ABSTRACT
D-004, a lipid extract of royal palm (Roystonea regia) fruits that contains a reproducible mixture of fatty acids, has been shown to prevent testosterone and phenylephrine-induced prostate hyperplasia in rodents. This study investigated the long-term oral toxicity of D-004 in rats. Rats from both sexes were randomized into four groups (20 rats sex/group): a control and three treated with D-004 (800, 1500 or 2000 mg/kg/day, respectively). At study completion, rats were sacrificed under anaesthesia. Determinations of blood biochemical and haematological parameters and organ weight were done. Also, necropsy and histopathological studies were performed. Four of 160 rats died before study completion. No clinical signs of toxicity were observed throughout the study. Food and water consumption, bodyweight, blood biochemical and haematological parameters, organ weight ratios and histopathological findings were similar in control and treated groups. The histological lesions found in treated animals are commonly present in this specie and strain according to literature and our historical data. In conclusion, long-term (12 months) oral treatment of rats with D-004 (800-2000 mg/kg/day) did not show evidences of D-004-related toxicity under our conditions. The highest dose tested (2000 mg/kg) was a no-observed adverse effect level in this study.
Subject(s)
Arecaceae/chemistry , Fruit/chemistry , Plant Extracts/toxicity , Administration, Oral , Animals , Drug Administration Schedule , Female , Male , Plant Extracts/administration & dosage , Plant Extracts/chemistry , Rats , Rats, Sprague-DawleyABSTRACT
BACKGROUND AND OBJECTIVE: Benign prostatic hyperplasia (BPH) is a non-malignant enlargement of the prostate that results in obstructive lower urinary tract symptoms. Saw palmetto (Serenoa repens), the dwarf American palm (Arecaceae family), is commonly used to treat BPH. The Cuban royal palm (Roystonea regia) also belongs to the Arecaceae family, and 200-400mg of D-004, a lipid extract from its fruits, administered orally for 14 days has been shown to prevent testosterone- but not dihydrotestosterone-induced prostatic hyperplasia in rats. D-004 (125-250 microg/mL) added to preparations of rat vas deferens caused a marked, dose-dependent and significant inhibition of noradrenaline-induced smooth muscle contraction, a response mediated through alpha(1)-adrenoceptors, and was more effective in these respects than Saw palmetto. However, the in vivo effects of D-004 and Saw palmetto on the hypertensive response induced by noradrenaline were modest (albeit significant), and neither treatment affected resting blood pressure or heart rate in rats. The differential effects of D-004 in in vitro and in vivo models could be related to a differential affinity for adrenoceptor subtypes or to different bioavailabilities in vascular and urogenital targets. Phenylephrine injected into rodents induces prostatic hyperplasia with all the characteristic morphological changes of the condition but does not result in enlargement of the prostate. Therefore, this phenylephrine-induced change in rat prostate tissue is called atypical prostatic hyperplasia. It serves as an in vivo model of prostatic hyperplasia induced by stimulation of alpha(1)-adrenoceptors. The objective of this study was to determine whether D-004 can inhibit induction of atypical prostatic hyperplasia by phenylephrine in rats. METHODS: Rats were randomly distributed into five groups (ten rats/group). One group was a negative control and received oral vehicle only. The other four groups were injected subcutaneously with phenylephrine (2 mg/kg): of these groups, one was a positive control receiving the vehicle, and the other three groups were treated with D-004 or Saw palmetto (both 400 mg/kg) or tamsulosin 0.4 mg/kg. All active treatments were given orally for 28 days. After completion of treatment, rats were placed unrestrained in metabolic cages and micturition studies were performed. The rats were later killed and their prostates removed and weighed. Prostate samples were processed for histological study, with histological changes being assessed according to a scoring system. Bodyweight was measured at baseline and at weekly intervals. RESULTS: Histological examination of positive control rats revealed features of atypical prostatic hyperplasia, with piling-up, papillary and cribiform patterns and budding-out of epithelial cells. Micturition assessment revealed that phenylephrine significantly lowered both the total volume of urine in 1 hour and the volume per micturition; the latter was considered the main efficacy variable. D-004 and Saw palmetto extracts significantly prevented this reduction in volume per micturition by 70.5% and 68.6%, respectively, while tamsulosin totally abolished the reduction in micturition induced by phenylephrine (100% inhibition). Tamsulosin, D-004 and Saw palmetto significantly reduced the histological changes of atypical prostatic hyperplasia induced by phenylephrine by 73.1%, 61.2% and 50.0%, respectively. CONCLUSIONS: Administration of D-004 resulted in marked and significant prevention of phenylephrine-induced impairment of micturition and histological changes in rat prostate. These findings indicate that, in vivo, D-004 effectively opposes these responses to phenylephrine, which are mediated through urogenital alpha(1)-adrenoceptors. In this respect, D-004 was moderately more effective than Saw palmetto, a phytotherapeutic standard used to treat BPH, but less effective than tamsulosin, a selective alpha(1A)-adrenoceptor antagonist.
