Application of retinoic acid receptor reactive protein 1 as biomarker in chronic kidney disease

A retinoic acid receptor and biomarker technology is applied in the application field of retinoic acid receptor reactive protein 1 as a biomarker in chronic kidney disease, which can solve the problems of lack of sensitivity, specificity and poor specificity of risk stratification. , to achieve the effect of guiding clinical treatment and judging prognosis, high specificity, good clinical treatment and judging prognosis

Active Publication Date: 2020-05-08
ZHONGSHAN HOSPITAL XIAMEN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Current prognostic studies mainly focus on changes in urinary protein and creatinine or glomerular filtration rate, but changes in urinary protein lack sensitivity and specificity for risk stratification of DKD progression, while changes in creatinine or glomerular filtration rate Susceptible to excessive diuresis in diabetic patients, nephrotoxic drug use, infection, etc., sometimes reflecting only transient changes in renal perfusion
For this reason, blood uric acid, cystatin C, inflammatory markers, cytokines (such as CTGF, FGF-23, sCD, etc.) 28 ) and markers such as miRNA, however, these markers still have disadvantages such as poor specificity;

Method used

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  • Application of retinoic acid receptor reactive protein 1 as biomarker in chronic kidney disease
  • Application of retinoic acid receptor reactive protein 1 as biomarker in chronic kidney disease
  • Application of retinoic acid receptor reactive protein 1 as biomarker in chronic kidney disease

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Experimental program
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Effect test

Embodiment 1

[0035] Upregulation of retinoic acid receptor reactive protein 1 in chronic kidney disease

[0036] Immunohistochemical staining of kidney biopsy specimens in the early stage of CKD (chronic kidney disease),

[0037] Collect kidney biopsy specimens of different types of CKD early stage (GFR>60ml / min) (FSGS, DKD, MCD, etc.) with complete clinical data (including etiology, ACR, GFR, CRP, etc.), and use the relatively normal specimens around the nephrectomy tumor The tissue was used as a control, and the paraffin section was subjected to antigen retrieval according to the conventional method, followed by immunohistochemical staining, using the Santa Cruz TIG1 antibody (sc-98965, diluted 1:50) as the primary antibody, horseradish peroxidase-labeled goat antibody Rabbit secondary antibody was used to detect the expression level of RARRES1. The result is as figure 1 as shown, figure 1 It revealed that the expression of RARRES1 was significantly up-regulated in the FSGS group and ...

Embodiment 2

[0039] The transmembrane protein RARRES1 can be cleaved to have a soluble splice body

[0040] It is generally accepted that RARRES1 is a type I transmembrane protein. In this example, the human RARRES1 subtype eukaryotic expression plasmid (pN1-RARRES1-v5) was transfected into 293T cells, and the cell membrane proteins were extracted using the instructions of the Biovision Cell Membrane Protein Extraction Kit (Catalog #K268), and loaded separately 30 μl of cytoplasmic protein, total cell membrane protein, and total cell protein were subjected to western-blot, and V5 antibody was used to detect RARRES1, and Pan-cadherin was used to detect known membrane proteins. The results suggested that membrane proteins were rich in RARRES1, which was consistent with the prediction of transmembrane proteins ( Such as figure 2 shown). At the same time, 24 hours after plasmid transfection, serum-free medium was used, cell culture supernatant was collected, and 30ug of cell total protein a...

Embodiment 3

[0042] The human RARRES1 genome sequence was amplified from the SPORT 6 plasmid by PCR method, and a XbalI restriction site was added at the C-terminus while carrying the V5 tag. The PCR product was then ligated into pGEMTasy vector (Promega A1360). At the same time, the human RARRES1 gene fragment was cloned into the pTRETight vector (Clontech: CAT NO.631059) with EcoR I and NheI restriction sites to make it contain AFEI restriction sites. Subsequently, the human RARRES1-V5 fragment was cloned into the above-mentioned vector digested with AFEI and NotI double enzymes, and the pTRE-hRARRES1-v5 vector was constructed. After the pTRE-hRARRES1-v5 vector was digested with EcoRI and NotI, it was inserted into the vector of pN1-EGFP digested by the same double digestion, and pN1-RARRES1-v5 was constructed ( Figure 5 ).

[0043] The human RARRES1 genome sequence was amplified from the SPORT 6 plasmid by PCR, StuI and XbaI restriction sites were designed at both ends, and connected...

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Abstract

The invention discloses an application of retinoic acid receptor reactive protein 1 as a biomarker in a chronic kidney disease. The invention belongs to the technical field of biology and medicine. According to the invention, the change of the expression level of retinoic acid receptor reactive protein 1 in the body fluid of a chronic kidney disease patient is detected, and prediction of prognosisof a kidney disease is carried out. According to the invention, the retinoic acid receptor reactive protein 1 is used as a marker of the chronic kidney disease, especially diabetic nephropathy for the first time, is high in specificity, can also provide a theoretical basis for formulation of a new DN pathological typing standard, has an important guiding significance for clinical prognosis of chronic kidney diseases, especially diabetic nephropathy, is beneficial to further understanding of pathogenesis of DN, and can better guide clinical treatment and judge prognosis.

Description

technical field [0001] The invention relates to the technical fields of biology and medicine, in particular to the application of retinoic acid receptor reactive protein 1 as a biomarker in chronic kidney disease. Background technique [0002] Diabetic nephropathy (DN, also known as DKD) is increasing worldwide, and DKD patients are at higher risk of end-stage renal disease (ESRD) and are the leading cause of dialysis in patients with chronic kidney disease (CKD). [0003] Therefore, early identification of high-risk groups that may develop ESRD is very important. Current prognostic studies mainly focus on changes in urinary protein and creatinine or glomerular filtration rate, but changes in urinary protein lack sensitivity and specificity for risk stratification of DKD progression, while changes in creatinine or glomerular filtration rate Susceptible to excessive diuresis in diabetics, nephrotoxic drug use, infection, and other factors, sometimes reflecting only transient...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/6893G01N2800/347G01N2800/52
Inventor 陈安群关天俊何慈江
Owner ZHONGSHAN HOSPITAL XIAMEN UNIV
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