Studies on hydragogue drugs: light and electron microscopic examination of the isolated rat colonic mucosa exposed to deoxycholic acid and synthetic surfactants

Sacs of the stripped and everted, isolated descending rat colon were incubated for 2 hours in presence of the following surfactants at the mucosal side: Dodecylsulphate (DDS), dioctylsulphosuccinate (DOSS), cetrimonium bromide (CTMAB), Triton X100 and deoxycholic acid (DOC). After tissue fixation, the sacs were processed for light microscopy (LM) and for scanning (SEM) and transmission (TEM) electron microscopy. All three methods revealed that DOSS (1.3 X 10(-4) and 2.6 X 10(-4) mol/l, CTMAB (5 X 10(-5) and 1 X 10(-4) ) and Triton (2 X 10(-5), 5 X 10(-5) and 1 X 10(-4) ) caused only minor or moderate changes compared to parallel controls, as did also DDS at 1 X 10(-5) and 2 X 10(-5) mol/l. DDS at 2 X 10(-4) and 4 X 10(-4) mol/l and DOC at 1.5 X 10(-4) and 3 X 10(-4) mol/l caused more prominent changes. LM showed swollen, vacuolated cells with pycnotic nuclei; many of these cells seemed to be extruded. According to SEM, cells thus affected were most abundantly localized to the normal extrusion zone at the borders of the crypt-surface epithelial cell units. DOC tended to cause a more generalized affection within the units than DDS. In spite of these deleterious effects, gaps corresponding to missing epithelial cells were not observed. TEM indicated the mechanism responsible for restoration of epithelial continuity in spite of extensive cell loss: The remaining epithelial cells seemed to flatten out and re-establish cell-to-cell contact by pseudopod formation along the basement lamina. This repair mechanism seemed to operate at a rapid rate; however, incomplete closure of cellular gaps i.e. small denuded parts of the basement lamina were occasionally observed. The results of this study are discussed in relation to a functional study under identical experimental conditions (Gastroenterol. Clin. Biol. 1981, 5, 124), in which these surfactants caused a significant alteration of normal colonic transport function.     

Methoxyethylmercury nephrotoxicity: effects on enzymuria and kidney function

The fungicide, methoxyethylmercury chloride, was given in a saline solution to four groups of Sprague-Dawley C D rats (5 , 5 ) as a single injection (IP) of 0, 0.5, 1.0, and 2.0 mg Hg/kg. In a three-day period, no changes were observed in urine collected every 24 h from rats given 0 or 0.5 mg Hg/kg; 1 mg Hg/kg induced only a transient increase of urine gamma glutamyl transferase (x 4) and alkaline phosphatase (x 2.5) on the day 2; 2.0 mg Hg/kg caused an early increase of enzymuria (day 1 and day 2) and a decrease of Na⁺, Cl^–, K⁺, urea, and creatinin excretion. Urine enzymes and total mercury excretion were higher in males. These time-related variations of enzymuria, compared to previous results with Hg Cl₂, could reflect the existence of metabolites more toxic than the native compound.     

Effects of morphine on rat kidney glomerular podocytes: a scanning electron microscopic study

Degenerative kidney changes are associated with heroin use in human addicts, but it is not known whether these changes result from exposure to the opioid or from contaminants in street heroin. In the present study, 4-6-month-old rats each received 1 subcutaneous pellet containing 75 mg of morphine or placebo, followed 3 days later by implantation of 2 additional morphine or placebo pellets. Seven days after implantation of the first pellet, the rats were killed by aldehyde perfusion. The right kidney was excised, and coronal slices were prepared for scanning electron microscopy. Micrographs were taken at 5000X and were scored on the presence of short or long microprojections (a score of '1' indicating few and a score of '4' indicating many). Morphine significantly altered the frequencies of scores for long microprojections, suggesting that morphine treatment increased the number of microprojections on glomerular podocytes. No changes in filtration slits, pedicels, or blebbing (foval enlargements) were noted. The data support the view that kidney degeneration associated with opioid abuse reflects effects of opioids per se, and they are consistent with microprojection changes as a function of altered intracellular cyclic AMP levels.     

