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Novel method for screening traditional Chinese medicine anti-osteoporosis active component with zebra fish osteoporosis model

A technology for osteoporosis and application methods, which is applied in the analysis of materials, material analysis by optical means, and material inspection products, etc., can solve the problems of large sample consumption, high labor intensity and high cost, and achieves less compound consumption and labor. Strong and efficient separation effect

Inactive Publication Date: 2013-12-04
JIANGSU PROVINCE INST OF TRADITIONAL CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the anti-osteoporosis activity screening of traditional Chinese medicine ingredients, especially micro-components, is still difficult to carry out simply and efficiently. There are two bottlenecks: one is the evaluation model problem: the existing mammalian osteoporosis model takes a long time and requires a large amount of samples , labor-intensive, low efficiency, not suitable for high-efficiency activity screening; the existing osteoblast, osteoclast and tumor cell replacement models have a single action link, which is difficult to reflect the overall view of traditional Chinese medicine; in addition, the cost is high, the experimental conditions are relatively harsh, and primary cells The difficulty of cultivation increases, and it is difficult for ordinary laboratories to carry out; the second is the problem of the number of compounds: due to the complexity of traditional Chinese medicine components, it is difficult to separate enough compounds to meet the requirements of in vivo activity evaluation, and the enrichment of microcomponents and in vivo metabolic components , more difficult to separate
Combining the zebrafish osteoporosis model with modern chromatography can break through the constraints of the evaluation model and the number of compounds in the anti-osteoporosis screening process of traditional Chinese medicine. This aspect of research is still blank in China

Method used

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  • Novel method for screening traditional Chinese medicine anti-osteoporosis active component with zebra fish osteoporosis model
  • Novel method for screening traditional Chinese medicine anti-osteoporosis active component with zebra fish osteoporosis model
  • Novel method for screening traditional Chinese medicine anti-osteoporosis active component with zebra fish osteoporosis model

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Anti-prednisolone-induced zebrafish osteoporosis activity evaluation of component groups captured by high performance liquid chromatography with different retention time periods within 0 to 80 min.

[0055] 1. Materials and instruments

[0056] Drug under test: Dipsacia

[0057] Experimental animals: adult zebrafish (Institute of Model Animals, Nanjing University).

[0058] 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).

[0059] Instruments: ZEISS fluorescence inverted microscope Axioobserver.A1 (Zeiss Optical Instrument International Trade Co., Ltd.); biochemical incubator SPX-80 (Ningbo Haishu Saifu Experimental Instrument Factory); Agilent1100 series high performance liquid chromatography (Agilent Corporation of the Unite...

Embodiment 2

[0087] Anti-prednisolone-induced zebrafish osteoporosis activity evaluation of different components or component groups captured by HPLC within 27-46 min.

[0088] 1. Materials and instruments

[0089] Tested drug: components within 27-46 minutes of dicontinuation

[0090] Experimental animals and instruments are the same as in Example 1.

[0091] 2. Experimental method

[0092] 2.1 Method

[0093] Dipsacus sample preparation: high performance liquid chromatography condition is the same as embodiment one " 2.1 method ", by 27min-46min (such as Figure 5 Shown) the Dipsacus component is further separated and captured by high performance liquid phase. Inject 50 μL of Dipsacus extract (0.1g crude drug / mL) from Example 1 for high-performance liquid chromatography separation, capture the component peaks of Dipsacus at 27-30min, 30-35min, 35-40min, 40-43min and 43-46min respectively , which are denoted as S 1 , S 2 , S 3 , S 4 and S 5 , where S 5 The reference substance w...

Embodiment 3

[0114] Anti-prednisolone-induced zebrafish osteoporosis activity evaluation of different components or component groups captured by HPLC within 24-60 min.

[0115] 1 Materials and Instruments

[0116] Test drug: components within 24-60 minutes of dicontinuation

[0117] Experimental animals and instruments are the same as in Example 1.

[0118] 2 Experimental methods

[0119] 2.1 Method

[0120] Dipsacus sample preparation: high performance liquid chromatography condition is with embodiment one " 2.1 method ", by 24-60min (such as Figure 12 Shown) the Dipsacus component is further separated and captured by high performance liquid phase. Inject 50 μL of the Dipsacaceae extract (0.1g crude drug / mL) in Example 1 for high-performance liquid chromatography separation, and capture the component peaks of Dipsacaceae at 24-40min and 40-60min respectively. According to our previous research (Wei Yingjie, Jia Xiaobin, Fan Hongwei, Peng Yunru, Zhan Wen. China Journal of Traditional...

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Abstract

The invention discloses an improved model organism zebra fish osteoporosis model application method and application of the improved model organism zebra fish osteoporosis model application method in screening the traditional Chinese medicine anti-osteoporosis active component by uniting with the modern chromatography technology. According to the improved model organism zebra fish osteoporosis model application method, the features of high-efficiency separation and analysis of traditional Chinese components of the modern chromatography technology and the united technology of the modern chromatography technology are combined, a target component or component groups are captured or knocked out or authenticated directionally. The component or the component groups which are captured or knocked out are screened in an anti-osteoporosis active mode with the zebra fish osteoporosis model. The traditional Chinese components are separated and analyzed according to the optimized chromatography and mass spectrum conditions. The traditional Chinese components are analyzed according to features of ultraviolet ray absorption, molecular weight and the like. The target component or the component groups are flexibly determined as required, the target component or the component groups of different preserving time periods are collected and thus, the captured or knocked-out traditional Chinese medicine component or the component groups are obtained. After an organic solvent is removed, a proper solvent is used for carrying out dissolution. The zebra fish osteoporosis model which carries out inducing with glucocorticoids rapidly evaluates anti-osteoporosis activity of the traditional Chinese component or the component groups. The modern chromatography and the united technology of the modern chromatography overcome the defects that a traditional chemical component is low in separation speed and a large number of trace components are hard to obtain. The zebra fish osteoporosis model breaks through the bottleneck that the trace components are hard to undergo anti-osteoporosis activity high-efficiency evaluation. The modern chromatography and the united technology are united in an organic mode so as to break through the restraint of an evaluation model and the number of chemical compounds. The anti-osteoporosis pharmacodynamic material basis is provided for screening the traditional Chinese medicine efficiently.

Description

technical field [0001] The invention relates to a new application method for establishing an osteoporosis model with the model organism zebrafish, especially the application of its combined chromatographic technology in the high-efficiency screening of anti-osteoporosis active ingredients of traditional Chinese medicines, in particular to the use of the zebrafish osteoporosis model combined with Modern chromatography such as high performance liquid chromatography or its combined techniques such as mass spectrometry can effectively screen the active ingredients of traditional Chinese medicine for anti-osteoporosis. Background technique [0002] The composition of traditional Chinese medicine is complex, with the characteristics of "multiple components, multiple targets, and overall effect". What method and technology to use to quickly and efficiently clarify the material basis of traditional Chinese medicine is a hot and difficult issue that scholars at home and abroad are co...

Claims

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Application Information

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IPC IPC(8): G01N21/17G01N33/50
Inventor 韦英杰王长梅孙娥贾晓斌谢林宁青景莉君
Owner JIANGSU PROVINCE INST OF TRADITIONAL CHINESE MEDICINE
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