| All trans retinoic acid (acid All-trans-retinoic, ATRA) is a derivative of vitamin A, which can be used in the treatment of acute leukemia in clinic, which can obviously promote the differentiation of leukemia cells and inhibit their proliferation. There are studies to confirm that ATRA has played a certain role in the treatment of lung cancer, prostate cancer, breast cancer, glioma, neuroblastoma, and skin cancer. Recent studies have confirmed that ATRA has a good effect on the use of some drugs. However, in the clinical application of ATRA has great side effects and clinical response, including the lips, dry skin with desquamation, gastrointestinal reactions, headache, dizziness, joint pain and other. These side effects and clinical responses greatly limit the clinical application of ATRA. The first part of this study is to observe the effect of ATRA intragastric administration on the nervous system of mice. Determination of all trans retinoic acid in a non desired drug effect, to evaluate the adverse drug reactions or pathophysiological effects observed in ATRA in toxicology or clinical studies, and to study the mechanism of drug adverse reactions. The second and the third part aims to study the intragastric administration of all trans retinoic acid on rat acute and long-term toxicity and safety evaluation of oral ATRA, provide a reference for the safety evaluation of clinical use of drugs. Therefore, to expand the idea for the clinical application of ATRA, and also hope to provide valuable information for future research.Part 1 A general study on the effect of all-trans-retinoic acid on the nervous system of miceOBJECTIVE To investigate the effect of the all-trans-retinoic acid (ATRA) on the nervous system of mice by employment mice spontaneous activity test, coordination test, and the synergistic effect of the threshold dose of sodium pentobarbital test.METHODS ICR mice, half male and half female, were randomly assigned as vehicle, 250.0 mg/kg,50.0mg/kg and 10.0mg/kg ATRA groups according to body weight. After 12 h of fasting (with water), the mice in each group were given the corresponding dose of ATRA respectively by intragastrical administration, dosing volume was 0.02 ml/g. As to mice spontaneous activity test, the mice spontaneous activity was determined before and after administration of ATRA for 60 min,120 min, and 180 min. Mice coordinated test was carried out by determining the residence time of mice on the rotary rod 120 min after administration. The synergistic effect of the threshold dose of sodium pentobarbital and ATRA was analyzed 120 min after administration of sodium pentobarbital. The numbers of sleeping mice were calculated.RESULTS 250.0mg/kg ATRA can significantly reduce the number of spontaneous activity of mice,50.0mg/kg or 10.0mg/kg ATRA was found to reduce the trend of spontaneous activities in 120 min,180 min; 250.0 mg/kg,50.0 mg/kg, or 10.0 mg/kg ATRA had no significant effect on the coordination of mice. ATRA had no synergistic effect with the threshold dose of sodium pentobarbital.CONCLUSION ATRA could reduce the number of spontaneous activity of mice, and had no significant effect on the coordination of mice, and had no synergistic effect with the threshold dose of sodium pentobarbital.Part 2 Experimental study on the maximum dosage of all trans retinoicacid in rats OBJECTIVE Toxic effects of all trans retinoic acid on SD rats were investigated. Preliminary evaluation of the safety of all trans retinoic acid in order to provide a reference for the follow-up study.METHODS The rats were fasted for 8 hours, intragastric administration of 0.02ml/g in the next day, the toxic reaction was observed immediately after administration, giving food after lh, intragastric administration of 0.02ml/g again after 3h, the toxic reaction was observed.14 days later, the rats were dissected and examined by the naked eye. Statistical analysis of weight using T-test method.RESULTS After administration, rats appearance, behavior, reaction, secretion and excretion index showed no abnormality, and after administration for 4 days, in addition to part of the rat eye view visible eyelid bleeding, other indexes showed no abnormality, nor the death of rat. The ATRA group was administered after 3 days, the male and female rats compared with the solvent control group body weight was significantly reduced, after 3 days, the female and male rats have rebounded, the growth rate of male is higher than female rats, the end of the experiment ATRA group of female and male rats compared with solvent control group still lighter. Test the final dissection of rat, the main organs showed no obvious abnormalities.CONCLUSION 4.0g/kg ATRA did not cause the death of rats, there was no significant abnormal effect on rats, but the weight of rats had obvious effects.Part 3 Long term toxicity test of all trans retinoic acid in ratsOBJECTIVE To observe the effect of toxicity, dose and toxic effect, the main target organ, the nature and degree of toxicity, the reversibility of toxicity, and the reversibility of the rats after 1 months of oral administration of all trans retinoic acid; Clear the toxic reaction of the animal dosage, the toxic reaction dose and the safe range; Determination of non-toxic reaction dose and clinical main detection index, to provide reference for clinical drug safety evaluation.METHODS Select the quarantine of 100 