Subject(s)
Arecaceae/chemistry , Plant Extracts/therapeutic use , Prostatic Hyperplasia/drug therapy , Animals , Disease Models, Animal , Dose-Response Relationship, Drug , Fruit/chemistry , Male , Phenylephrine/chemistry , Plant Extracts/administration & dosage , Plant Extracts/isolation & purification , Prostatic Hyperplasia/chemically induced , Prostatic Hyperplasia/pathology , Rats , Rats, Sprague-DawleyABSTRACT
D-004 is a lipid extract obtained from Cuban royal palm (Rosytonea regia) fruits, consisting of a mixture of fatty acids and esters. D-004 has shown protective effects on prostate hyperplasia induced by testosterone in rodents. We report the results of two studies investigating the acute and subchronic oral toxicity of D004 in rats. Oral acute toxicity of D-004 (2,000 mg/kg) was investigated in Sprague Dawley rats according to the acute toxic class method, and the results showed that D-004 oral acute toxicity was practically absent, being defined as unclassified. In the subchronic study, rats were orally treated with D-004 at 500, 1,000 and 2,000 mg/kg for 90 days. No evidence of treatment-related toxicity was detected. Thus, analysis of body weight gain, clinical observations, blood biochemistry, hematology, organ weight ratios and histopathological data did not show significant differences between control and treated groups. We conclude that D-004 orally administered to rats was safe and that no drug-related toxicity was detected even at the highest dose investigated in both acute and subchronic (2,000 mg/kg) studies. Thus, this dose can be considered as a nonobservable-effect dose in rats.
Subject(s)
Arecaceae/chemistry , Lipids/chemistry , Plant Extracts/toxicity , Toxicity Tests, Acute , Toxicity Tests, Chronic , Administration, Oral , Animals , Female , Fruit/chemistry , Male , No-Observed-Adverse-Effect Level , Prostatic Hyperplasia/prevention & control , Rats , Rats, Sprague-DawleyABSTRACT
Benign prostatic hyperplasia (BPH) is the nonmalignant, uncontrolled growth of prostate gland cells and stroma leading to difficulty in urinating. Lipid extracts from Saw palmetto (Arecaceae) fruits are used to treat BPH. The Cuban royal palm (Roystonea regia) is a member of this family and D-004, a lipid extract from its fruits, prevents prostate hyperplasia (PH) induced with testosterone, as opposed to dihydrotestosterone, in rodents. This study investigated whether D-004 could prevent the histological features of testosterone-induced PH in rats. Rats were distributed into six groups (10 rats per group): A negative control group receiving subcutaneous injections of soy oil and treated with vehicle, and five groups injected subcutaneously with testosterone and treated with the vehicle (positive control), D-004 (100, 200 and 400 mg/kg) or Saw palmetto (400 mg/kg). Treatments were given orally for 14 days. At sacrifice, prostates were removed and processed for light microscopy. The histopathological findings of PH were assessed according to a score-chart protocol. D-004 200 and 400 mg/kg, but not 100 mg/kg, significantly and moderately in a dose-dependent manner prevented prostate enlargement and the testosterone-induced histological changes. Compared with positive controls, D-004 200 and 400 mg/kg inhibited prostate size increases and the histological score up to 56.1% and 60.7%, respectively, while Saw palmetto 400 mg/kg reduced such variables by 45.8% and 49.0%, respectively. The effects of D-004 400 mg/kg on the histological changes, not on prostate size, were greater (p < 0.05) than those of Saw palmetto. D-004 and Saw palmetto did not affect body weight values. In conclusion, D-004 200 and 400 mg/kg administered orally for 14 days prevented the increase of prostate size and the testosterone-induced histological changes in rats, its effects being comparable or mildly better than those of Saw palmetto. These results extend previous data showing preventive effects of D-004 on testosterone-induced prostate enlargement with in rodents, and further studies are required to explore the mechanisms underlying such effects.
