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Application of adipose-derived mesenchymal stem cell exosomes in preparation of medicine for treating metabolic syndrome

A metabolic syndrome and fat-derived technology, which is applied in the application field of fat-derived mesenchymal stem cell exosomes in the preparation of drugs for the treatment of metabolic syndrome, and achieves the effect of relieving insulin resistance and metabolic disorders, and the operation is simple and feasible

Inactive Publication Date: 2017-07-25
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The current research on adipose-derived mesenchymal stem cells mainly involves the use in kidney and fundus diseases (CN102048756A), the use in the treatment of erectile dysfunction (CN105902568A), the use in the preparation of liver fibrosis treatment preparations (CN103436555A), in the preparation of The use of preparations for the treatment of osteoarthritis (CN105748519A), the use in the preparation of chemotherapy sensitization preparations for liver cancer (CN103451152A), etc., but so far there is no information about the use of adipose-derived mesenchymal stem cells and their exosomes in the treatment of metabolic syndrome report

Method used

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  • Application of adipose-derived mesenchymal stem cell exosomes in preparation of medicine for treating metabolic syndrome
  • Application of adipose-derived mesenchymal stem cell exosomes in preparation of medicine for treating metabolic syndrome
  • Application of adipose-derived mesenchymal stem cell exosomes in preparation of medicine for treating metabolic syndrome

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Embodiment 1: the extraction and separation of ADSC

[0045] (1) Isolation of adipose tissue: select 10-12 weeks old C57BL / 6 male mice, anesthetize and kill the mice, open the abdominal cavity, peel off the fat in the epididymis, remove the blood vessels as much as possible, weigh the weight of the fat, and immerse it in PBS on ice placed on.

[0046] (2) Preparation of digestive juice: According to the weight of fat, 3ml of KRB buffer solution and 6mg of type Ⅰ collagenase are required for each gram of fat.

[0047] (3) Digestion of adipose tissue: Cut the stripped adipose tissue mechanically, add it to the prepared digestive solution, mix well, place it in a shaker at 37°C for 45 minutes, centrifuge at 2000rpm for 5 minutes, carefully remove the cells at the bottom and place Place on ice in KRB without enzyme.

[0048](4) Filtration: filter the cell suspension with a copper mesh with a pore size of 100 μm, centrifuge the filtered suspension at 2000 rpm for 5 minutes...

Embodiment 2

[0052] Example 2: Extraction and identification of ADSC-derived exosomes

[0053] ADSCs were cultured with exosome-free 10% fetal bovine serum DMEM medium (containing bFGF). Take the fourth-generation ADSC cell culture supernatant, filter it with a 0.22μm sterile filter, centrifuge at 5000g in a 100K concentrated centrifuge tube at 4°C for 30 minutes, take the concentrated solution from the upper chamber into a new sterile 15ml centrifuge tube, and add Exoquick-TC TM For the exosome extraction reagent, place at 4°C overnight, centrifuge at 5000g, 4°C for 30 minutes, discard the supernatant, resuspend the pellet with sterile PBS and aliquot, freeze at -80°C for storage.

[0054] The size and shape of the extracted exosomes were detected by transmission electron microscopy, and the results were as follows: figure 1 Shown; Exosome markers TSG101, CD9, CD63, and HSP90 were detected by western-blot method, and further identified, the results are as follows figure 2 shown.

Embodiment 3

[0055] Example 3: Effect of ADSC-derived exosomes on high-fat diet-induced metabolic syndrome model

[0056] (1) Establishment of high-fat diet-induced metabolic syndrome model and exosome intervention

[0057] Seven-week-old C57BL / 6 male mice were used to establish the metabolic syndrome model. After a week of pre-high-fat diet adaptation, they were fed a high-fat diet (HFD) (containing 60% cal), and fed a normal diet (NCD) as a control group. After 12-20 weeks of high-fat diet feeding, intraperitoneal injection of exosomes prepared in Example 2 (30 μg / mouse, injected once every 3 days, continuous injection for 6-8 weeks), namely the HFD-exosome group; intraperitoneal injection of normal saline The group is the control group, namely HFD-NS. NCD-NS was observed in mice fed normal diet and injected with normal saline intraperitoneally.

[0058] (2) Detection of triglyceride content in mouse serum

[0059] After the exosome intervention, the peripheral blood of the mice was ...

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Abstract

The invention discloses application of adipose-derived mesenchymal stem cell exosomes in preparation of a medicine for treating metabolic syndrome. The adipose-derived mesenchymal stem cell exosomes are prepared through the following method: (1) dispersing a stripped adipose tissue into a digestion solution to produce cell suspension; (2) centrifugating the cell suspension to form cell sediments, and separating and washing the sediments, thus obtaining an adipose-derived mesenchymal stem cell-containing blood vessel matrix component; (3) performing multiplication culture on ADSC (adipose-derived stem cell) in an in-vitro culture system, collecting supernate obtained by the multiplication culture for concentration, and adding an exosome extraction reagent into the concentrated supernate, thus obtaining the adipose-derived mesenchymal stem cell exosomes through extracting. The adipose-derived mesenchymal stem cell exosomes can effectively relieve insulin resistance and metabolic disorders and inhibit occurrence and development of the metabolic syndrome and non-alcoholic fatty liver disease to a certain extent.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to the application of adipose-derived mesenchymal stem cell exosomes in the preparation of drugs for treating metabolic syndrome. Background technique [0002] Metabolic syndrome refers to the pathological state of metabolic disorders of proteins, fats, carbohydrates and other substances in the human body. It is a group of complex metabolic disorder syndromes and is a risk factor for diabetes and cardiovascular and cerebrovascular diseases. Insulin resistance and a series of metabolic syndromes such as hyperglycemia, hypertension, and dyslipidemia are the "common soil" for the onset of non-alcoholic fatty liver disease, diabetes, and atherosclerosis. The high hospitalization rate, high disability rate, and high fatality rate caused by metabolic syndrome have become a public health problem that threatens the health of all mankind. [0003] Adipose-derived mesenchymal stem cells ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/28A61P3/00A61P1/16C12N5/0775
CPCA61K35/28C12N5/0667C12N2509/00
Inventor 王群赵汇张利宁朱法良郭春张慧颖
Owner SHANDONG UNIV
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