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Method for screening pancreatic lipase restrainer from traditional Chinese medicine

A technology of pancreatic lipase and screening method, which is applied in the directions of biochemical equipment and methods, microbial determination/inspection, material inspection products, etc., and can solve the problems of complex composition and unclear mechanism of action.

Active Publication Date: 2008-02-27
NATURAL MEDICINE INST OF ZHEJIANG YANGSHENGTANG
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  • Application Information

AI Technical Summary

Problems solved by technology

However, traditional Chinese medicine often has complex ingredients and its mechanism of action is not clear.

Method used

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  • Method for screening pancreatic lipase restrainer from traditional Chinese medicine
  • Method for screening pancreatic lipase restrainer from traditional Chinese medicine
  • Method for screening pancreatic lipase restrainer from traditional Chinese medicine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment 1 screens trypsin inhibitor from Chinese medicine

[0022] Experimental Materials:

[0023] Pancrelipase: cloned, expressed and purified in our laboratory. The purity reaches 95%, and the SDS-PAGE electrophoresis pattern is shown in Figure 1.

[0024] Standards were purchased from sigma

[0025] Reaction substrate: resorufin glyceride (1,2-0-dilaudyl-rac-glycero-3-glutaric acid resorufin ester), purchased from sigma

[0026] Positive control: product from Xenical Roche

[0027] Solid phase carrier: Costar 96-well enzyme plate

[0028] Experimental instrument: microplate reader: Perkin-Blmer HTS7000 plus

[0029] Plate washer: Biorad 1575

[0030] Constant temperature incubator: Shanghai Jinghong DNP-9082

[0031] Fully Automatic Accelerated Solvent Extractor: Dionex ASE-300

[0032] Vacuum Freeze Centrifugal Dryer: Thermo Savant SVC 100H

[0033] High-speed traditional Chinese medicine grinder: Zhejiang Dapeng LD-02

[0034] Implementation steps

...

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Abstract

The invention discloses a selective method for selecting the pancrelipase inhibitor from Chinese traditional medicine. The method is to enclose the pancrelipase on the Millipore enzyme standard board; by adding the activator, the pancrelipase can be transferred into the active open structure; after washing, the Chinese traditional medicine extraction is added to culture to remove the uncombined or weakly combined things; after washing, the substrate is added to react and detect the fluorescence intensity of product and compute the inhibiting percent of sample. The method can be used to select the Chinese traditional medicine extraction and decrease the false masculine result made by the surfactant and nonspecific thing in sample to improve the selective hit rate.

Description

technical field [0001] The invention relates to a method for screening pancrelipase inhibitors from traditional Chinese medicines. Background technique [0002] Obesity refers to a disease caused by excessive accumulation of body fat due to energy intake exceeding expenditure. According to the statistics of the World Health Organization: there are 300 million obese people in the world, of which 22 million are children, and 1.1 billion are overweight. Obesity is related to the occurrence of various diseases, and is a risk factor for diabetes, cardiovascular and cerebrovascular diseases, high blood pressure, insulin resistance, gallstones, and certain cancers. In addition, the resulting health problems include reproductive and sexual dysfunction, Diseases of weight-bearing joints, mental disorders, accidental injuries. Therefore, obesity has become a global public health problem. With the increasing incidence of obesity, there is an urgent need for new, effective and safe t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53G01N21/64C12Q1/25
Inventor 薛莲洪孟学于军平汪家权殷秀飞廖玉华胡柳柴婴蕾冯芳余琴芳赵民军周天浩
Owner NATURAL MEDICINE INST OF ZHEJIANG YANGSHENGTANG
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