Subject(s)
Arecaceae , Phytotherapy , Plant Extracts/administration & dosage , Prostate/drug effects , Prostatic Hyperplasia/pathology , Prostatic Hyperplasia/prevention & control , Administration, Oral , Animals , Dose-Response Relationship, Drug , Intubation, Gastrointestinal , Male , Phytotherapy/methods , Plant Extracts/therapeutic use , Prostate/cytology , Prostate/pathology , Prostatic Hyperplasia/chemically induced , Rats , Rats, Sprague-Dawley , Testosterone/administration & dosageABSTRACT
Osteoporosis is characterized by reduced bone mass, abnormal bone architecture and increased fracture risk. Ovariectomy impairs bone mass and metabolism in rats and ovariectomized rats are considered as a suitable model of postmenopausal osteoporosis. Mevalonate is required for producing lipoids that are important in osteoclast activity and thus drugs affecting mevalonate production can prevent bone loss in rodents. Policosanol is a cholesterol-lowering drug isolated from sugar cane wax that inhibits cholesterol biosynthesis through an indirect regulation of hydroxymethylglutaryl coenzyme A (HMG-CoA) reductase activity. The purpose of this study was to determine whether policosanol could prevent bone loss in the bones of ovariectomized rats by comparing its effects with those induced by estradiol. Sprague Dawley female rats were randomly distributed in four groups: a sham-operated group treated with Tween/H2O vehicle and three groups of ovariectomized rats treated with 17beta-estradiol (30 microg/kg/day) or policosanol (50 and 200 mg/kg/day), respectively, for 3 months. At treatment completion the rats were sacrificed, their bones removed and variables of bone resorption and formation were investigated by histomorphometry. Ovariectomy increased trabecular separation but diminished the number and thickness of trabecules. Estradiol and policosanol prevented these effects compared with ovariectomized controls. Both treatments also prevented an increase in the number of osteoclasts and their surface area induced by ovariectomy. Estradiol, but not policosanol, significantly prevented an increase of osteoblast surface area compared with ovariectomized controls. In conclusion, policosanol prevented bone loss and decreased bone resorption in ovariectomized rats, suggesting that it should be potentially useful in preventing bone loss in postmenopausal women.
