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

A technology for retinoic acid receptors and chronic kidney disease, applied in biological testing, biomaterial analysis, disease diagnosis, etc., can solve the problems of lack of sensitivity, specificity, and poor specificity in risk stratification, and achieve the purpose of guiding clinical treatment and judging prognosis , high specificity, good clinical treatment and prognosis

Active Publication Date: 2022-04-15
ZHONGSHAN HOSPITAL XIAMEN UNIV
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  • Summary
  • 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 a biomarker in chronic kidney disease
  • Application of retinoic acid receptor reactive protein 1 as a biomarker in chronic kidney disease
  • Application of retinoic acid receptor reactive protein 1 as a biomarker in chronic kidney disease

Examples

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 ( like 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 and ...

Embodiment 3

[0042] The human RARRES1 genome sequence was amplified from the SPORT 6 plasmid by PCR method, and a Xbal I 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 connecte...

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Abstract

The invention discloses the application of retinoic acid receptor reactive protein 1 as a biomarker in chronic kidney disease, belonging to the technical field of biology and medicine. The invention detects the expression level of retinoic acid receptor reactive protein 1 in the body fluid of patients with chronic kidney disease Changes in DN to predict the prognosis of nephropathy. For the first time in the present invention, retinoic acid receptor reactive protein 1 is used as a marker for chronic kidney disease, especially diabetic nephropathy. According to the evidence, it has important guiding significance for the clinical prognosis of chronic kidney disease, especially diabetic nephropathy, and helps to further understand the 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 Patents(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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