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Mitochondrion correlation serum micro ribonucleic acid serving as markers of human obesity occurrence and application thereof

A marker, obesity technology, applied in the field of genetic engineering

Active Publication Date: 2015-09-30
NANJING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] At present, there is no report on the application of mitochondrial-associated miRNAs in the auxiliary diagnosis of obesity. If obesity-associated mitochondrial-associated miRNAs can be screened out as biomarkers and corresponding diagnostic kits can be developed, it will be a powerful impetus to the status quo of obesity diagnosis , and also opened up a new way for its drug screening, drug efficacy evaluation and targeted therapy

Method used

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  • Mitochondrion correlation serum micro ribonucleic acid serving as markers of human obesity occurrence and application thereof
  • Mitochondrion correlation serum micro ribonucleic acid serving as markers of human obesity occurrence and application thereof
  • Mitochondrion correlation serum micro ribonucleic acid serving as markers of human obesity occurrence and application thereof

Examples

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Embodiment 1

[0080] Example 1 Research Object Selection and Grouping Basis

[0081] From July 2013 to July 2014, the inventor collected obese cases and normal weight control adult blood samples from Jiangsu Provincial People's Hospital, the First Affiliated Hospital of Nanjing Medical University. The required 80 normal weight controls (average age: 22.15±3.45 years old) and 80 obese cases (average age: 22.32±3.51 years old) were used as experimental subjects for qRT-PCR detection of miRNA expression. Specific sample classification criteria are as follows:

[0082] (1) According to the Chinese reference standard: underweight (BMI<18.5), normal (BMI 18.5-23.9), overweight (BMI≥24), obese (BMI≥28); repeated BMI testing, confirmed obesity;

[0083] (2) Excluding those under the age of 18, athletes, those who are doing weight training, pregnant or breastfeeding, frail or sedentary;

[0084] (3) A normal weight (i.e. BMI 18.5-23.9) control matched with the age of the case;

[0085] (4) Grouping...

Embodiment 2

[0089] Example 2 Mitochondrial-related miRNA TaqMan array screening

[0090] Preparation of cDNA samples: a) Take 100 μl of serum; b) Add 900 μl of TRIzol, shake and mix, centrifuge at 12,000 rpm at 4°C for 15 minutes, and discard the waste liquid in the lower layer; c) Add 1.5 times the volume of supernatant, shake and mix, transfer to the spin column, centrifuge at 12000rpm for 15 seconds, and discard the lower layer of waste liquid; d) add 700 μl of RWT buffer to the spin column, centrifuge at 10000 rpm for 15 seconds, and discard the lower layer of waste liquid. e) Add 500 μl of RPE buffer to the spin column, centrifuge at 10,000 rpm for 15 seconds, and discard the lower layer. f) Repeat e. g) Add the spin column to a new 2ml tube and centrifuge at 10000rpm for 1 minute to remove the RPE buffer. h) Add 50 μl of DEPC-treated water to the column and centrifuge at 12000 rpm to collect RNA. i) Then cDNA is obtained by RNA reverse transcription reaction. The reverse transcr...

Embodiment 3

[0101] Example 3 qRT-PCR method to measure the expression of mitochondria-related miRNA in serum

[0102] Primers (Table 1) were designed for quantitative Real-time PCR detection of each miRNAs in the serum of 80 normal weight controls and 80 obese cases. Preparation of cDNA samples: a) Take 100 μl of serum; b) Add 900 μl of TRIzol, shake and mix, centrifuge at 12,000 rpm at 4°C for 15 minutes, and discard the waste liquid in the lower layer; c) Add 1.5 times the volume of supernatant, shake and mix, transfer to the spin column, centrifuge at 12000rpm for 15 seconds, and discard the lower layer of waste liquid; d) add 700 μl of RWT buffer to the spin column, centrifuge at 10000 rpm for 15 seconds, and discard the lower layer of waste liquid. e) Add 500 μl of RPE buffer to the spin column, centrifuge at 10,000 rpm for 15 seconds, and discard the lower layer. f) Repeat e. g) Add the spin column to a new 2ml tube and centrifuge at 10000rpm for 1 minute to remove the RPE buffer....

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Abstract

The invention belongs to the field of gene engineering, and discloses mitochondrion correlation serum micro ribonucleic acid serving as markers of human obesity occurrence and application thereof. The markers are some miRNA which is mitochondrion correlation miRNA and is selected from hsa-miR-141-3p, hsa-miR-196a-5p, hsa-miR-210-3p, hsa-miR-378a-3p, hsa-miR-484 and hsa-miR-499a-5p. Obesity cases and normal body weight can be well contrasted and separated, and the markers can be used for preparing obesity diagnosing or monitoring kits.

Description

field of invention [0001] The invention belongs to the field of genetic engineering, and relates to mitochondria-associated serum microRNA as a marker for human obesity and its application Background technique [0002] Obesity is a chronic metabolic disease characterized by energy metabolism imbalance, mainly manifested as excessive total fat content in the body and / or increased local content and abnormal distribution. It is a high-risk disease for the three major diseases of cardiovascular disease, diabetes and cancer. factor. Epidemiological surveys show that the incidence of obesity is increasing at an alarming rate, regardless of the well-off groups in economically developed countries or developing countries, which has constituted a serious problem in global medicine and public health in the 21st century. It is estimated that by 2030, 58% of adults will be obese and overweight. The incidence of obesity is increasing among adolescents and children, and its complications...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6883C12Q2600/158C12Q2600/178
Inventor 陆春城季娟秦玉峰郭锡熔季晨博夏彦恺王心如
Owner NANJING MEDICAL UNIV
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