Application of brown adipocyte-derived exosome

A brown fat and exosome technology, applied in the application field of brown fat cell-derived exosomes, can solve the problems of limited energy consumption, low adipose tissue survival rate, technical difficulty, etc.

Inactive Publication Date: 2021-07-06
JIANGSU PROVINCIAL HOSPITAL OF TCM
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the human body, brown adipose tissue and white adipose tissue are in a mixed state, which mainly exists near the scapula, around the kidneys and the spine, and does not have obvious distinguishable tissue blocks like rodents, and at the same time The survival rate of transplanted adipose tissue is low, so it is very technically difficult to treat fatty liver and other metabolic diseases by transplanting brown adipose tissue in humans
At the same time, the use of cold and other stimuli to activate human brown adipose tissue to produce heat and consume energy for treatment also has a large degree of limitation, because some obese people do not exist or only have a small amount of brown fat, so it consumes energy. limited capacity
However, there is no report on the treatment of fatty liver with exosomes derived from brown adipocytes.

Method used

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  • Application of brown adipocyte-derived exosome
  • Application of brown adipocyte-derived exosome
  • Application of brown adipocyte-derived exosome

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Isolation and culture of exosomes secreted by brown adipocytes.

[0026] (1) Extraction of exosomes from brown adipocytes

[0027] After the mice were killed by neck dislocation, the brown fat cells were removed from the mouse scapula, and a small amount of white fat above the brown fat was removed. Mince the brown adipocytes with scissors. After shredding, place them in DMEM containing 10% mv free FBS (both purchased from Gibco Company) culture medium, and then place them in a carbon dioxide incubator for cultivation. After culturing for 24 hours, the culture medium was collected for exosome extraction.

[0028] (2) Extraction of exosomes

[0029] First, pretreat the culture medium. The pretreatment method is 300xg, centrifuge for 10 minutes, remove the precipitate, 3000xg, centrifuge for 30 minutes, then remove the bottom, and finally 10000xg, centrifuge for 60 minutes, and then remove the precipitate. Pretreatment is to remove cell debris or some larger...

Embodiment 2

[0030] Example 2 Identification of brown adipocyte-derived exosomes

[0031] (1) Western-blot identification of exosome marker proteins CD63, CD9 and TSG101

[0032] Add appropriate amount of cell lysate. The cell lysate (purchased from Biyuntian) was lysed on ice for 30 minutes to allow the cell lysate to fully function. The cells in the cell lysate are ultrasonically disrupted to release the protein in the cells more completely. Centrifuge at 12000xg for 5 minutes at 4°C. Aspirate the supernatant and test the protein concentration. The protein concentration test method is the BCA method (purchased from Thermo Fisher), and the protein is diluted 10 times for measurement, that is, 1 μl of protein solution is added to 9 μl of ddH2O (see the relevant instructions for detailed steps). Adjust the egg whites to the same concentration, add 5x loading buffer (purchased from Beyond), and cook in a metal bath at 100°C for 5 minutes, then store the boiled egg whites at -20°C. Confi...

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PUM

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Abstract

The invention discloses an application of a brown adipocyte-derived exosome. The invention discloses application of the brown adipocyte-derived exosome in preparation of a medicine for treating non-alcoholic fatty liver, and preferably application in preparation of a medicine for treating non-alcoholic fatty liver caused by high fat obesity. The brown adipocyte-derived exosome is used for expressing symbolic proteins CD63, CD9 and TSG101. The extracted brown adipocyte-derived exosome can effectively reduce the weight of non-alcoholic fatty liver of high-fat-induced obese mice and the ratio of liver to volume, reduce the content of fatty acid and cholesterol in the liver, reduce the liver, slow down fat synthesis in the liver and increase the consumption of fatty acid at the same time. Therefore the invention provides a new way for treating non-alcoholic fatty liver of obese mice caused by high fat.

Description

technical field [0001] The present invention relates to the application of exosomes derived from brown adipocytes, in particular to the application of exosomes produced by brown adipocytes in the treatment of non-alcoholic fatty liver caused by obesity. Background technique [0002] Non-alcoholic fatty liver disease (NAFLD) is a common liver disease defined as fat accumulation in more than 5% of liver cells in the liver with no or light alcohol consumption. According to incomplete statistics, 14% to 27% of people worldwide suffer from fatty liver. The current drug treatment for fatty liver does not meet the clinical needs, and so far, there is no FDA-approved drug for the treatment of more severe non-alcoholic fatty liver. [0003] Brown adipose tissue is a kind of energy-producing tissue, which can convert the chemical energy stored in the body into heat energy to maintain body temperature through the uncoupling protein UCP1 in mitochondria, that is, to produce non-shock h...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/35A61P1/16
CPCA61K35/35A61P1/16
Inventor 郑莎莎方祝元
Owner JIANGSU PROVINCIAL HOSPITAL OF TCM
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