Modified zebra fish drug metabolism modeling method

A technique of metabolic model and establishment method, which is applied in the field of establishment of improved zebrafish drug metabolism model, to achieve the integrated effects of high-efficiency enrichment and in vivo anti-osteoporosis activity evaluation

Inactive Publication Date: 2014-03-12
JIANGSU PROVINCE INST OF TRADITIONAL CHINESE MEDICINE
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  • Modified zebra fish drug metabolism modeling method
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  • Modified zebra fish drug metabolism modeling method

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Embodiment 1

[0063] Evaluation of the anti-osteoporotic activity of icariin and its metabolite baohuoside I in a zebrafish metabolism and osteoporosis model combined with chromatography

[0064] 1. Materials and instruments

[0065] Test drug: icariin (National Institute for the Control of Pharmaceutical and Biological Products, batch number 110737-200415);

[0066] Experimental animals: adult zebrafish (Institute of Model Animals, Nanjing University).

[0067] Reagents: prednisolone (Suzhou Yake Chemical Reagent Co., Ltd., batch number: YK2012020101); etidronate disodium (National Institute for the Control of Pharmaceutical and Biological Products, batch number 101174-201001); Alizarin Red S (produced by Zhengzhou Sisi Chemical Ltd., batch number: Sj20110806); paraformaldehyde (Chengdu Kelong Chemical Reagent Factory, batch number: 20100504); MS-222 (Acros Organics, batch number: A0288328). Acetonitrile and formic acid were chromatographically pure (US TEDIA Company), normal saline (Chi...

Embodiment 2

[0103] Zebrafish metabolism and osteoporosis model combined with chromatography to evaluate the anti-osteoporosis activity of epimedin A and its metabolite baohuoside I.

[0104] 1. Materials and instruments

[0105] Test drug: Epimedin A (made in the laboratory, purity ≥ 98%)

[0106] Experimental animals and instruments are the same as in Example 1.

[0107] 2. Experimental method

[0108] 2.1 Enrichment of Metabolites of Epimedin A with Zebrafish

[0109] Zebrafish dosing and sampling methods: Take 10 adult zebrafish, place them in brown bottles containing 30 mL of solution, and divide them into 2 groups on average, with 5 fish in each group. One group was 0.5% DMSO ultrapure water fish group (blank fish group), and the other group was 0.5% DMSO pure aqueous solution fish group of Epimedin A (drug fish group: Epimedin A: 7.0 μg·mL- 1); the remaining drug solution without adding fish was the blank drug group (i.e. 0.5% DMSO ultrapure aqueous solution of Epimedin A 7.0 μg...

Embodiment 3

[0138] Zebrafish metabolism and osteoporosis model combined with chromatography to evaluate the anti-osteoporosis activity of Epimedin C and its metabolite 2''-O-Rhamnosylicariside II.

[0139] 1 Materials and Instruments

[0140] Test drug: Epimedin C (made in the laboratory, purity ≥ 98%)

[0141] Experimental animals and instruments are the same as in Example 1.

[0142] 2 Experimental methods

[0143] 2.1 Enrichment of Metabolites of Epimedin C Using Zebrafish

[0144] Zebrafish dosing and sampling methods: Take 10 adult zebrafish, place them in brown bottles containing 30 mL of solution, and divide them into 2 groups on average, with 5 fish in each group. One group was 0.5% DMSO ultrapure water fish group (blank fish group), and the other group was 0.5% DMSO pure aqueous solution fish group of Epimedin C (drug fish group: Epimedin C: 7.85 μg·mL -1 ); the remaining drug solution without adding fish was the blank drug group (i.e. Epimedin C 7.85 μg·mL -1 0.5% DMSO ultrap...

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Abstract

The invention relates to a modified zebra fish drug metabolism modeling method, which specifically comprises the steps of (1) enriching metabolites by virtue of a zebra fish metabolism model; (2) separating and analyzing the enriched metabolic components of the zebra fish in the last step by using modern chromatography or the hyphenated technique thereof, capturing or knocking out target components or metabolites; (3) collecting zebra fish germ cells; (4) putting zebra fish embryos in three days after fertilization in a culture plate which holds an embryo culture medium and administrating a drug; (5) dyeing the bones of the zebra fish; and (6) detecting and analyzing the dyed bones of the zebra fish. According to the invention, an adolescent zebra fish osteoporosis model is applicable to in-vivo effective evaluation of anti-osteoporosis activity of Chinese herbal medicinal ingredients such as trace ingredients; an adult zebra fish metabolism model is simple in metabolites and efficient in enrichment; the modern chromatography or the hyphenated technique thereof provide powerful guarantee for the separation and analysis of the complex Chinese herbal medicinal system and the metabolic components. The model obtained by the method has the outstanding advantage that the separation and analysis of Chinese herbal medicinal original-form ingredients and the metabolic components are integrated with the in-vivo anti-osteoporosis activity evaluation, and simple and efficient.

Description

technical field [0001] The invention relates to an improved ready-to-use two zebrafish models, combined with the application of chromatographic technology in the efficient evaluation of the anti-osteoporosis activity of microcomponent metabolites, in particular to the use of model organisms zebrafish metabolic model and zebrafish osteoporosis The model combined with modern chromatography such as high performance liquid chromatography or its combined technology such as mass spectrometry can efficiently evaluate the anti-osteoporosis activity of traditional Chinese medicine metabolites. Background technique [0002] The active substances of traditional Chinese medicine include two aspects: the original compound and its biotransformation products and / or metabolites. The metabolic transformation of the drug in the body produces efficacy or toxicity, which is very important for determining the active ingredients and effective effect substances of traditional Chinese medicine. [...

Claims

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Application Information

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IPC IPC(8): A61K49/00G01N21/00G01N1/30G01N30/02
Inventor 韦英杰詹扬孙娥贾晓斌姚楠舒娈
Owner JIANGSU PROVINCE INST OF TRADITIONAL CHINESE MEDICINE
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