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New method for applying zebrafish to evaluate trace sample anti-osteoporosis activity and safety

A technology of osteoporosis and application method, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems associated with toxicity, and achieve the effect of short cycle

Inactive Publication Date: 2018-05-01
JIANGSU PROVINCE INST OF TRADITIONAL CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the anti-osteoporosis screening research of traditional Chinese medicine with zebra larvae, it was found that the drug was metabolized by fish to produce anti-osteoporosis activity, sometimes accompanied by toxic products.

Method used

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  • New method for applying zebrafish to evaluate trace sample anti-osteoporosis activity and safety
  • New method for applying zebrafish to evaluate trace sample anti-osteoporosis activity and safety
  • New method for applying zebrafish to evaluate trace sample anti-osteoporosis activity and safety

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Metabolism, anti-osteoporosis activity and safety of icariin and baohuoside I were evaluated by zebrafish metabolism-effect / toxicity evaluation method.

[0043] 1. Materials and instruments

[0044] Test drug: icariin (National Institute for the Control of Pharmaceutical and Biological Products, batch number 110737-200415).

[0045] Experimental animals: adult zebrafish were provided by the Institute of Model Animals, Nanjing University, from the Tuebingen strain in Germany.

[0046] Reagents: Prednisolone (Suzhou Yake Chemical Reagent Co., Ltd., batch number: YK2012020101); Alizarin Red S (Zhengzhou Siji Chemical Products Co., Ltd., batch number: Sj20110806); Paraformaldehyde (Chengdu Kelong Chemical Reagent Factory, Lot: 20100504); MS-222 (Acros Organics, Lot: A0288328).

[0047] Instruments: ZEISS fluorescence inverted microscope Axioobserver.A1 (Zeiss Optical Instrument International Trade Co., Ltd.); Nikon Aphaphot-2YS2 microscope (Nikon Nikon, Japan); OLYMPUS To...

Embodiment 2

[0071] Zebrafish metabolism-effect / toxicity evaluation method to evaluate the metabolism, anti-osteoporosis activity and safety of total flavonoids of Epimedium.

[0072] 1. Materials and instruments

[0073] Test drug: total flavonoids of Epimedium (including Epimedin A 3.0%, Epimedin B 7.1%, Epimedin C 9.9%, Icariin 29.0%, Baohuoside I 2.4%).

[0074] Experimental animals and instruments are the same as in Example 1.

[0075] 2. Experimental method

[0076] 2.1 Administration

[0077] Embryos were produced by natural mating of adult zebrafish, and the embryos were placed in medium and cultured in a 28°C incubator for 24 hours. Take zebrafish embryos 3pf (3 days after fertilization), put them in a 24-well plate, 8 embryos per well, 2-4 wells in each group, 2mL in each well, and expose to 0.5% DMSO vehicle negative control group and model drug group ( 25 μmol / L prednisolone), 30 μg / mL etidronate disodium positive drug group containing 25 μmol / L prednisolone, 4 doses of epi...

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PUM

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Abstract

The invention relates to a new application method organically combining a model organism zebrafish osteoporosis model, a zebrafish metabolism method and zebrafish toxicity evaluation, and concretely relates to a new method for integrally evaluating zebrafish metabolism-anti-osteoporosis activity / toxicity. The dosage of an experimental sample is less and only needs few mg at most, an experiment iscarried out in a microwell plate by adopting zebrafish embryos fertilized after 1 to 3 days or juvenile fishes, and an experimental period is around one week. The zebrafish osteoporosis model inducedby glucocorticoid is adopted for quickly evaluating trace component anti-osteoporosis activity; in addition, the zebrafish toxicity evaluation is carried out synchronously or combined, a change of a death status or a visceral organ morphology of the zebrafish caused by a test drug is observed; during an anti-osteoporosis activity / toxicity evaluation process, the metabolic transformation of the test drug acted on the zebrafish is further tested, and a metabolite is analyzed by adopting high performance liquid chromatography or a combined mass-spectrometric technique. The invention provides a new technical method for screening an anti-osteoporosis drug, and has a significance on finding safe and effective anti-osteoporosis drugs.

Description

technical field [0001] The present invention relates to a new application method of the organic combination of model organism zebrafish osteoporosis model, zebrafish metabolism method and zebrafish toxicity evaluation, especially the combination of three zebrafish models in terms of anti-osteoporosis activity and safety of microcomponents The application of in vivo evaluation, specifically related to the new method of zebrafish metabolism-anti-osteoporosis activity / toxicity integrated evaluation, that is, using the zebrafish osteoporosis model to evaluate the activity of microcomponents, analyzing metabolites at the same time, and evaluating the effect on zebrafish Fish safety. Background technique [0002] Traditional Chinese medicine is favored in the treatment of osteoporosis, but its liver toxicity has also attracted attention. Some well-known traditional Chinese medicines with definite curative effects, wide clinical use and high attention, such as Zhuanggu Joint Pills ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02
CPCG01N30/02
Inventor 韦英杰贾晓斌刘超陈颖凌洁孙娥宋婕宁青
Owner JIANGSU PROVINCE INST OF TRADITIONAL CHINESE MEDICINE