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The Comparisons of the Effects between Enzyme Staining and Biochemical Methods in Drug Safety Evaluation and Ranolazine Genotoxicity Study

Author: ZhangChengMei
Tutor: XieKeQin;LiuZhaoPing
School: Shandong University
Course: Toxicology
Keywords: Hepatotoxicity Low molecular weight fucoidan (LMWF) Beagle dogs Ranolazine (RZ) Mice Genotoxicity
CLC: R927.2
Type: Master's thesis
Year: 2011
Downloads: 12
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Abstract


Drug toxicity is one of the main reasons that lead to drug development failures. In order to avoid toxicity caused by expensive discovered late in the drug development to determine the drug's potential toxicity problems as early as possible is very important. Drug toxicity to the liver and kidney toxicity is the most important. The liver is the main place, is also one of the susceptible target organs of toxicity damage biotransformation and detoxification. According to reports, the drug-induced liver disease or liver damage accounted for 10% -15% of adverse drug reactions. Therefore, early detection of drug toxicity is critical for drug development. This topic is designed on the basis of the experimental study, Beagle dogs as a model, a low component the fucoidan (LowMolecular Weight Fucoidan, LMWF) study, the use of different detection methods to detect changes in the liver sensitive indicators of toxicity early reaction judgment, to analyze the possibility of drug toxicity, to raise vital organs sensitive indicators to predict the toxicity of accuracy. 1 Comparison of enzyme staining and biochemical methods in drug safety evaluation purposes to observe LMWF liver serum biochemical and enzyme histochemistry Beagle dogs. Method (1) Animal grouping and administration: Take 16 healthy Beagle dogs, ♀ ♂ half and half, were randomly divided into vehicle control group (Veh) LMWF low (L, 200 mg / kg / d), medium (M , 800 mg / kg / d), high (H, 3000 mg / kg / d) dose group, n = 4. Gavage, daily morning administration of 1, continuous administration 6d after the withdrawal 1 d. In this way, a total administration of 13 wk. (2) target detection: general symptoms observed daily, body weight was measured every week; detection before the first dose and after the last administration of animal serum biochemical parameters; after the last administration, the liver weighed calculation dirty 1cm3 size organizations coefficient; take the same position in the liver, put frozen for slicing, staining, enzyme histochemical detection; remaining liver with 4% neutral formalin fixed, paraffin-embedded sections, HE staining, optical microscope inspection. Results (1) general symptoms: LMWF each dose group Veh group of animals before the administration and in the pilot phase were not abnormal symptoms, animal activities in good condition, normal diet, drinking water. (2) Weight: The test results showed that the administration period, LMWF different dose groups weight of the animals compared with Veh group did not change significantly (P GT; 0.05). (3) serum biochemical detection: before drug administration and the end of serum biochemical indicators LMWF administration of each group of animals compared with Veh, no significant differences (P gt; 0.05). (4) pathology detection: each dose group animals liver wet weight and organ coefficient Veh group, no significant changes (P gt; 0.05); optical microscopy results showed that, of drug-treated animals liver tissue The morphology found no obvious lesions. (5) liver ATPase (adenosine triphosphatase ATPase) and SDH (Succinate dehydrogenase, SDH) both qualitative observations or quantitative analysis 3000mg/kg/d administration activity decreased significantly compared with Veh group, a statistically significant (P lt; 0.05). Conclusion LMWF administration 13 weeks (1) animals generally good condition, its activities, eating, drinking and weight gain is normal, routine clinical testing no abnormal changes in drug-related. (2) enzyme histochemical detection LMWF 3000 mg / kg / d administered ATPase and SDH activity significantly reduced dose affect liver mitochondrial function, but also that the enzyme were more sensitive than biochemical methods. 2. The Ranolazine genetic toxicity study the genotoxicity of purpose through research Ranolazine (Ranolazine, RZ), to evaluate the safety of medicines. Ames test, mouse bone marrow cell micronucleus test and mouse sperm abnormality test polychromatic erythrocytes, the detection Ranolazine the genetic toxicity. Ames test selection of two ways: the direct method and the metabolic activation method. The test strains of Salmonella typhimurium histidine-deficient strains TA97, TA98, TA100 and TA102 Ranolazine in the trial of six dose groups ((5000,2500,1000,500,250 and 125μg / dish) set the spontaneous revertant control group, and the positive control group; mice bone marrow PCE cell micronucleus test and mouse sperm abnormality test, each test set 3 Ranolazine dose group (222,111 and 55 mg / kg) the negative control group (Veh) and positive control group (CP 40 mg / kg. Results Ranolazine in lt; 5000 μg / dish of the measured dose range, TA97, TA98, TA100 and TA102 four kinds of strains, to join and not to join S9 each dose group of four strains of revertant colony number of the negative control group showed no significant difference (P gt; 0.05), and no dose-response relationship; Ranolazine in the dose range of 55 ~ 222mg/kg, mouse bone marrow cell micronuclei and sperm abnormality rate with the negative control group polychromatic erythrocytes difference between statistical significance (P gt; 0.05). conclusion Ranolazine in the measured dose range, did not show genotoxic effects.

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CLC: > Medicine, health > Pharmacy > Pharmacopoeia Codex ( formulary ),drug identification > Drug identification > Determination of drug content
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