A kind of hleg1 gene and its application and medicine

A gene and drug technology, applied in the hLeg1 gene and its application and drug fields, can solve the problem of few drug targets, and achieve the effect of promoting fat synthesis and reducing fat accumulation

Active Publication Date: 2018-05-04
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, at present, there are still relatively few research results on drugs for the effective treatment of obesity, and there are relatively few drug targets related to obesity

Method used

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  • A kind of hleg1 gene and its application and medicine
  • A kind of hleg1 gene and its application and medicine
  • A kind of hleg1 gene and its application and medicine

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] 1. Northern blot analysis of the expression of mLeg1 in different tissues

[0049] The 8-week-old male mice with C57BL / 6 background were used as research objects, and the expression profile of mLeg1 gene was detected by Northern blot analysis. Using the antisense strand of the mLeg1 gene as a probe, Northern blot analysis was performed to analyze a series of digestive organs (heart, liver, pancreas, lung) of mLeg1 gene mice including the liver. , kidney (kidney), stomach (stomach), intestine (gut), salivary gland (SG)).

[0050] The experimental method of Northern blot analysis is as follows.

[0051] 1.1 RNA extraction:

[0052] 1.1.1 Take the tissue that needs to extract RNA, and grind it with liquid nitrogen until there are no obvious particles. During the grinding process, keep liquid nitrogen to prevent RNA degradation.

[0053] 1.1.2 Take 50-100mg sample and add 1ml Trizol ( Reagent, Life Technologies, Cat.no.15596-026), fully homogenized by pumping with a 26...

Embodiment 2

[0075] Since the homolog protein Leg1 of mLeg1 protein in zebrafish is a secreted protein, the above results have shown that mLeg1 gene is mainly expressed in salivary glands, but its mLeg1 protein may also be secreted and transported to other tissues to play a role. Therefore, the inventors of the present invention extracted the total protein from different tissues of mice, and detected the distribution of mLeg1 protein in different tissues by Western Blot.

[0076] Western Blot was used to detect the distribution of mLeg1 protein in different tissues. The experimental method is as follows.

[0077] 2.1 Protein extraction:

[0078] After the mice died suddenly, the target tissues (SG, liver, gut, blood, lung, heat, stomach, kindney, pancrease) were removed, placed in 1.5ml centrifuge tubes, and quickly frozen in liquid nitrogen to prevent degradation. When extracting protein, take out the sample, grind and pulverize by liquid nitrogen, collect the sample powder in a centrif...

Embodiment 3

[0086] Since the salivary gland is a secretory gland, its most important function is to secrete saliva, and mLeg1 is also a secretory protein. Therefore, the inventors of the present invention investigated whether mLeg1 is secreted into saliva.

[0087] The content of mLeg1 in saliva was detected by Western Blot. Specific steps are as follows.

[0088] 3.1 Saliva collection: 0.5 mg / kg of Pilocarpine (Sigma) was injected intraperitoneally into the mice, and a capillary was placed in the mouth of the mice to drain and collect the secreted saliva. Pilocarpine, a drug used to treat dry mouth, stimulates a flood of saliva. The saliva secreted by wild-type mice and mLeg1 systemic knockout mice (how to obtain them are described below) were collected respectively.

[0089] 3.2 Saliva treatment: the collected saliva was boiled at 100° C. for 5 minutes with 5x Laemmli buffer (10% SDS, 250 mM Tris-HCl, 0.1‰ Bromphenol blue, 500 mM DTT, 50% Glycerol) of 1 / 5 saliva volume.

[0090] 3.3...

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Abstract

The invention provides an hLeg1 gene and the application and a drug thereof, and relates to the technical field of gene functions and application. In the invention, mLeg1 gene knockout mice are used as study subjects, a very comprehensive study is performed on the function of a mLeg1 gene by using means of genetics, molecular biology, biological chemistry and cytobiology. Study results show that mLeg1 protein can adjust and control signals of Akt in a body by EGFRs and further adjust and control fat synthesis in the body; the study results provide new means and ideas for treating obesity and recovering physique of cancer patients after chemotherapy through man-made intervention of expression levels of the hLeg1 gene and the hLeg1 protein at later period.

Description

technical field [0001] The present invention relates to the technical field of gene function and application, in particular, relates to a hLeg1 gene and its application and medicine. Background technique [0002] Over the past few years, obesity cases have grown rapidly worldwide and are now the fifth leading threat to human death. In developed countries, obesity has emerged as early as the 1980s, and cases have continued to increase since then, but the rate of increase has slowed down in the past 8 years; while in developing countries, obesity is increasing at a very rapid rate every year. speed is increasing. Although obesity generally does not lead directly to death, its complications, especially cardiovascular disease, can be fatal. In 2010, obesity was responsible for an estimated 3.4 million deaths. In addition, studies of obese patients in the United States have shown that obesity is likely to reduce the average life expectancy of humans in the future. According t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/12C07K14/47A61K45/00A61K48/00A61P3/04A61P3/10
CPCA61K45/00A61K48/005C07K14/4702
Inventor 彭金荣胡敏杰
Owner ZHEJIANG UNIV
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