Comparative light and electron microscopic observations of the lesive effects of two non-steroid anti-inflammatory drugs plus stress on rat gastric mucosa

In this histological study it has been demonstrated that a single-dose administration of piroxicam, at the same dosage (4 mg/kg) as droxicam, has a greater erosive potential on the gastric mucosa of rats later exposed to cold stress. For this purpose the depth of all lesions found was evaluated by light microscopy and results showed that piroxicam produces lesions deeper and more numerous than those of droxicam. The transmission and scanning electron microscopic studies showed that the lesive mechanism was very similar for both drugs and that both local and general factors induced by these drugs and stress come into play. Absorption or penetration and uptake by the cells of the mucosa have been considered among the most important local factors in the development of erosive gastric lesions caused by non-steroid anti-inflammtaory drugs. As this absorption is in direct relation to the depth of the lesions, it can be considered from the results of this study that the lesser lesive effect of droxicam on the gastric mucosa when compared to that of piroxicam is due to the fact that, owing to its hydrolysis to piroxicam the absorption rate is slower.     

The effects of oral glutamine on cisplatin-induced nephrotoxicity in rats

The antioxidant activity of the amino acid glutamine was investigated to obtain protection against peroxidative damage in rat kidney and nephrotoxicity induced by the treatment with a single dose of the antitumoral cisplatin (5mgkg(-1) body weight). The animals were divided into four treatment and control groups of six rats each (n=6). Cisplatin was injected i.p. and glutamine (300mgkg(-1) body weight) was given by gavage 24h before the cisplatin injection. After 24h and 7 days of cisplatin administration, the rats were sacrificed. A single dose of cisplatin resulted in significant reduction in body weight and creatinine clearance, and higher urinary volumes were observed in all groups treated with this antitumor drug (P<0.05). Renal tissue from cisplatin-treated rats showed an increase in malondialdehyde production and increase in glutathione contents 24h and 7 days after cisplatin administration. Pretreatment of rats with glutamine substantially inhibited the increase in the levels of renal glutathione induced by cisplatin 24h after the i.p. injection. The malondialdehyde in the renal tissues was significantly reduced 7 days after cisplatin treatment. However, the reduction in the peroxidative damage did not reach the value of the control group. The protective effects obtained by glutamine pretreatment in peroxidative alterations were not observed in the other parameters studied. These results suggest that glutamine partially protect against cisplatin-induced lipid peroxidation damage, but it was not enough to inhibit cisplatin-induced nephrotoxicity in rats.     

Pharmacokinetics of codeine after parenteral and oral dosing in the rat

The pharmacokinetics of codeine was examined in six male Sprague-Dawley rats following iv bolus (3 mg/kg) and oral (5 mg/kg) codeine in a randomized crossover design. Whole blood concentrations of codeine and its O-demethylated metabolite, morphine, were determined by HPLC with electrochemical detection. Following iv codeine administration, the distribution and elimination are best described by an open two-compartment model. The weight normalized volume of distribution of (Vdarea) and total body clearance (CL) were 5.1 +/- 1.7 liters/kg and 6.2 +/- 1.5 liters/kg/hr, respectively. Mean residence time of codeine averaged 34.1 +/- 6.9 minutes. The ratio of AUCmorphine/AUCcodeine was 0.05 +/- 0.02. The absolute bioavailability of codeine calculated was 8.3 +/- 3.2%, indicating extensive first pass metabolism of codeine. An equivalent amount of codeine and morphine were present in the rat following oral codeine. Thus, the amount of morphine formed following codeine administration depends on the route of codeine administration.     

