0.04 microgram/ml was obtained) and in vivo (RSV pulmonary titers were significantly reduced consistently in cotton rats given daily 10 mg/kg doses compared to untreated control animals). However, some toxic effects were observed in both the in vitro and in vivo tests of this compound. The remaining potential of pyrazofurin as an anti-RSV compound is discussed.

Retinal cell death occurs in the absence of retinal disc invagination: experimental evidence in papaverine-treated chicken embryos.

In an attempt to clarify the relationship between the presence of retinal cell death and the invagination of the optic vesicle, we have tested the occurrence and cytological characteristics of the retinal necrotic areas in the embryonic chicken after the administration in ovo of papaverine. Papaverine, a Ca2+ antagonist, was found to prevent the invagination of the optic vesicle. All embryonic retinae presented two distinct necrotic areas. However, these areas of cell death appeared abnormally located in the experimental, uninvaginated retina. One area was located at the transition between the retinal disc and the ventral wall of the optic vesicle; a second area was located in the dorsal wall of the optic vesicle, close to the optic stalk. We suggest that these necrotic areas represent the normal necrotic areas, should the invagination of the retinal disc have taken place. Retinal cell death appears to be programmed; it occurs whether the retinal disc invaginates or not. Cell death appears, in this experimental model, as a natural marker giving evidence that the embryonic retinal cells move from the optic stalk into the invagination retinal disc during normal eye cup formation. In addition to the uninvaginated optic vesicle the lens placode failed to invaginate in 45% of the cases, forming a lens vesicle in 55% of the remaining cases. This suggests that the two processes of invagination are governed by a different set of factors.

Effect of papaverine treatment on replication of measles virus in human neural and nonneural cells.

The replication of measles virus in human neural and nonneural cell lines in terms of growth and cytopathic effect was affected by treatment of the cells with papaverine, which increases endogenous cyclic AMP. Suppression of virus growth was most prominent in neuroblastoma cells, followed by that in epidermoid carcinoma and glioblastoma cells, whereas the suppressive effect was relatively weak in oligodendroglioma cells. The papaverine-induced suppression of virus growth in neuroblastoma cells was studied in detail. The suppression that occurred was dependent on the dose of papaverine and was reversible. By treatment with 10 microM papaverine, virus-cell interactions were modified as follows: (i) early replication steps such as adsorption, penetration, and uncoating of the virus were not affected; (ii) synthesis of viral RNAs, including genomic RNA and mRNA, was inhibited; (iii) translation of viral proteins from mRNA was not blocked; and (iv) glycosylation and transport of viral glycoproteins to the cell membrane were not inhibited, but phosphorylation was blocked. The significance of suppressed virus replication in neural cells is discussed in relation to the persistence mechanisms of measles virus in the central nervous system.