Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Ganoderma quality evaluation method based on in-vitro tumor cell culture model

A technology of tumor cells and Ganoderma lucidum, applied in biochemical equipment and methods, measuring devices, and microbial determination/inspection, etc., can solve the problem of lack of microenvironment for cell growth and differentiation, and can not simulate tumor cells and tumor microenvironment well Interaction, the correlation between the clinical application of Ganoderma lucidum and its curative effect is not strong, etc.

Pending Publication Date: 2020-10-27
ZHEJIANG SHOUXIANGU BOTANICAL DRUG INST CO LTD
View PDF5 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The shortcoming of this method is: although utilize UPLC-MS / MS technology to be able to measure rapidly the content of ganoderma acid C2, ganoderma acid A, ganoderma acid G, ganoderma acid B, ganoderma acid A and ganoderma enoic acid D in the ganoderma sample, but this does not It cannot directly reflect the biological activity of each component, nor can it fully reflect the quality of Ganoderma lucidum
The shortcoming of this method is: although the content of ganoderma acid A, ganoderma acid B, ganoderma acid C2 and ganoderma acid G in the Ganoderma lucidum medicinal material can be quickly determined by high performance liquid chromatography, and the content of the total triterpenoids of ganoderma lucidum can be calculated, the content cannot be determined. It directly reflects the biological activity of the total triterpenes of Ganoderma lucidum, and cannot fully reflect the quality of Ganoderma lucidum medicinal materials
The disadvantage of this method is: the detection of the antioxidant activity of Ganoderma lucidum extract can reflect the quality of Ganoderma lucidum medicinal materials to a certain extent, but the correlation between antioxidant activity and the clinical application and curative effect of Ganoderma lucidum is not strong
The disadvantage of this method is that although HPLC can accurately detect the content of adenosine in Ganoderma lucidum capsules, the content of adenosine cannot fully reflect the activity and quality of Ganoderma lucidum capsules.
The two-dimensional (2-Dimensional, 2D) culture model of tumor cell monolayer adherent growth is currently the main method for evaluating the activity of antitumor drugs. This culture model is convenient, economical, and easy to operate, but it lacks the microenvironment for cell growth and differentiation in vivo , cannot well simulate the interaction between tumor cells and the tumor microenvironment in vivo, and there may be some differences with the results of animal experiments and clinical experiments

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Ganoderma quality evaluation method based on in-vitro tumor cell culture model
  • Ganoderma quality evaluation method based on in-vitro tumor cell culture model
  • Ganoderma quality evaluation method based on in-vitro tumor cell culture model

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Example 1: Evaluation of the quality of Ganoderma lucidum based on chemical detection

[0060] Five batches of Ganoderma lucidum and five batches of Ganoderma sinense were provided by Wuyi Shouxiangu Traditional Chinese Medicine Pieces Co., Ltd. According to "Chinese Pharmacopoeia" 2015 edition, the method stipulated under [Content Determination] under Ganoderma lucidum item, the content of polysaccharides, triterpenes and sterols of these 10 batches of Ganoderma lucidum samples were determined: the determination of polysaccharides was carried out by anthrone-sulfuric acid method, Anhydrous glucose was used as the reference substance; triterpenes and sterols were determined by the vanillin-glacial acetic acid method, and oleanolic acid was used as the reference substance. The test results are shown in Table 1.

[0061] It can be seen from Table 1 that the polysaccharide content of 5 batches of Chizhi is 1.19%-1.91%, the content of triterpenes and sterols is 0.85%-1.58%...

Embodiment 2

[0064] Example 2: Quality evaluation of Ganoderma lucidum based on 2D tumor cell culture model

[0065] Get the same red sesame and purple sesame medicinal materials as in Example 1, and evaluate its quality according to the following steps:

[0066] (1) Take 2g each of different batches of ganoderma lucidum medicinal powder (passed through a 40-mesh sieve), add 50mL of water, heat and reflux in a water bath for 1 hour, filter, put the filtrate on a water bath and evaporate to dryness, and obtain a ganoderma lucidum sample to be tested;

[0067] (2) Get each 40 mg of Ganoderma lucidum sample to be tested, and use DMEM medium to prepare a sample solution (concentration 40 mg / mL);

[0068] (3) Take 10 mg of fluorouracil and prepare a positive control solution (concentration 10 mg / mL) with purified water;

[0069] (4) After the colon cancer cell HCT116 was digested with 0.25% trypsin, the tumor cells were diluted and resuspended with DMEM medium (the cell concentration was about...

