Disease model animal expressing MEGSIN/RAGE/iNOS and method of evaluating compound with the use of the animal

A technology of disease models and compounds, applied in biochemical equipment and methods, chemical instruments and methods, disease diagnosis, etc., can solve problems such as unsatisfactory needs

Inactive Publication Date: 2007-02-14
RENASCI +1
View PDF2 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, megsin-Tg cannot meet the needs of animal models of diabetic complications mentioned above

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Disease model animal expressing MEGSIN/RAGE/iNOS and method of evaluating compound with the use of the animal
  • Disease model animal expressing MEGSIN/RAGE/iNOS and method of evaluating compound with the use of the animal
  • Disease model animal expressing MEGSIN/RAGE/iNOS and method of evaluating compound with the use of the animal

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0134] [Example 1] Preparation of megsin / RAGE / iNOS-Tg

[0135] RAGE / iNOS-Tg mice (Yamamoto, Y., etc., J.Clin.Invest., 108:261-268 (2001)) and megsin-Tg mice (Miyata, T., etc., J.Clin. Invest., 109:585-593 (2002)) to obtain megsin / RAGE / iNOS-Tg mice. Genomic DNA was extracted from the tail tissue, and as described in the literature (Yamamoto, Y., et al., J.Clin.Invest., 108:261-268 (2001); Miyata, T., et al., J.Clin.Invest., 109:585-593 (2002)) to confirm expression of RAGE, iNOS and megsin by PCR. Specifically, a part of the tail of Tg mice was excised at 4 weeks or later after birth, and genomic DNA was extracted using a DNA extraction kit (Qiagen Tissue Kit; Qiagen). Using this genomic DNA as a template, fragments of the introduced genes were amplified by PCR to confirm the presence of each introduced gene. For PCR detection of megsin, β-gl-3 primers (5'-CTT CTG GCG TGT GAC CGG CG-3' / SEQ ID NO: 4) and hM2-2 primers (5'-ATC GAA TTC TGA GAT CAT ( figure 1 ). For the deter...

Embodiment 2

[0136] [Example 2] Pathological examination results in megsin / RAGE / iNOS-Tg

[0137] Kidneys were excised from 16-week-old megsin / RAGE / iNOS-Tg mice, and PAS staining and PAM staining of glomeruli were performed according to conventional methods and the method described in WO 01 / 24628. Carnoy's fixative-fixed tissues were embedded in paraffin and sectioned at 4 microns for PAM staining. PAS staining (periodic acid Schiff reaction) generally stains polysaccharides red. In renal pathology, mesangial matrix hyperplasia and basement membrane thickening can be visualized by red staining of glycoproteins that are constituents of the mesangial matrix and basement membrane. Meanwhile, in PAM staining (periodic acid-hexamethylenetetramine silver salt staining method), components of the basement membrane and connective tissue are usually stained black (silver impregnation). In renal pathology, mesangial matrix sclerosis (fibrosis) and basement membrane thickening are stained black.

[...

Embodiment 3

[0140] [Example 3] Kidney weight, urine and blood biochemical tests in megsin / RAGE / iNOS-Tg

[0141] Kidney weight and urine and blood biochemical parameters were measured in 16-week-old megsin / RAGE / iNOS-Tg mice (Table 1). Other Tg mice (iNOS-Tg, RAGE / iNOS-Tg, megsin / iNOS-Tg, megsin / RAGE-Tg, megsin-Tg and RAGE-Tg) and wild-type mice (CD-1) were used as controls for comparison . The items to be measured are as follows: body weight (BW); kidney weight (KW); serum: total protein (TP: biuret method); triglyceride (TG: LPL / GK / GPDH-diaphorase / formazin staining method); total cholesterol (Tcho: enzymatic method); blood urea nitrogen (BUN: urease-ultraviolet light method); creatinine (Cr: alkaline picrate method); insulin (insulin: ELISA); blood glucose level (GLU: GOD / POD method); GOT; and GPT; urine: albumin (HbA1c: latex agglutination inhibition method); total protein (TP: pyrogallol red method); urea nitrogen (BUN: urease-ultraviolet light method); and creatinine (Cr: alkaline ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
molecular weightaaaaaaaaaa
Login to view more

Abstract

A triple Tg (megsin / RAGE / iNOS-Tg) is constructed by crossing a megsin-Tg with an RAGE / iNOS-Tg. It is found out that this megsin / RAGE / iNOS-Tg suffers from the onset of remarkable diabetic kidney lesion at an early stage which has never been observed so far and shows various uniform lesion images such as glomerular enlargement. It is also found out that the animal showing these symptoms is useful as a model animal of diabetic nephropathy. That is to say, the disease model animal as described above overexpresses megsin gene, receptor for advanced glycation end product gene and inducible nitric oxide synthase gene and, therefore, suffers from renal dysfunction associated with glomerular disorders at an early stage.

Description

technical field [0001] The present invention relates to a disease model animal, in particular, a model animal with renal dysfunction (for example, diabetic nephropathy) for hyperglycemia, and a model animal with renal dysfunction for glomerulopathy. The present invention also relates to a method for evaluating the curative effect of the compound on renal dysfunction by using the disease model animal. Background technique [0002] In recent years, due to the gradual westernization of lifestyle and diet, the number of diabetic patients has increased steadily. Diabetes is a disease in which persistent hyperglycemia causes various tissue disorders (called complications) throughout the body. In addition to the so-called macrovascular complications such as coronary arteriosclerosis and cerebrovascular disorders, pathological symptoms characteristic of diabetes arise: retinopathy, neuropathy and nephropathy, collectively referred to as "three major complications". In particular, ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): A01K67/027C12N15/09G01N33/50C07K14/47C12N9/02C12N15/85
CPCC12N9/0075C12N15/8509G01N2800/042G01N33/5088A01K2227/105A01K2217/05A01K2267/03A01K67/0275C07K14/47G01N2800/347A01K67/027C12N15/09
Inventor 宫田敏男黑川清山本博冈本宏
Owner RENASCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap