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Molecular target for preparing medicine for diagnosing and treating type 1 diabetes and application of molecular target

A type 1 diabetes and molecular target technology, applied in the field of molecular targets, to achieve the effect of promoting β-cell apoptosis and inhibiting the proliferation of pancreatic β-cells

Active Publication Date: 2020-12-15
THE SECOND XIANGYA HOSPITAL OF CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The present invention provides a molecular target for preparing drugs for diagnosing and treating type 1 diabetes and its application, so as to solve the technical problems of preparing drugs for diagnosing and treating type 1 diabetes

Method used

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  • Molecular target for preparing medicine for diagnosing and treating type 1 diabetes and application of molecular target
  • Molecular target for preparing medicine for diagnosing and treating type 1 diabetes and application of molecular target
  • Molecular target for preparing medicine for diagnosing and treating type 1 diabetes and application of molecular target

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Expression of mIR-203a in normal mouse islet β cells and type 1 diabetes mouse islet β cells

[0034] Expression of mIR-203a in islet β cells

[0035] 1) Obtain the samples of pancreas tissues of type 1 diabetes (the tissue samples are normal mouse pancreas ①, type 1 diabetes mice insulitis stage ② and type 1 diabetes mice pancreas tissues of onset stage ③), and routinely dehydrate and soak the samples. Wax, embedding. The slice thickness is 6-8um.

[0036] 2) Paraffin sections were routinely dewaxed to water. Under the condition of 60°C, bake the slices for 30-60 minutes, put them in xylene while it is hot, let it stand for 10 minutes, transfer to another new xylene, let it stand for 10 minutes; then put it in 100% ethanol, 3 minutes, 95% ethanol 5 minutes, 75% ethanol for 5 minutes, 50% ethanol for 3 minutes, enzyme-free water for 3 minutes, and finally washed with PBS for 3 times, each time for 3 minutes. 3%H 2 o 2 , 5-10min at room temperature to ina...

Embodiment 2

[0055] Effects of mIR-203a on apoptosis of islet β cells

[0056] (1) MIN6 mouse islet β cell line was cultured in high-glucose DMEM medium (containing 15% FBS, 100 μg / mL streptomycin, 100 U / mL penicillin and 50 μM β-mercaptoethanol, and the glucose concentration was 4.5 g / L). The incubator conditions were set as: temperature 37°C, 5% CO2. The medium was replaced every 2-3 days, and when the cell density reached about 80% of the bottom area of ​​the culture flask, the culture medium was passaged at a ratio of 1:3.

[0057] (2) Perform miRNA transfection according to the instruction manual of GenePharma transfection reagent siRNA-mate (GenePharma, Shanghai, China):

[0058] 1) Plant MIN6 cells in well plates 18-24 hours before transfection, so that the density of transfection reaches about 50%;

[0059] 2) Add Opti-MEM (200 μL / well for 6-well plate, 50 μL / well for 24-well plate) into a sterile EP tube, then add miRNA mimics or inhibitors, siRNA (or corresponding NC) and mix g...

Embodiment 3

[0078] Therapeutic effect of mIR-203a inhibitor on diabetic mice

[0079] 1. NOD / ShiLtJ mice were cared for in accordance with the standard protocol of the Amgen Institutional Animal Care and Use Committee;

[0080] 2. When the blood glucose of NOD / ShiLtJ mice was 250-300mg / dL, and it was detected twice in 24 hours that it showed hyperglycemia. 2×107 mIR-203, mIR-203a inhibitor adenovirus (Sigma, St.Louis, MO, USA) was injected into the tail vein. The changes in blood sugar levels were detected, and the results showed that the mIR-203a inhibitor treatment group could reduce blood sugar levels and reverse the potential of diabetes onset.

[0081] Figure 9 It is the graph of the influence of mIR-203a on the blood sugar of diabetic mice, after the tail vein of the mIR-203a inhibitor adenovirus vector or the non-specific sequence negative control adenovirus vector (mIR-203a inhibitor NC) treated the newly-onset NOD / ShiLtJ, the Effects on blood sugar levels, the test results sh...

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Abstract

The present invention discloses a molecular target for preparing a medicine for diagnosing and treating diabetes and an application of the molecular target, the molecular target is mIR-203a and a basesequence of the mIR-203a is SEQ ID NO:1. The molecular target mIR-203a can accurately reflect a progress condition of islet function damages of the type 1 diabetes, and further can be used for diagnosing the diabetes. Meanwhile, the mIR-203a also has capabilities of obviously inhibiting proliferation of pancreatic beta cells and promoting apoptosis of the pancreatic beta cells, and can be used asthe molecular target for treating the type 1 diabetes. Therefore, the molecular target mIR-203a has important scientific research theory and clinical application value, provides a new clue and basisfor diagnosis, treatment or prognosis of the type 1 diabetes, and can be applied to preparing the medicine for diagnosing and treating the type 1 diabetes.

Description

technical field [0001] The invention relates to the field of molecular targets, in particular to a molecular target for preparing drugs for diagnosing and treating type 1 diabetes and its application. Background technique [0002] Type 1 diabetes is a common metabolic disease, and its morbidity and mortality have been gradually increasing in recent years. Since the evolution of type 1 diabetes is a relatively complex pathological process, which brings certain limitations to clinical diagnosis and treatment, it is urgent to provide a drug to provide new clues and basis for the diagnosis, treatment or prognosis of type 1 diabetes . Contents of the invention [0003] The invention provides a molecular target for preparing a drug for diagnosing and treating type 1 diabetes and its application, so as to solve the technical problem of preparing a drug for diagnosing and treating type 1 diabetes. [0004] The technical scheme that the present invention adopts is as follows: ...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12Q1/6883A61K45/00A61P3/10
CPCC12N15/113C12Q1/6883A61K45/00A61P3/10C12N2310/141C12Q2600/158C12Q2600/178
Inventor 周智广俞海波
Owner THE SECOND XIANGYA HOSPITAL OF CENT SOUTH UNIV
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