Subject(s)
Fatty Alcohols/pharmacology , Osteoporosis/prevention & control , Ovariectomy , Administration, Oral , Animals , Disease Models, Animal , Drug Administration Schedule , Drug Evaluation, Preclinical/methods , Estradiol/administration & dosage , Estradiol/pharmacokinetics , Fatty Alcohols/administration & dosage , Fatty Alcohols/chemistry , Female , Humans , Injections, Subcutaneous , Intubation, Gastrointestinal , Osteoclasts/drug effects , Osteoporosis/drug therapy , Plant Extracts/administration & dosage , Plant Extracts/chemistry , Rats , Rats, Sprague-DawleyABSTRACT
D-003 is a mixture of very high molecular weight aliphatic primary acids purified from sugar cane (Saccharum officinarum, L) wax, in which the most abundant component is octacosanoic acid. Experimental studies have shown that D-003 not only shows cholesterol-lowering and antiplatelet effects, but also offers strong protection against plasma lipoprotein oxidation. Previous studies demonstrated that D-003 protected against the histological changes characteristic of Cl4C-induced hepatic injury in rats. The aim of the present study was to investigate the effects of D-003 in acute hepatotoxicity induced by paracetamol in rats. Male Sprague Dawley rats were randomly distributed in two experimental series of three experimental groups as follows: group 1--positive control rats (paracetamol-treated); groups 2 and 3--rats with liver damage induced by paracetamol and treated with D-003 at 5 and 25 mg/kg, respectively, and which also received paracetamol to induce liver injury. In experimental series 1, animals received paracetamol orally (600 mg/kg). In series 2, paracetamol was administered through the intraperitoneal route (200 mg/kg). Eighteen hours after paracetamol dosing, rats were anesthetized with ether and livers were removed for histopathological studies. In the two experimental series, D-003 at 5 and 25 mg/kg significantly (p < 0.01) decreased the percentage of turgent cells and hepatocytes with necrosis and increased the percentage of normal hepatocytes with respect to positive controls in a dose-dependent manner. Necrotic areas and inflammatory infiltrates were observed in the liver of nine out of ten (90%) positive controls. In turn, D-003 dramatically reduced both necrotic areas and inflammatory infiltrate and was present in only one out of ten (10%) animals treated in the two experimental series. No histological alterations in liver sections of negative controls were found. D-003 protected against the histological changes characteristic of paracetamol-induced hepatic injury in rats, in which the process of lipid peroxidation plays a major role. The relationship between this protective action of D-003 in this experimental model and its antioxidant effects needs to be further investigated before definitive conclusions are drawn.
Subject(s)
Acetaminophen/toxicity , Fatty Acids/therapeutic use , Liver/drug effects , Phytotherapy , Saccharum/chemistry , Waxes/chemistry , Administration, Oral , Animals , Drug Administration Schedule , Drug Evaluation, Preclinical , Fatty Acids/administration & dosage , Fatty Acids/isolation & purification , Liver/pathology , Male , Molecular Weight , Plant Preparations , Rats , Rats, Sprague-DawleyABSTRACT
D-003 is a mixture of very high molecular weight aliphatic primary acids purified from sugar cane (Saccharum officinarum, L.) wax, in which octacosanoic acid is the most abundant component. Previous experimental studies have shown that D-003 not only shows cholesterol-lowering and antiplatelet effects, but also offers strong protection against plasma lipoprotein oxidation. Acute hepatotoxicity induced by CCL4 in rats has been related to an increased rate of lipid peroxidation, and different antioxidant compounds have been revealed to be effective in this model. The aim of this study was to investigate the effects of D-003 in acute hepatotoxicity induced by CCL4 in rats. Male Sprague Dawley rats were randomly distributed in four experimental groups as follows: group 1: negative control rats; group 2: positive control rats (CCL4-treated); groups 3 and 4 rats with liver damage induced by CCL4 and treated with D-003 at 25 and 100 mg/kg, respectively. Acute liver injury was induced by CCL4 suspended in olive oil and intraperitoneally administered at 1 ml/kg. Eighteen hours after CCL4 dosing, the rats were anesthetized with ether and their livers were removed for histopathological studies. D-003 at 25 and 100 mg/kg significantly (p < 0.01) decreased the percentage of ballooned cells and hepatocytes with lipidic inclusions and increased the percentage of normal hepatocytes compared with that in positive controls in a dose-dependent manner. The percent inhibitions of the occurrence of ballooned cells and hepatocytes with lipids were marked (75% and 50%, respectively) with the high dose (100 mg/kg). The percent of turgent hepatocytes was also significantly reduced compared with that in positive controls, but this effect was not dose-dependent. No histological alterations in the liver sections of negative controls were found. Necrotic areas and inflammatory infiltrate were observed in the liver of 7/8 (87.5%) of positive controls. In turn, D-003 dramatically reduced both necrotic areas and inflammatory infiltrate and was present in only 1/8 (12.5%) animals treated with D-003 25 mg/kg and in none (0%) of the animals treated with 100 mg/kg. D-003 protected against the histological changes characteristic of CCL4-induced hepatic injury in rats, in which the process of lipid peroxidation plays the main role. The relationship between this protective action of D-003 on this experimental model and its antioxidant effects needs to be further investigated before definitive conclusions are drawn.