rats, weighing 160-180 g, male and female, were randomly divided into 5 groups, respectively, solvent control group, ATRA 750.0mg/kg group, ATRA 250.0mg/kg, ATRA 50.0mg/kg dose group, ATRA 10mg/kg dose group,10mg/kg dose group,20 rats in each group. The rats in each dose group were given the corresponding dosage of the medicine solution, and the volume of the drug was 0.01ml/g, the solvent control group was administered with equal volume of 0.5%CMC-Na solution, continuous administration for 1 months, recovery period was 2 weeks. During this period, the observation index were:symptoms, body weight, food intake, hematology and blood coagulation function tests, blood biochemistry examination, system anatomy, organ coefficient and histopathological examination.RESULTS Compared with the control group, the body weight and daily food intake in administration group were significantly reduced, and the growth rate of body weight during the administration period was significantly slower than that of the solvent control group. The body weight and daily food intake of the administration group in the recovery period were significantly increased. Compared with the solvent control group, the red blood cell count, hemoglobin concentration, red blood cell ratio and red blood cell content of female rats were significantly decreased, in a dosedependent manner. Male rats compared with solvent control group,250.0mg/kg and 50.0mg/kg group of hemoglobin, mean hemoglobin concentration, mean corpuscular hemoglobin concentration was significantly lower, in 50.Omg/kg group the number of reticulocyte, reticulocyte percentage increased significantly. The number of white blood cells and lymphocytes in the 10.0mg/kg group were significantly increased. Part of the activated prothrombin time, rats in the administration group were longer than those in the control group, but only 10.0mg/kg male rats were significantly different. Compared with the control group, the prothrombin ratio of rats in administration group was lower than those in control group. The prothrombin time and prothrombin international normalized ratio of the administration group were higher than those in the normal group, but only 10.0mg/kg male rats were significantly different. The thrombin time of male rats had a tendency to decrease and the dose was consistent with the dose. The 250.0mg/kg group and 50.0mg/kg group of female rats were significantly lower than the control group. There was a significant decrease in total protein and albumin in the female rats during administration in a dosedependent manner. Compared with the control group, the alkaline phosphatase, blood glucose and triglyceride were significantly increased, and the changes of alkaline phosphatase and triglyceride were dose related. There was a significant decrease in albumin content of male rats during administration period, and there was a significant increase in alanine aminotransferase and triglyceride. After two weeks of recovery period, the indexes of male and female rats in each group were better than those in the drug administration period. The organ coefficient, of female rats in 250.0mg/kg group of spleen, liver and kidney, adrenal gland, heart and brain were significantly increased, thymus coefficient decreased significantly; the organ coefficient of female rats in 50.0mg/kg group of liver and adrenal gland increased significantly; the kidney organ coefficient increased significantly, thymus coefficient significantly reduce. The organ coefficients of spleen, liver, kidney, adrenal gland, lung, heart and brain of male rats in the 250.0mg/kg group were significantly increased, and the liver of 50.0mg/kg and 10.0mg/kg group were significantly increased.5 days after administration of 750.0mg/kg rats that have mandibular hair removal, mouth with blood secretions, secretion and locomotor activity decreased, instability of gait, shortness of breath and other symptoms. After 14 days of treatment, the administration group become dying.18 days after administration,250.0mg/kg group rats were given to reduce spontaneous activity, eyelid bleeding, weakness and other symptoms, the female rats did not die, but 4 male rats were dead.50.0mg/kg group and 10.0mg/kg group behavior appearance is more normal. In 250.0mg/kg,50.0mg/kg and 10.0mg/kg three groups, there were hepatic cell edema^ degeneration and necrosis of the duodenal mucosa, and the increase of the spleen megalokaryocyte; 250;0mg/kg dose group of animal thymus appeared cortical cells decreased; the necrosis of the liver and thymus cortex cells appeared in the 250.0mg/kg and 50mg/kg dose group; 250.0mg/kg group also appeared testicular fine tubules atrophy, reduced germ cells, some animals appear hair follicles decreased. In the recovery period, the spleen megakaryocyte of each group were increased.250.0mg/kg group was higher than the solvent control group, 50.0mg/kg group and 10.0mg/kg group were lower,2/3 animal testis of 250.0mg/kg group showed moderate atrophy of seminiferous tubules, spermatogenic cells decreased, 2/8 animal hair follicles decreased. Overall pathological changes compared to the drug administration period has been restored.CONCLUSION All trans retinoic acid high dose has inhibitory effect on bone marrow hematopoietic system in rats; injury of rat liver; cause thrombocytopenia, anemia and hypertriglyceridemia. |
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