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Screening method of flavonoid substances in camellia nitidissima to nasopharyngeal darcinoma acted effective bits

A screening method and technology for camellia tea, which are applied in the fields of compound screening, drug screening, biochemical equipment and methods, etc., can solve the problems of difficult to determine tumor-inhibiting active ingredients, complex ingredients, etc.

Inactive Publication Date: 2019-04-02
程金生
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The 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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  • Screening method of flavonoid substances in camellia nitidissima to nasopharyngeal darcinoma acted effective bits
  • Screening method of flavonoid substances in camellia nitidissima to nasopharyngeal darcinoma acted effective bits
  • Screening method of flavonoid substances in camellia nitidissima to nasopharyngeal darcinoma acted effective bits

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Preparation of golden camellia tea concentrate: pick the fresh golden camellia leaves in spring, after screening, washing, crushing and other processes, 1.0 kg of the crushed golden camellia original leaves; add 1.8L of ethanol (more than 95%), and use Extract with an extractor for 5-6 hours to obtain extract A; add about 1.5 L of acetone (above 95%) to the residue after extraction, and sonicate at 40-60°C for about 1.5 hours to obtain extract B, and combine extracts A and B, The acetone and ethanol solvents were removed by rotary evaporation with a rotary evaporator to obtain about 0.26 L of Camellia camellia concentrate in the organic phase.

[0036] Preparation process of ferroferromagnetic particle-PAMAM composite material: under nitrogen protection, at pH 1.7, a mixture of ferric chloride solution (0.085mol / L) and ferrous sulfate (0.05mol / L) was prepared. Then, a 1.5 mol / L ammonia solution was added dropwise to the mixture, and vigorously stirred until pH=9. The f...

Embodiment 2

[0050] Preparation of camellia camellia flower concentrate: picking fresh golden camellia flowers in autumn, screening, washing, crushing and other processes, 0.9 kg of crushed camellia camellia flowers; adding 1.9L of ethanol (more than 95%), using a Soxhlet extractor Extract for 5-6 hours to obtain extract A; add about 1.3L 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 rotating The acetone or ethanol solvent was removed by rotary evaporation with an evaporator to obtain about 0.25 L of Camellia camellia flower concentrate in the organic phase.

[0051]Preparation process of ferroferromagnetic particle-PAMAM composite material: under nitrogen protection, at pH 1.7, a mixture of ferric chloride solution (0.085mol / L) and ferrous sulfate (0.05mol / L) was prepared. Then, a 1.5 mol / L ammonia solution was added dropwise to the mixture, and vigorously stirred until pH=9. Th...

Embodiment 3

[0068] Preparation of camellia japonica fruit concentrate: picking chrysanthemum camellia fruit in December, screening, washing, crushing, vacuum drying and other processes, 1.1 kg of camellia japonica fruit after crushing; adding 1.9L of ethanol (more than 95%), using the Extracted with the extractor for 5-6 hours to obtain extract A; add 1.7L acetone (above 95%) to the residue after extraction, and ultrasonicate for about 1.5h at 40-60°C to obtain extract B, and combine extracts A and B, The acetone and ethanol solvents were removed by rotary evaporation with a rotary evaporator to obtain about 0.3 L of Camellia camellia fruit concentrate in the organic phase.

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

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Abstract

The invention discloses a screening method of flavonoid substances in camellia nitidissima to nasopharyngeal darcinoma acted effective bits. The flavonoid substances in the camellia nitidissima are extracted, nasopharyngeal darcinoma cell strains of CNE-1, CNE-2, C666-1 and the like acting on culture in vitro are used as a model, and techniques of a laser confocal inverted microscope, a fluorescence labelling mRNA differential display technique, a proteomics method, a substrate auxiliary flight time mass spectrum and the like are applied to find that the flavonoid substances in a camellia nitidissima extract can restrain cancer cell growth of nasopharyngeal darcinoma, can induce apoptosis of cancer cells, and can position expression of genes of bcl-2, notch1 and the like to the nasopharyngeal darcinoma. Changes of protein level reveal that the flavonoid substances in the camellia nitidissima can restrain active effective bits of the nasopharyngeal darcinoma. The method can also deeplyresearch the protein of differential expression so as to clarify the action mechanism, and besides, the protein can be used as a target molecule of drugs for clinically treating the nasopharyngeal darcinoma.

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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IPC IPC(8): C12Q1/02C12Q1/6851G01N33/68
CPCC12N2503/02C12Q1/6851G01N33/5011G01N33/68G01N2500/10C12Q2531/113
Inventor 程金生
Owner 程金生
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