Subject(s)
Fatty Acids/pharmacology , Liver/drug effects , Saccharum , Animals , Antioxidants/pharmacology , Antioxidants/therapeutic use , Carbon Tetrachloride/pharmacology , Chemical and Drug Induced Liver Injury , Fatty Acids/administration & dosage , Hepatocytes/drug effects , Lipid Peroxidation/drug effects , Liver/injuries , Liver Diseases/drug therapy , Male , Phytotherapy , Rats , Rats, Sprague-DawleyABSTRACT
Policosanol is a mixture of higher aliphatic primary alcohols isolated from sugar cane wax, whose main component is octacosanol. An inhibitory effect of policosanol on platelet aggregation and cerebral ischemia in animal models has been reported. Thus, the objective of the present study was to evaluate the effect of policosanol on cerebral ischemia induced by unilateral carotid ligation and bilateral clamping and recirculation in Mongolian gerbils. Policosanol (200 mg/kg) administered immediately after unilateral carotid ligation and at 12- or 24-h intervals for 48 h significantly inhibited mortality and clinical symptoms when compared with controls, whereas lower doses (100 mg/kg) were not effective. Control animals showed swelling (tissue vacuolization) and necrosis of neurons in all areas of the brain studied (frontal cortex, hippocampus, striatum and olfactory tubercle), showing a similar injury profile. In the group treated with 200 mg/kg policosanol swelling and necrosis were significantly reduced when compared with the control group. In another experimental model, comparison between groups showed that the brain water content of control gerbils (N = 15) was significantly higher after 15 min of clamping and 4 h of recirculation than in sham-operated animals (N = 13), whereas policosanol (200 mg/kg) (N = 19) significantly reduced the edema compared with the control group, with a cerebral water content identical to that of the sham-operated animals. cAMP levels in the brain of control-ligated Mongolian gerbils (N = 8) were significantly lower than those of sham-operated animals (N = 10). The policosanol-treated group (N = 10) showed significantly higher cAMP levels (2.68 pmol/g of tissue) than the positive control (1.91 pmol/g of tissue) and similar to those of non-ligated gerbils (2.97 pmol/g of tissue). In conclusion, our results show an anti-ischemic effect of policosanol administered after induction of cerebral ischemia, in two different experimental models in Mongolian gerbils, suggesting a possible therapeutic effect in cerebral vascular disorders.
Subject(s)
Brain Ischemia/drug therapy , Cyclic AMP/analysis , Fatty Alcohols/therapeutic use , Platelet Aggregation Inhibitors/therapeutic use , Animals , Brain Ischemia/pathology , Constriction , Disease Models, Animal , Drug Evaluation, Preclinical , Female , GerbillinaeABSTRACT
Clinical evaluation of 286 asthmatic women showed 15.5% of those who improved clinically had contact with kerosene, while 43.9% of those who failed to improve used kerosene as fuel for cooking. In 16 women the onset of asthma occurred soon after they began to use kerosene. Kerosene can cause and aggravate asthma.
Subject(s)
Asthma/etiology , Kerosene/adverse effects , Petroleum/adverse effects , Asthma/immunology , Asthma/pathology , Environmental Exposure , Female , Humans , Smoke/adverse effectsABSTRACT
Guinea pigs were exposed to kerosene aerosols or to smoke produced from kerosene under conditions approximating those in a kitchen. They were compared to controls exposed to saline aerosols or to atmospheric air. Both types of kerosene exposure engendered aortic plaques, resembling those seen in atherosclerosis, and changes in levels of blood lipids. The results suggest that chronic exposure to kerosene, a domestic fuel widely used in many countries, may have important toxic effects in addition to the pulmonary effects that have been reported by others.