Pharmacokinetics of nitroglycerin after parenteral and oral dosing in the rat

The pharmacokinetics of nitroglycerin was characterized in detail using venous plasma after different intravenous bolus doses (0.15-2.48 mg/kg), intra-arterial infusion (8.2 micrograms/min over 5 h), and oral doses (7-100 mg/kg). Venous plasma clearance was found to be approximately 650 mL/kg and was independent of the intravenous or intra-arterial dose. This confirmed earlier reports that the venous plasma clearance of nitroglycerin in rats exceeded the value of normal cardiac output. A terminal half-life of approximately 15 min was observed after high intravenous bolus doses of nitroglycerin. This slow disappearance phase was likely rate limited by redistribution of drug back into the plasma. The bioavailability of oral nitroglycerin (F) showed an apparent Michaelis-Menten dependency on dose. F was less than 5% at doses less than 20 mg/kg, but increased to a plateau of approximately 20% from 50-100 mg/kg. First-pass metabolism of nitroglycerin is thus apparently controlled by at least two systems (sites or enzymes). Coadministration of mannitol hexanitrate, a potential competitive inhibitor of first-pass metabolism, did not increase F.     

Pharmacokinetics of pentaerythritol tetranitrate following intra-arterial and oral dosing in the rat

The pharmacokinetics of pentaerythritol tetranitrate (2,2-bis(hydroxymethyl)-1,3-propanediol tetranitrate, 1) were studied in rats following a single intra-arterial or oral dose (2 mg/kg) of the 14C-labeled drug. Blood levels of the tetranitrate and its metabolites were determined using a thin-layer radiochromatographic procedure. The apparent systemic clearance of 1 was 0.61 +/- 0.16 L/min/kg (mean +/- SD, n = 6) which exceeded the value of normal cardiac output in rats. The steady-state volume of distribution was 4.2 +/- 1.1 L/kg (n = 6), and the elimination half-life was estimated at 5.8 +/- 0.6 min (n = 6). Blood levels of 1 were only detectable (higher than 4.0 ng/mL) in three of the six rats examined after the oral dose. The trinitrate derivative (2,2-bis(hydroxymethyl)-1,3-propanediol trinitrate, 2) the active metabolite of 1, was not detectable following oral dosing with the tetranitrate. The oral bioavailability of 1 was in the range of 0-8%. In spite of the low water solubility of 1 (i.e., 1 microgram/mL), a rather high fraction of the radioactive oral dose [25.7 +/- 10.3% (n = 4) versus 62.4 +/- 14.5% (n = 4) from the intra-arterial dose] was recovered in the urine. A significant portion of the intra-arterial dose (32.7 +/- 11.0%, n = 4) was eliminated in feces, indicating enterohepatic recycling of radioactivity. Analysis of the metabolite pattern in urine indicated extensive metabolism of 1, 2, and the dinitrate derivative 3 (2,2-bis(hydroxymethyl)-1,3-propanediol dinitrate). Less than 0.2% of the dose was recovered as unchanged drug and 2 following either route of administration.     

Antiarrhythmic, electrophysiological and haemodynamic effects of prolonged oral dosing with Org 7797 in the anaesthetized rat.

The antiarrhythmic, electrophysiological and haemodynamic effects of chronic oral administration of Org 7797 ((16 alpha,17 beta)-17-methylamino-oestra-1,3,5(10)-triene-3, 16-diol-(Z)-2-butonedioate) were studied in rats. During dosing (10 mg kg-1 twice a day for 10 days) no effects on the electrocardiogram, monitored in conscious animals, were observed despite modest reductions (15-18%) in the maximum rate of depolarization of papillary muscle excised 1 or 6 h after completion of the dosing regime. Following anaesthesia, Org 7797 reduced the severity of arrhythmias induced by coronary artery occlusion and prevented the accompanying decrease in the ventricular fibrillation threshold (VFT) at 1 h after completion of dosing. By 6 h the effect on VFT had waned but protection against ischaemia-induced arrhythmias was retained despite a substantial decrease in Org 7797 plasma levels. Drug treatment did not modify arterial blood pressure, heart rate or stroke volume. We conclude that Org 7797 given chronically via the oral route exerts antiarrhythmic actions which may, at least in part, be due to sodium-channel block. In addition, our results suggest the presence of an active metabolite. The protective effects of Org 7797 were seen in the absence of electrocardiographic or haemodynamic changes suggesting that multiple oral doses of Org 7797 do not compromise normal cardiac function.     

