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Application of Tumor Necrosis Factor α-Inducible Protein 3 in the Preparation of Drugs for Treating Diabetic Cardiomyopathy

A technology of tumor necrosis factor and diabetic cardiomyopathy, applied in the field of medicine, can solve the problems such as A20, which has not yet been seen, and achieve the effect of inhibiting myocardial hypertrophy, improving heart function, and high safety

Active Publication Date: 2020-07-10
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But so far, there is no relevant report on the role of A20 in diabetic cardiomyopathy.

Method used

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  • Application of Tumor Necrosis Factor α-Inducible Protein 3 in the Preparation of Drugs for Treating Diabetic Cardiomyopathy
  • Application of Tumor Necrosis Factor α-Inducible Protein 3 in the Preparation of Drugs for Treating Diabetic Cardiomyopathy
  • Application of Tumor Necrosis Factor α-Inducible Protein 3 in the Preparation of Drugs for Treating Diabetic Cardiomyopathy

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] [Example 1] Detecting the change of A20 protein level in the case of high-fat feeding

[0069] 1. Establishment of high-fat feeding-induced diabetic cardiomyopathy model

[0070] (1) Grouping of experimental animals:

[0071] Eight-week-old, male, C57 mice were selected and fed with two special diets, D12942 high-fat diet (High fatdiet, HFD) and D12450B normal diet (Normal chow, NC).

[0072] (2) Mouse heart sampling:

[0073] After 24 weeks of feeding, the hearts of mice fed HFD and NC diets were taken out, weighed, and stored in a -80°C refrigerator in liquid nitrogen for molecular biology experiments.

[0074] 2. Western blotting (WB) detection of A20 protein levels in the hearts of two groups of mice

[0075] 2.1 Extract protein samples:

[0076] (1) Autoclave the protein extraction equipment including ophthalmic scissors, ophthalmic straight tweezers, and steel balls.

[0077] (2) Quickly take out the required protein sample and place it on dry ice. Mark the ...

Embodiment 2

[0098] [Example 2] Effect of adenovirus-mediated A20 overexpression (AdA20) on the toxicity of primary cardiomyocytes stimulated by high levels of fatty acids

[0099] 2.1. Primary neonatal SD rat cardiomyocyte culture

[0100] (1) Eight newborn Sprague-Dawley suckling mice were disinfected with 75% alcohol below the neck, and the heart was removed with ophthalmic scissors and micro forceps, and placed in a glass plate filled with 10mL DMEM / F12 solution. Take another one and repeat the above process.

[0101] (2) Wash the heart with DMEM / F12 medium, and cut the heart into 1-2mm pieces. Transfer to a serum bottle with a rotor, suck off DMEM / F12, and add trypsin digestion solution. Rotate at 120r / min, digest for 15min, rest for a few seconds, and discard the supernatant.

[0102] (3) Add trypsin digestion solution, the rotation speed is 120r / min, and digest for 15min. Stand still for a few seconds, draw the supernatant, terminate the digestion with DMEM / F12 medium with 20% c...

Embodiment 3

[0119] [Example 3] A20 gene knockout promotes the occurrence and development of diabetic cardiomyopathy in mice

[0120] 1. Animal grouping:

[0121] 8-week-old, male, WT mice and A20-CKO mice were selected and fed with two special diets, D12942 high-fat diet (High fat diet, HFD) and D12450B normal diet (Normal chow, NC), respectively, namely WT NC group, CKO NC group, WT HFD group, and CKO HFD group. There are 4 groups in total. At the end of the 24th week, the hearts of the mice were taken out, and the hearts of the mice were taken out to take pictures, and then fixed in formaldehyde solution or frozen at -80°C for pathological analysis. For molecular biology testing.

[0122] 2. Materials:

[0123] (1) Preliminary work:

[0124] A urine cup containing 20 mL of 10% formaldehyde by volume was prepared in advance and labeled (mouse number, group, type of operation and date of collection). Place a petri dish filled with 10% KCl solution in mass fraction at the sampling site...

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Abstract

The invention discloses application of tumor necrosis factor alpha induced protein 3 in the preparation of medicine for curing diabetic cardiomyopathy. A mouse diabetic cardiomyopathy model is established through high fat diet (HFD) induction, primary rat myocardial cells are infected by constructing overexpression A20 recombinant adenovirus vector (AdA20), an in vitro cell model of diabetic cardiomyopathy is established by using high-level fatty acid stimulation, simultaneously WT mouse and heart specific A20 gene knockout mouse (A20-CKO) are taken as experimental subjects, the mouse diabeticcardiomyopathy model is established by using HFD induction, and the protein expression level of A20 inside the mouse heart tissue of HFD 24W is obviously reduced; and in the high fat diet induced mouse diabetic cardiomyopathy model, A20 has the effects of inhibiting cardiac hypertrophy, improving heart function and blocking the development of diabetic cardiomyopathy. Due to the fact that A20 is organism endogenous protein, the safety of the A20 as medicine is extremely high.

Description

technical field [0001] The invention relates to the technical field of medicine, in particular to the application of tumor necrosis factor alpha-induced protein 3 (TNFAIP3, also known as A20) in the preparation of drugs for treating diabetic cardiomyopathy. Background technique [0002] With the development of society, cardiovascular disease has become a huge problem faced by both developing and developed countries [1]. What needs special attention is that the sharp increase in the number of people with diabetes is one of the important reasons for the high incidence of cardiovascular diseases [2]. Diabetic cardiomyopathy is considered to be a change in myocardial structure and function caused by diabetes, while excluding ischemic diseases, hypertension, and other diseases that may cause cardiomyopathy [2-3]. The pathological features of diabetic cardiomyopathy are: hypertrophy of cardiomyocytes, interstitial fibrosis, infiltration of PAS-positive substances, thickening of t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/43A61K45/00A61K48/00A61P3/10A61P9/00C12Q1/6883
CPCA61K38/43A61K45/00A61K48/005C12Q1/6883C12Q2600/158
Inventor 李红良
Owner WUHAN UNIV
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