Method for analyzing impact on RAW264.7 and K562 cells from IC50 dose delta-tocopherol

A technology of RAW264.7 and analysis method, which is applied in the field of analyzing the influence of IC50 dose δ-tocopherol on RAW264.7 and K562 cells, and can solve problems such as the influence of metabolomics

Active Publication Date: 2017-09-26
SHANDONG NORMAL UNIV
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Problems solved by technology

There are few reports on the metabol...

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  • Method for analyzing impact on RAW264.7 and K562 cells from IC50 dose delta-tocopherol
  • Method for analyzing impact on RAW264.7 and K562 cells from IC50 dose delta-tocopherol
  • Method for analyzing impact on RAW264.7 and K562 cells from IC50 dose delta-tocopherol

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preparation example Construction

[0077] (2) Preparation of cell extract:

[0078] Preparation of cell extracts in the δ-TOH group: RAW264.7 or K562 cells in the corresponding δ-tocopherol IC 50 Cultivate at a certain dose, add cold methanol after culturing to quench cell metabolism, then use methanol chloroform water extraction method for ultrasonic crushing and extraction, and collect the water phase;

[0079] Preparation of cell extract in the blank group: culture RAW264.7 or K562 cells, add cold methanol after culture to quench cell metabolism, then use methanol chloroform water extraction method for ultrasonic crushing and extraction, and centrifuge to collect the water phase;

[0080] In the process of preparing cell extracts, for the tissue structure and components of RAW264.7 or K562 cells, in order to ensure as much information as possible on the metabolites of cells in the δ-TOH group and cells in the blank group, the extraction reagent is preferred What is used is a mixed solvent of methanol, chlor...

Embodiment 1

[0123] 1. Basic method principle

[0124] Determination of the IC of delta-tocopherol on RAW264.7 and K562 cells 50 Dose, the sample was detected by nuclear magnetic resonance spectrometer (specifically, AVANCE III 600MHz full digital superconductor), and the spectrum was processed by Topspin3.2 (Bruker Biospin, Germany). NMR spectra were processed using MestReNova 6.11 (Mestrelab Research, Espain). Afterwards, the data were imported into SIMCA-P+12.0 software (Umetrics Inc., Umea, Sweden). Data were analyzed using Principal Component Analysis (PLS-DA) and Orthogonal Partial Least Squares Discriminant Analysis (OPLS-DA) to look for differential metabolites. Qualitative analysis was performed through the Human Metabolome Database (HMDB) and the Biological Nuclear Magnetic Resonance Database (BMRB) to determine the species names of the differential metabolites. The identified differential metabolites were imported into the Kyoto Encyclopedia of Genes and Genomes (KEGG) databa...

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Abstract

The invention discloses a method for analyzing the impact on RAW264.7 and K562 cells from IC50 dose delta-tocopherol on the basis of <1>H NMR metabonomics. The method comprises the following steps: (1) determining IC50 dose of delta-tocopherol to the RAW264.7 or K562 cells; (2) preparing a cell extract; (3) performing magnetic resonance imaging pretreatment; (4) performing <1>H magnetic resonance imaging detection; (5)screening differential metabolites; and (6)performing metabolism pathway analysis. According to the method, the H-nuclear magnetic resonance technology is combined with the metabonomics to study the impacts on differential metabolites and metabolic pathways of different cells, and the analytic method for studying the sub-toxicity of delta-tocopherol to the RAW264.7 and K562 cells is provided.

Description

technical field [0001] The present invention involves 1 Analysis of IC by H NMR metabolomics method 50 The method of the effect of dose delta-tocopherol on RAW264.7 and K562 cells belongs to the metabolomics analysis at the cellular level. Background technique [0002] Biological Activity of Vitamin E: [0003] Vitamin E is an essential molecule to maintain normal metabolism and various physiological functions of the human body. Moreover, vitamin E cannot be produced by the human body itself, and needs to be ingested from the outside world through food. Foods rich in vitamin E include almonds, hazelnuts, soybeans, and avocados. Vitamin E, also known as tocopherol, can be divided into the following eight types of α-TOH, β-TOH, γ-TOH, and δ-TOH according to the position and quantity of the methyl group in the chroman ring; α-TT, β -TT, γ-TT, δ-TT four kinds of tocotrienols. Among them, α-TOH has the highest biological activity, and the activities of γ-TOH and δ-TOH accou...

Claims

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Application Information

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IPC IPC(8): G01N24/08G01N33/50
CPCG01N24/088G01N33/5008G01N33/5044G01N33/5055
Inventor 耿越陆洋李慧
Owner SHANDONG NORMAL UNIV
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