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Application of mesenchymal stem cell exosome in preparation of drugs for preventing or treating type 1 diabetes and related diseases thereof

A technology of type 1 diabetes and exosomes, applied in the field of biomedicine, to achieve the effect of reducing blood sugar rise, low toxicity, and preventing the onset of diabetes

Active Publication Date: 2021-12-07
SHANDONG UNIV QILU HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, in the prior art, there are no related reports that relevant miRNAs can be applied to the prevention or treatment of type 1 diabetes and its related diseases.

Method used

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  • Application of mesenchymal stem cell exosome in preparation of drugs for preventing or treating type 1 diabetes and related diseases thereof
  • Application of mesenchymal stem cell exosome in preparation of drugs for preventing or treating type 1 diabetes and related diseases thereof
  • Application of mesenchymal stem cell exosome in preparation of drugs for preventing or treating type 1 diabetes and related diseases thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] The extraction and identification of embodiment 1 mouse BM-MSC

[0040] (1) Separation of femur: select C57BL / 6 mice aged about 4 weeks, soak them in 75% ethanol for 5 minutes after overdose of anesthesia, and then move them into the ultra-clean bench to separate the bilateral femurs and femurs of the mice under aseptic conditions. tibia.

[0041] (2) Obtaining bone marrow: cut off both ends of the femur, expose the bone marrow cavity, use a 5mL syringe to draw complete medium (79% DMEM / F12 medium + 20% FBS + 1% double antibody) to wash the cavity, and repeatedly beat for 2- Bone marrow was harvested 3 times.

[0042] (3) Seeding plate: The bone marrow fluid extracted from one femur was inoculated into one well of a six-well plate for seeding, with about 2.5 mL per well. Put to 37℃, 5% CO 2 Cells were cultured overnight in a cell culture incubator, and replaced with new complete medium after 24 hours. In the subsequent culture, 10% FBS + 1% double antibody DMEM / F12 ...

Embodiment 2

[0044] Example 2 Extraction and identification of BM-MSC exosomes

[0045] After the P5 generation of BM-MSC was cultured, the cell culture supernatant (using fetal bovine serum without exosomes) was collected, and the exosomes in the medium were extracted by ultracentrifugation.

[0046]The specific operation in this example is as follows: take the CM blood culture supernatant, put it into a 50 mL centrifuge tube, and centrifuge at 3000 g for 30 min. Take the supernatant and put it into a high-speed centrifuge tube, centrifuge at 10000g for 30min to obtain the supernatant. Filter the supernatant with a 0.22 μm filter. The filtrate was put into a 38.5mL ultracentrifuge tube and centrifuged at 110000g for 70min. After carefully discarding the supernatant, the BM-MSC exosomes were obtained and resuspended in PBS, saline or liquid medium for use.

[0047] Observation of exosome morphology by (1) electron microscope, the observation results are as follows: image 3 shown. (2)...

Embodiment 3

[0049] Example 3 Effect of BM-MSC exosomes on the onset of diabetes in NOD mice

[0050] BM-MSC exosomes were injected into the tail vein of 3-week-old female NOD mice (150 μg / mouse), and the mice were injected once at the age of 5 weeks, 7 weeks and 9 weeks, and the dose was the same as before , carry out the following experiments respectively:

[0051] (1) Monitor blood sugar: take 2 consecutive random blood sugars ≥ 250 mg / dl as the standard for the onset of diabetes in NOD mice, and observe the onset of the mice. The statistical results of the onset of the mice are as follows: Figure 6 As shown, it can be seen from the figure that the control group without BM-MSC exosomes had no normal blood sugar mice at 30 weeks, while 80% of the mice in the experimental group added with BM-MSC exosomes still maintained normal blood sugar without disease Therefore, it can be concluded that BM-MSC exosomes can significantly reduce the incidence of NOD mice.

[0052] (2) Serological ind...

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Abstract

The invention provides an application of miRNA (micro Ribonucleic Acid) in drugs for treating type 1 diabetes and related diseases thereof. The application is characterized in that the miRNA is miR-199a-5p. The new application of the miR-199a-5p in preparation of drugs for treating type 1 diabetes and related diseases thereof and the new application of the miR-199a-5p in preparation of preparations for inducing T cell differentiation are provided for the first time; the obtained drugs can effectively reduce blood sugar rise caused by the type 1 diabetes, improves islet inflammation to a certain extent, and achieves the effect of preventing and treating the type 1 diabetes; and the miR-199a-5p is adopted as an effective component, so that the miR-199a-5p has high drug targeting property and has the advantages of trace amount, high efficiency, safety, reliability and low toxicity.

Description

technical field [0001] This application relates to the field of biomedical technology, in particular to the application of miR-199a-5p in the preparation of drugs for preventing or treating type 1 diabetes and related diseases. Background technique [0002] Type 1 diabetes mellitus (T1DM) is an autoimmune disease, often with acute onset, often accompanied by severe complications such as ketoacidosis. [0003] The pathogenesis of T1DM is mainly the abnormal proliferation and activation of reactive T cells in the body. The pro-inflammatory cells Th1 and Th17 cells proliferate, while the anti-inflammatory regulatory T cells (Treg) and Th2 cells are relatively insufficient, resulting in Th1 / Th2 And Th17 / Treg cell ratio imbalance, the formation of peripancreatic inflammation, and then cause the destruction of islet β cells, insulin secretion decreased, blood sugar increased. Therefore, effectively inhibiting the proliferation of pro-inflammatory Th1 and Th17 cells in NOD mice an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/7088A61K35/28A61P3/10
CPCA61K31/7088A61K35/28A61P3/10
Inventor 段武
Owner SHANDONG UNIV QILU HOSPITAL
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