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Gene correlative to cardiovascular pathology caused by diabetes, and usage

A residue, myocardial technology, applied in the biological field

Inactive Publication Date: 2007-02-21
SHANGHAI INST FOR ENDOCRINE & METABOLIC DISEASES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, so far, little is known about the genes associated with diabetic cardiovascular disease. Therefore, there is an urgent need to find and develop new diabetic cardiovascular disease-related genes and their encoded proteins.

Method used

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  • Gene correlative to cardiovascular pathology caused by diabetes, and usage
  • Gene correlative to cardiovascular pathology caused by diabetes, and usage
  • Gene correlative to cardiovascular pathology caused by diabetes, and usage

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0119] Example 1. Preparation of diabetic rat model

[0120] 1.1 Animals and materials

[0121] 100 clean-grade male SD (Sprague-Dawley) rats, 2 months old, weighing (250±20) g, purchased from BK company, animal certificate number: Hudong 152; STZ (streptozotocin) purchase Provided from Sigma; blood glucose, triglyceride, cholesterol and other biochemical test kits were purchased from Mingdian Bioengineering Co., Ltd.; neutral monomodal pure insulin was purchased from Wanbang Biochemical Pharmaceutical Co., Ltd.; insulin primary antibody and related immunohistochemical reagents were purchased From Maixin Company; Rat-specific insulin radioimmunoassay kit was purchased from Linco Company.

[0122] Diet composition: average calories of ordinary feed is 3.9cal / g, of which carbohydrate accounts for 60%, protein accounts for 22%, fat accounts for 10% (mainly soybean oil), and other ingredients including cellulose account for 8%; average high-fat feed The calorie is 5.1cal / g, of which c...

Embodiment 2

[0143] Screening of genes related to early cardiomyopathy of type 2 diabetes in rats

[0144] 2.1 Materials

[0145] 2.1.1 Main equipment

[0146] Genomyx LRS fluorescence differential display analysis system (Beckman), the system includes (1) Genomyx LR electrophoresis instrument, including automatic drying gel device (2) Genomyx SC scanner (3) analysis software system including Acquire SC, Clarity SC, Extend LRS, Virtual Grid System, etc. (4) Glue cutting workstation. Spectrophotometer (Beckman DU650), 9700 PCR instrument (PE), Fluor-s TM Multilmager gel image analyzer (Bia-RAD), low temperature high speed centrifuge (Beckman), hybridization oven (Robbins Scientific 2000).

[0147] 2.1.2 Main reagents

[0148] HIEROGLPH mRNA Profile Kit (Beckman) includes: (1) 12 3'-end unlabeled fluorescent T7 (dT12) anchored primers (Anchored Primer, AP) (2) 20 5'M13r random primers (Arbitrary Primer, ARP) ). The Fluoro-DDRT-PCR kit (Beckman) includes: (1) 12 DNA molecular weight markers conta...

Embodiment 3

[0294] The expression of RDCR3 under high glucose culture condition is higher than that under low glucose culture condition

[0295] In this example, the primary cardiomyocytes of newborn rats were cultured and intervened in DMEM medium under conditions of high sugar (concentration 25 mmol / l) and low sugar (5.6 mmol / l). After 4 days of culture, the total mRNA of the cardiomyocytes was extracted, and the primers shown in SEQ ID NO: 5 and 6 were used to perform conventional RT-PCR to detect the expression of RDCR3.

[0296] The results showed that in the case of high glucose, the expression of RDCR3 gene was higher than that of low glucose (p<0.05)

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Abstract

This invention provides a new diabetic cardiovascular disease related gene, rat diabetic cardiomyopathy related gene 3 (RDCR3), polypeptides for coding RDCR3 protein, and method for producing RDCR3 protein by recombination technology. This invention also discloses the application of polypeptides coding RDCR3 protein. RDCR3 protein has a Laminin A domain, and is related to heart development and cardiomyopathy.

Description

Technical field [0001] The present invention belongs to the field of biotechnology. Specifically, the present invention relates to a new polynucleotide encoding rat diabetic cardiomyopathy related protein RDCR3 (Rat Diabetic Cardiomyopathy Related gene 3, rat diabetic cardiomyopathy related gene 3) , And the polypeptide encoded by this polynucleotide. The invention also relates to the use and preparation of the polynucleotide and polypeptide. Background technique [0002] Type 2 diabetic cardiovascular disease is not only the main cause of death for patients, but also has a higher incidence, rapid development process, and severe disease than the general population. It is necessary to strengthen the study of its pathogenesis. [0003] Although people have recognized the non-enzymatic glycation of proteins, excessive production of free radicals, hypermetabolism of polyols, polymorphisms of lipoproteins, cytokines and growth factors and the interaction of various mediators in the pr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/435C12N15/12C12N15/63C12P21/00A61K38/17A61P3/10A61P9/00G01N33/563
Inventor 罗敏
Owner SHANGHAI INST FOR ENDOCRINE & METABOLIC DISEASES