Subject(s)
Aorta/drug effects , Kerosene/toxicity , Petroleum/toxicity , Animals , Aorta/pathology , Cholesterol/blood , Guinea Pigs , Male , Staining and LabelingABSTRACT
Guinea pigs were exposed to kerosene aerosols or to smoke produced from kerosene under conditions approximating those in a kitchen. They were compared to controls exposed to saline aerosols or to atmospheric air. Both types of kerosene exposure engendered aortic plaques with fibrous tissue, collagen, and elastic fibers embedded in abundant glycosaminoglycans-rich ground substance, interspersed in which are smooth muscle cells resembling those seen in atherosclerosis, and changes in levels of blood lipids. The results suggest that chronic exposure to kerosene, a domestic fuel very widely used in many countries, may have important toxic effects in addition to the pulmonary effects that have been reported by others.
Subject(s)
Aerosols/toxicity , Aorta/pathology , Kerosene/toxicity , Petroleum/toxicity , Administration, Inhalation , Animals , Aorta/drug effects , Cholesterol/blood , Cholesterol, HDL/blood , Guinea Pigs , Male , Microscopy, ElectronABSTRACT
Guinea pigs were exposed to kerosene aerosol and compared to controls exposed to atmospheric air. Kerosene exposure induced cytologic and biochemical changes in the pulmonary washings of guinea pigs. Our results suggest that in chronic exposure to kerosene lung washings can be used as a rapid screening tool to detect the inflammatory response in the lungs.
Subject(s)
Kerosene/toxicity , Lung Diseases/chemically induced , Lung/drug effects , Petroleum/toxicity , Aerosols , Alkaline Phosphatase/analysis , Animals , Female , Guinea Pigs , Inflammation , Lung/pathology , Lung Diseases/pathology , Macrophages/enzymology , Macrophages/pathology , Male , Neutrophils/enzymology , Neutrophils/pathology , Therapeutic IrrigationABSTRACT
We undertook a study to determine if pre-exposure to kerosene smoke enhances airway sensitization to egg albumin in the guinea pig. Kerosene vapor inhalation for 15 days, 1 hour daily, in similar conditions to which some housewives who use kerosene as cooking fuel are exposed elicited tracheal damage characterized by signs of dysplasia and inflammatory infiltrate. When these animals were exposed to egg albumin aerosol there was an increase in the antialbumin antibody blood titer and an increased response to egg albumin in the isolated tracheal preparation (Schultz-Dale reaction), We conclude that the airway damage elicited by inhalation of kerosene vapor increase antigen absorption and thereby antibody formation.
Subject(s)
Kerosene/toxicity , Ovalbumin/immunology , Petroleum/toxicity , Smoke/adverse effects , Trachea/drug effects , Tracheal Diseases/chemically induced , Aerosols , Animals , Antibodies/analysis , Antibody Formation/drug effects , Atmosphere Exposure Chambers , Female , Guinea Pigs , Inflammation , Injections, Intraperitoneal , Ovalbumin/administration & dosage , Trachea/immunology , Tracheal Diseases/immunology , Tracheal Diseases/pathologyABSTRACT
It's well known that there exists a high correlation between daily usage of Kerosene and the appearance of dyspnea in healthy humans and in asthmatic patients. Our aim is to study the histological alterations of the respiratory tract of guinea pigs submitted to Kerosene aerosol. It was administered to male guinea pigs fifteen minutes daily for a month. Fragments of trachea and lungs were processed for histological studies. Erosion of tracheal epithelium and inflammatory infiltration were observed. Lungs presented with thickening of the interalveolar septa. The eosinophilic infiltration may represent an immunological response resembling reactions of immediate hypersensitivity. The morphological alterations may be induced by toxic products of Kerosene such as sulphur impurities that act as mucosal irritants which damage defense mechanisms of the organism.