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Method for screening effective sites of flavonoid substances in golden camellia acting on nasopharyngeal carcinoma

A screening method, the technology of Camellia chrysalis, applied in biochemical equipment and methods, microbial determination/testing, biological testing, etc., can solve problems such as difficult to determine anti-tumor active ingredients and complex ingredients

Inactive Publication Date: 2015-04-01
SCHOOL OF MEDICINE JIAYING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Existing literature research is relatively general (alcohol extract or water extract), and the components of the research object are very complex, so it is difficult to determine the real anti-tumor active ingredients and related effective sites

Method used

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  • Method for screening effective sites of flavonoid substances in golden camellia acting on nasopharyngeal carcinoma
  • Method for screening effective sites of flavonoid substances in golden camellia acting on nasopharyngeal carcinoma
  • Method for screening effective sites of flavonoid substances in golden camellia acting on nasopharyngeal carcinoma

Examples

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Effect test

Embodiment 1

[0037] Preparation of golden camellia tea concentrate: pick fresh golden camellia leaves in spring, after screening, washing, crushing and other processes, 20 kg of crushed golden camellia leaves are added; 55L of ethanol (above 95%) is added, and extracted by Soxhlet extractor for 5-6h to obtain extract A; add about 45L of acetone (above 95%) to the residue after extraction, and ultrasonicate for about 1.5h at 40-60°C to obtain extract B; combine extracts A and B, and spin The acetone or ethanol solvent was removed by rotary evaporation with an evaporator to obtain about 2.0 L of Camellia japonica concentrate in the organic phase.

[0038] The preparation process of ferroferromagnetic particles-PAMAM composite material: under nitrogen protection, at pH 1.7, a mixture of ferric chloride solution (0.085 mol / L) and ferrous sulfate (0.05 mol / L) was prepared. Then, 1.5 mol / L ammonia solution was added dropwise to the mixture, and vigorously stirred until pH=9. The ferroferromagne...

Embodiment 2

[0051] Preparation of Camellia camellia flower concentrate: Pick fresh golden camellia flowers in autumn, after screening, washing, crushing and other processes, 10 kg of crushed camellia camellia flowers; add 20L of ethanol (above 95%), and extract with Soxhlet extractor After 5-6 hours, extract A is obtained; add about 15L of acetone (above 95%) to the residue after extraction, and sonicate for about 1.5 hours at 40-60°C to obtain extract B, combine extracts A and B, and use a rotary evaporator The acetone or ethanol solvent was removed by rotary evaporation to obtain about 1.0 L of Camellia japonica flower concentrate in the organic phase.

[0052] The preparation process of ferroferromagnetic particles-PAMAM composite material: under nitrogen protection, at pH 1.7, a mixture of ferric chloride solution (0.085 mol / L) and ferrous sulfate (0.05 mol / L) was prepared. Then, 1.5 mol / L ammonia solution was added dropwise to the mixture, and vigorously stirred until pH=9. The ferr...

Embodiment 3

[0068] Preparation of Camellia Camellia Fruit Concentrate: Picking Camellia Camellia fruit in December, after screening, washing, crushing, vacuum drying and other processes, 20 kg of crushed Camellia Camellia fruit was added; 50L of ethanol (95% or more) was added, and the Extract with the extractor for 5-6 hours to obtain extract A; add about 35L of acetone (above 95%) to the residue after extraction, and sonicate for about 1.5 hours at 40-60°C to obtain extract B, combine extracts A and B, and use The acetone or ethanol solvent was removed by rotary evaporation with a rotary evaporator to obtain about 2.0 L of Camellia camellia fruit concentrate in the organic phase.

[0069] The preparation process of ferroferromagnetic particles-PAMAM composite material: under nitrogen protection, at pH 1.7, a mixture of ferric chloride solution (0.085 mol / L) and ferrous sulfate (0.05 mol / L) was prepared. Then, 1.5 mol / L ammonia solution was added dropwise to the mixture, and vigorously s...

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Abstract

The invention discloses a method for screening effective sites of flavonoid substances in golden camellia acting on nasopharyngeal carcinoma. The method comprises the following steps: extracting flavonoid substances in golden camellia for acting on nasopharyngeal carcinoma cell strains such as CNE-1, CNE-2, C666-1 and the like cultured in vitro and taken as models, discovering that flavonoid-like substances in a golden camellia extract can be used for inhibiting nasopharyngeal carcinoma cell proliferation and inducing apoptosis of nasopharyngeal carcinoma cells by applying a laser confocal inverted microscope, a fluorescence labelling mRNA differential display technology, a proteomics method, matrix-assisted time of flight mass spectroscopy and other technologies, also performing location detection on the expression of genes such as bcl-2, notch1 and the like in nasopharyngeal carcinoma, and revealing the effective sites of the flavonoid substances in golden camellia in inhibition of nasopharyngeal carcinoma activity from the change of protein levels. According to the method disclosed by the invention, deep research can also be performed by virtue of differentially-expressed proteins, action mechanisms of the proteins can be clarified, and meanwhile, the proteins can be possibly taken as action target molecules of medicines for clinically treating nasopharyngeal carcinoma.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for screening the effective sites of flavonoids in camellia japonica on nasopharyngeal carcinoma, which is based on the nasopharyngeal carcinoma cell line cultured in vitro as a model, through differential display reverse transcription polymerization Enzyme chain reaction technology (DDRT-PCR), proteomics methods and other technologies were used to observe the effect of flavonoids in Camellia japonica extract on inhibiting the proliferation, inducing apoptosis and protein changes of nasopharyngeal carcinoma cells. Effective sites of flavonoids in Camellia japonica on nasopharyngeal carcinoma. Background technique [0002] Golden camellia belongs to the family Theaceae and the genus Camellia, and is one of the eight national first-class protected plants in my country. It is recorded in "Compendium of Materia Medica": "Camellia is produced in the south..., and there are also n...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/02C12Q1/68G01N33/68
Inventor 程金生
Owner SCHOOL OF MEDICINE JIAYING UNIV
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