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MiRNA marker with lipid synthesis capability under whole grain diet

A marker, whole grain technology, applied in the direction of DNA/RNA fragment, recombinant DNA technology, microbial determination/inspection, etc., can solve the problem of lipid metabolism without biomarkers

Active Publication Date: 2020-12-22
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is currently no well-established evaluation system to assess the functional properties of whole grain diets, nor corresponding biomarkers to indicate the regulation of lipid metabolism by whole grain diets

Method used

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  • MiRNA marker with lipid synthesis capability under whole grain diet
  • MiRNA marker with lipid synthesis capability under whole grain diet
  • MiRNA marker with lipid synthesis capability under whole grain diet

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Example 1 Serum marker screening

[0067] (1) Selection of serum miRNA

[0068] The miRNAs with significant differences in the whole grain diet intervention were screened from the serum miRNA expression profile chip, and the miRNAs involved in lipid metabolism were analyzed through the miRNAs related database and online data analysis software, and the miRNA was screened out as miR-27a-3p.

[0069] (2) Extraction of serum miRNA

[0070] The collected plasma was allowed to stand at 4°C for 30 minutes, and then centrifuged at 4,000×rpm for 15 minutes at 4°C to obtain serum, which was stored at -80°C for later use. The miRNA in serum was extracted using Beyontian’s RNAeasy TM Animal RNA extraction kit (spin column type) for extraction, the specific steps are as follows:

[0071] a. Sample processing: 200 μl of lysate was added to 200 μl of serum for lysis.

[0072] b. Add an equal volume of binding solution to the centrifuge tube, and gently invert and mix 3-5 times. L...

Embodiment 2

[0099] Example 2 Detection of expression of proteins related to lipid synthesis in liver and intestine

[0100] The mice were fed under the intervention of three different dietary patterns (CS, BR, WW) for 8 weeks, and then serum and tissues were collected for index detection. Triglyceride and cholesterol kits from Japan WAKO Company were used to detect the contents of triglyceride and cholesterol in serum and liver.

[0101] (1) Extraction of liver and intestinal tissue proteins: Collect cells or a certain amount of mouse liver tissue and lyse or homogenize in RIPA lysate containing protease inhibitors and phosphatase inhibitors. Place on ice for 30 minutes, centrifuge at 14,000 rpm, 4°C for 20 minutes, collect the supernatant solution as the total protein of the tissue; RIPA lysate: 50mM Tris-HCl, pH=7.4; 150mM NaCl; 1mM EDTA; 1% Tritonx-100 ; Protease Inhibitor; Phosphatase Inhibitor.

[0102] (2) Sample preparation: After measuring the protein concentration with the BCA ...

Embodiment 3

[0127] Example 3 miR-27a-3p is used to detect and evaluate the lipid metabolism ability of mice at different time points

[0128] The mice were fed under the intervention of three different dietary patterns (CS, BR, WW) for 8 weeks.

[0129] (a) The expression level of miR-27a-3p, triglyceride and cholesterol levels in serum or plasma of samples were detected after 2 weeks, 4 weeks and 8 weeks of whole grain diet intervention.

[0130](b) According to the test results of step (a), judge the levels (contents) of triglyceride, cholesterol and miR-27a-3p in serum or plasma of the tested samples at different time points after eating the whole grain diet, and evaluate miR- Sensitivity of 27a-3p to lipid metabolism. The judging method was as follows: the expression level of miR-27a-3p in serum or plasma, as well as the levels of triglyceride and cholesterol were detected at 2 weeks, 4 weeks and 8 weeks of whole grain diet intervention, and compared with those of the control group (...

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Abstract

The invention discloses a miRNA marker with lipid synthesis capability under whole grain diet, and belongs to the field of biological medicine and food nutrition and health. The lipid metabolism of anorganism is intervened through the whole grain diet, and the biological action of miR-27a-3p in the lipid synthesis process is verified by performing lipid metabolism analysis on the organism, performing serum miRNAs sequencing analysis and evaluating the efficacy of miR-27a-3p in the lipid metabolism function. The miR-27a-3p can be used as a novel serum miRNA biomarker for regulating and controlling the body lipid metabolism by the whole grain diet.

Description

technical field [0001] The invention relates to a miRNA marker of lipid synthesis ability under whole grain diet, especially a serum or plasma miRNA marker of lipid synthesis ability under whole grain diet, belonging to the fields of biomedicine, food nutrition and health. Background technique [0002] For a long time, overweight and obesity, as a worldwide epidemic disease, has been a health issue of widespread concern, and in recent years has gradually shown a trend of rejuvenation—the prevalence of overweight or obesity in children and adolescents is increasing year by year. In addition, overweight and obesity can also cause a series of metabolic diseases including diabetes and cardiovascular disease, which seriously threaten human health. Due to its complex mechanism of action, controlling the balance of lipid metabolism in the body is very important for human health. [0003] Existing methods for detecting lipid metabolism in the human body are mainly to detect the blo...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12N15/113
CPCC12Q1/6888C12Q2600/178
Inventor 王立刘金鑫李言樊铭聪钱海峰
Owner JIANGNAN UNIV
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