Embodiment 3

[0073] Example 3: Evaluation of Ganoderma lucidum sample quality based on 3D tumor cell culture model

[0074] Get the same batch of red sesame and purple sesame medicinal materials as in Example 1, and evaluate its quality according to the following steps:

[0075] Steps (1) to (3) are the same as in Example 2.

[0076] (4) Get 2D cultured colon cancer cell HCT116 (2D culture model construction method is the same as in Example 1) and use 1:2 diluted Matrigel hydrogel as matrix material to build a 3D culture model, and use DMEM medium to combine steps (2) and The Ganoderma lucidum sample solution and fluorouracil solution obtained in (3) were diluted to different concentrations (the final concentration was the same as that in step (4) of Example 2), and incubated with HCT116 cells for 48 hours respectively, with 3 replicate wells for each concentration.

[0077] (5) Aspirate the Ganoderma lucidum sample solution and fluorouracil solution, add 90 μL of fresh DMEM medium, then ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to a ganoderma quality evaluation method based on an in-vitro tumor cell culture model. The method comprises the following steps: taking to-be-detected ganoderma samples and positive control products to produce sample solutions and positive control product solutions, and mixing the sample solutions and the positive control product solutions for co-incubation with tumor cellssubjected to 2D culture or 3D culture after the sample solutions and the positive control product solutions are diluted or conducting co-incubation on the sample solutions and tumor cells subjected to2D culture or 3D culture and co-incubation on the positive control product solutions and tumor cells subjected to 2D culture or 3D culture; and adding a cell activity indicator into incubated solutions separately for reactions, detecting the absorbance or the fluorescence intensity of reaction solutions, calculating tumor cell proliferation inhibition rates of the sample solutions, the positive control product solutions and mixed solutions of the sample solutions and the positive control product solutions, calculating corresponding IC50 or growth inhibition rates, and evaluating the ganodermaquality by the inhibition rates or the growth inhibition rates. According to the ganoderma quality evaluation method based on the in-vitro tumor cell culture model, on the basis of the tumor cells cultured in vitro, the quality of ganoderma is evaluated by detecting tumor cell inhibition effects of the ganoderma, detection indexes are directly related to the anti-tumor activity of the ganoderma,and clinical efficacy of the ganoderma can be reflected so that the quality of the ganoderma can be evaluated more comprehensively; and the tumor cells subjected to 3D culture is more similar to in-vivo tumors in situ, so that evaluation results are more accurate, animal ethics is not involved, an analysis process is short, and the analysis cost is low.

Description

[0001] (1) Technical field [0002] The invention relates to a ganoderma quality evaluation method based on an in vitro tumor cell culture model. [0003] (2) Background technology [0004] Quality control of traditional Chinese medicine is of great significance to ensure the safety, effectiveness and consistency of traditional Chinese medicine products, and is the key to the modernization and internationalization of traditional Chinese medicine. With the continuous development of analytical chemistry, pharmaceutical analysis and other related disciplines, the quality evaluation technology of traditional Chinese medicine has made great progress, and gradually formed a quality inspection technology system with chemical detection as the core. etc. to evaluate the quality of traditional Chinese medicine products. However, the chemical composition of traditional Chinese medicine is complex and the effective components are often unknown. It is difficult to measure one or a few inde...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12Q1/18C12Q1/02
CPCG01N33/5011G01N33/5044
Inventor 李振皓潘海涛张国亮李振宇赵建霞周莎莎
Owner ZHEJIANG SHOUXIANGU BOTANICAL DRUG INST CO LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com