Atypical dose-route-dependent food effects of eplerenone in the dog: presence of food effects following intravenous dosing and lack of food effects of following oral dosing

This study was conducted to investigate why a food effect was observed following an intravenous dose of eplerenone (EP) in the dog, but not following oral dosing. Three female dogs were implanted with a chronic portal vein access port and received radiolabeled EP doses orally (15 mg/kg in solution) and intravenously (7.5 mg/kg via cephalic and portal veins) under fasted and fed conditions. Mean AUC values for EP after infusion through the cephalic vein were 23.0 +/- 2.7 and 18.2 +/- 1.1 h.microg/mL under fasted and fed conditions, respectively. Corresponding values after infusion through the portal vein were 20.7 +/- 3.2 and 12.9 +/- 1.3 h.microg/mL, respectively. After oral administration, EP was absorbed 82.0 +/- 6.9 and 98.0 +/- 8.3% under fasted and fed conditions; corresponding mean AUC values were 32.0 +/- 2.0 and 30.8 +/- 3.6 h.microg/mL, respectively. The AUC value for SC-70303 acid (the open lactone form of EP) was lower under fed conditions after cephalic vein infusion, but was greater under fed conditions after portal vein infusion or oral solution administration. The hepatic first-pass effect of EP was 12.6 +/- 6.3% under fasted conditions and 27.1 +/- 6.0% under fed conditions. Pharmacokinetic analysis of EP concentrations after portal vein infusion and oral administration showed that under fed conditions the rate constants for bile excretion and for liver metabolism and urinary excretion were increased while the rate constant for elimination and/or metabolism in the gastrointestinal tract was reduced. In conclusion, the apparent lack of food effect after oral administration was observed because enhanced clearance was compensated by increased absorption.     

Effects of dosing time and schedule on cisplatin-induced nephrotoxicity in rats

Renal dysfunction induced by a single injection of cisplatin depends on the timing of the dose. However, the effects of repeated administration of cisplatin on time-dependent toxicity have not been evaluated despite the fact that in clinical practice high doses are repeatedly injected at intervals or low doses are administered daily. We studied chrononephrotoxicity in rats after weekly or daily cisplatin injections. Weekly high doses (5 mg kg(-1)) or daily low doses (1.2 mg kg(-1)) of cisplatin were injected at four time points (3, 9, 15 and 21 h after the light was turned on (HALO)) for 3 weeks. Changes in body weight after weekly cisplatin administration were independent of the timing of the doses. In the group that received daily cisplatin, the loss in body weight in the 3 HALO group was smaller than in animals receiving injections at 15 and 21 HALO (P < 0.05 and 0.001, respectively). The blood urea nitrogen and serum creatinine levels in the control rats showed a significant circadian change (peak at 15 HALO and trough at 9 HALO), but these parameters were markedly elevated in both trials and their circadian variations were disturbed. In conclusion, cisplatin-induced nephrotoxicity was the lowest at 3HALO compared with other time points of both dose regimens.     

Thienamycin nephrotoxicity. Mitochondrial injury and oxidative effects of imipenem in the rabbit kidney

The nephrotoxic cephalosoprins cephaloridine and cephaloglycin both produce mitochondrial respiratory toxicity in renal cortex. Recent work has provided evidence that this respiratory toxicity is caused by acylation and inactivation of mitochondrial anionic substrate transporters. While cephaloridine also causes significant lipid peroxidative injury in cortical mitochondria and microsomes, cephaloglycin causes little or no oxidative damage under identical conditions. The recently released thienamycin antibiotic, imipenem, like the toxic cephalosporins, produces acute proximal tubular necrosis which can be prevented completely by prior administration of probenecid. The ability of imipenem to block mitochondrial substrate uptake and respiration and produce oxidative changes has not been examined. We therefore evaluated the effects of imipenem in rabbit renal cortex on the following: (1) mitochondrial function [respiration with and uptake of succinate, and uptake of ADP]; and (2) evidence of oxidative change [depletion of reduced glutathione (GSH), production of oxidized glutathione (GSSG), and production of lipid peroxidative injury, as reflected in microsomal conjugated dienes (CDs)]. The mitochondrial effects of 300 mg/kg body wt of imipenem, given i.v. 1 and 2 hr before killing the animals, were comparable to those of the nephrotoxic cephalosporins. There was significant reduction of respiration with, and unidirectional uptake of, succinate at both times, while mitochondrial ADP transport was comparatively unaffected. Imipenem also depleted GSH and increased GSSG and CDs at 1 hr. These effects, however, were considerably smaller than those of a comparably nephrotoxic dose of cephaloridine, and this evidence of oxidative stress had resolved by 2 hr. We conclude that imipenem and the nephrotoxic cephalosporins have similar effects on mitochondrial substrate uptake and respiration, but differ significantly in their production of oxidative injury.     

Complexation hydrogels for oral insulin delivery: effects of polymer dosing on in vivo efficacy

Hydrogels comprised of poly(methacrylic acid) grafted with poly(ethylene glycol) (P(MAA-g-EG)) were characterized and examined for their potential as oral insulin carriers. Insulin loaded polymer (ILP) samples were made using two different polymer formulations. The values for the effective molecular weight between crosslinks, M _e , and the network mesh size, xi, were characterized and increased with increasing pH levels for both formulations. Insulin uptake studies indicated a high insulin loading efficiency for all samples tested, however release was dependent on the amount of insulin loaded. The effect of total polymer dosing was investigated by in situ administration in isolated ileal segments in rats. All ILP samples induced a hypoglycemic effect and an increase in insulin levels, proving that insulin was still biologically active. Insulin dosing amounts were varied by (i) maintaining a constant insulin fraction within an ILP sample while changing the amount of ILP and (ii) by varying the insulin fraction while dosing with the same amount of ILP. The total insulin absorption was dependent on both the amount of the polymer present and the concentration of insulin within an ILP sample, with a maximum relative bioavailability of 8.0%.     

Protective effect of green tea extract on gentamicin-induced nephrotoxicity and oxidative damage in rat kidney

Gentamicin (GM) is an effective aminoglycoside antibiotic against severe infections but nephrotoxicity and oxidative damage limits its long term clinical use. Various strategies were attempted to ameliorate GM nephropathy but were not found suitable for clinical practice. Green tea (GT) polyphenols have shown strong chemopreventive and chemotherapeutic effects against various pathologies. We hypothesized that GT prevents GM nephrotoxicity by virtue of its antioxidative properties. A nephrotoxic dose of GM was co-administered to control and GT-fed male Wistar rats. Serum parameters and enzymes of oxidative stress, brush border membrane (BBM), and carbohydrate metabolism were analyzed. GM increased serum creatinine, cholesterol, blood urea nitrogen (BUN), lipid peroxidation (LPO) and suppressed superoxide dismutase (SOD) and catalase activities in renal tissues. Activity of hexokinase, lactate dehydrogenase increased whereas malate dehydrogenase decreased. Gluconeogenic enzymes and glucose-6-phosphate dehydrogenase were differentially altered in the cortex and medulla. However, GT given to GM rats reduced nephrotoxicity parameters, enhanced antioxidant defense and energy metabolism. The activity of BBM enzymes and transport of Pi declined by GM whereas GT enhanced BBM enzymes and Pi transport. In conclusion, green tea ameliorates GM elicited nephrotoxicity and oxidative damage by improving antioxidant defense, tissue integrity and energy metabolism.     

Protective effect of dietary flaxseed oil on arsenic-induced nephrotoxicity and oxidative damage in rat kidney

Arsenic, a naturally occurring metalloid, is capable of causing acute renal failure as well as chronic renal insufficiency. Arsenic is known to exert its toxicity through oxidative stress by generating reactive oxygen species (ROS). Flaxseed, richest plant based dietary source of ω-3 polyunsaturated fatty acids (PUFAs) and lignans have shown numerous health benefits. Present study investigates the protective effect of flaxseed oil (FXO) on sodium arsenate (NaAs) induced renal damage. Rats prefed with experimental diets (Normal/FXO diet) for 14 days, were administered NaAs (20 mg/kg body weight i.p.) once daily for 4 days while still on the experimental diets. NaAs nephrotoxicity was characterized by increased serum creatinine and blood urea nitrogen. Administration of NaAs led to a significant decline in the specific activities of brush border membrane (BBM) enzymes both in kidney tissue homogenates and in the isolated membrane vesicles. Lipid peroxidation and total sulfhydryl groups were altered upon NaAs treatment, indicating the generation of oxidative stress. NaAs also decreased the activities of metabolic enzymes and antioxidant defence system. Histopathological studies supported the biochemical findings showing extensive damage to the kidney by NaAs. In contrast, dietary supplementation of FXO prior to and alongwith NaAs treatment significantly attenuated the NaAs-induced changes.     

Selenium in the anterior pituitary of rats exposed to sodium selenite: light and electron microscopic localization

Metal selenide accumulations were demonstrated in the anterior pituitary of the rat by a histochemical technique at light and electron microscopic levels. After administration of sodium selenite either by drinking water (2.5 to 15 ppm) or by ip injection (5 to 20 mg/kg body wt), intracellular accumulations were found in secretory granules and lysosomes of the somatotrophs, thyrotrophs, corticotrophs, and the gonadotrophs. The amount of countable deposits increased with increasing doses, whether selenite was given in drinking water or by ip injection. Localization of deposits was independent of route of administration. Following a single ip injection of 5 mg sodium selenite/kg, a steadily increasing amount of visible deposits was seen throughout the first week. After this peak the deposits started to decrease but could still be found after 2 weeks. Selenium may possibly create bonds to endogenous zinc in the anterior pituitary as has been suggested for the brain.     

Protective effects of oral arabic gum administration on gentamicin-induced nephrotoxicity in rats.

Arabic gum (AG) is a complex polysaccharide used as suspending agent. It has been widely used by eastern folk medicine practitioners as a restorative agent and is thought to be an excellent curative for renal failure patients. We therefore tested these folkloric claims using a rat model of gentamicin (GM)-induced nephrotoxicity. AG (7.5g 100ml(-1), in drinking water) was administered orally for 8 days concurrently with GM (80mgkg(-1) per day, i.p.). Estimation of urine volume, serum creatinine and urea concentrations, kidney tissue malondialdehyde (MDA) contents and glutathione (GSH) were carried out after the last dose of GM. Kidneys were also examined for histological changes. GM caused a marked nephrotoxicity as evidenced by significant increases in urine volume (295%), serum creatinine (318%) and urea (258%) and a significant decrease in creatinine clearance (Ccr) (26%). Treatment with AG protected the rats from GM-induced nephrotoxicity as evident by normalisation of these parameters. In addition there was about 187% increase in kidney tissue MDA contents above the control with GM treatment. AG totally prevented the GM-induced rise in kidney tissue contents of MDA. Kidney histology of the tissue from GM-treated rats showed necrosis and desquamation of tubular epithelial cells in renal cortex as well as interstitial nephritis. Whereas it was very much comparable to control when AG was co-administered with GM. In conclusion, AG protected the rats from GM-induced nephrotoxicity, possibly, at least in part through inhibition of the production of oxygen free radicals that cause lipid peroxidation.