Application of long-chain non-coding RNA as biomarker to diagnosis of contrast-induced acute kidney injury (CI-AKI)

A long-chain non-coding, acute kidney injury technology, applied in the field of biomedicine, can solve the problems of poor specificity of serum creatinine, lack of targeted drugs, missing, etc., and achieve high fluid stability, high prediction efficiency, and high sequence homology Effect

Active Publication Date: 2021-01-15
GUANGDONG GENERAL HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The specificity of serum creatinine in this definition is poor, because of the fluctuation of serum creatinine due to the transfer of receptor fluid and the effect of drugs
In addition, the insensitivity and lateness of serum creatinine and the lack of accurate new biomarkers also bring great challenges to the early detection of CI-AKI.
In addition, since the key molecular mechanism and regulatory network of CI-AKI are still unclear, the effectiveness of classic prevention strategies such as applying saline hydration is controversial and limited, and there is a lack of specific targeted drugs.

Method used

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  • Application of long-chain non-coding RNA as biomarker to diagnosis of contrast-induced acute kidney injury (CI-AKI)
  • Application of long-chain non-coding RNA as biomarker to diagnosis of contrast-induced acute kidney injury (CI-AKI)
  • Application of long-chain non-coding RNA as biomarker to diagnosis of contrast-induced acute kidney injury (CI-AKI)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1, establish novel CI-AKI rat model

[0027] (1) 20 rats were randomly divided into the CI-AKI group and the control group, and were dehydrated for 24 hours. Six hours before exposure to the contrast agent, the rats in the two groups were injected with furosemide 15ml / kg intraperitoneally; Iopromide, a hypotonic contrast agent, was administered at 15 mL / kg, and the control group received the same amount of normal saline;

[0028] (2) All rats were sacrificed 24 hours after being exposed to the contrast medium, and kidney tissue was collected, and blood samples were collected from the orbital vein at baseline and 24 hours after administration; the serum creatinine (Scr) concentration in the blood samples was detected. (results such as figure 1 A). figure 1 The results of A show that the Scr concentration of the rats in the experimental group has increased by 56.81% on average, indicating that the Scr concentration of the rats in the experimental group has ...

Embodiment 2

[0030] Example 2. Detection of lncRNAs profile in kidney tissue of CI-AKI rats by next-generation sequencing technology

[0031] (1) Extraction and purification of RNA: TRizol reagent (Invitrogen, Carlsbad, CA) was used to extract total RNA in kidney tissue of CI-AKI rats according to conventional methods. Ribo-Zero rRNA Removal Kit (Epicentre) was used to remove ribose RNA according to the instructions to purify total RNA.

[0032](2) Analysis of RNA purity and quality: measure the concentration and purity of total RNA with a NanoDrop 2000 spectrophotometer (Thermo FisherScientific, Wilmington, DE). When OD260 / OD280 is 1.8-2.2, it is considered that the concentration of total RNA is consistent with RNA Sequencing standard. And check the quality of RNA sample with Agilent 2100 bioanalyzer and RNA 6000 nanometer kit (Agilent, Santa Clara, CA, USA). Samples with an RNA integrity of 8 or higher were considered acceptable, and acceptable RNA was stored at -80°C.

[0033] (3) Us...

Embodiment 3

[0037] Example 3. Reverse transcription quantitative real-time polymerase chain reaction (Reverse transcription quantitative real-time polymerase chain reaction, RT-qPCR) was used to verify the expression of 14 lncRNAs in kidney tissue and whole blood of CI-AKI rats. Proceed as follows:

[0038] (1) Referring to the above RNA extraction method, extract RNA from rat kidney tissue, and use PrimeScript TM RT Reagent Kit (RR037) randomly selects 6mers and oligo-dT primers (Takara, Japan) to reverse transcribe RNA into cDNA;

[0039] (2) Design primers for the above 14 lncRNAs respectively, using GAPDH as an internal reference, the primer sequences of lncRNAs and GAPDH are shown in Table 1.

[0040] (3) in TB Premix Ex Taq TM II (TliRNaseH Plus, RR820) performs quantitative real-time PCR, adding fluorescent groups to the PCR reaction system, and using the accumulation of fluorescent signals to monitor and continuously analyze the entire PCR process in real time;

[0041] (4) D...

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Abstract

The invention mainly provides application of an long-chain non-coding RNA (lncRNA) biomarker to diagnosis of the contrast-induced acute kidney injury (CI-AKI). By illumina sequencing and RT-qPCR, twolncRNAs with expression difference in whole blood of a CI-AKI patient are screened out to serve as the diagnosis biomarkers for the CIAKI. The lncRNA is lnc-HILPDA and lnc-PRND, is remarkably expressed in the whole blood of the CI-AKI patient, is high in prediction efficiency, and has relatively high sensitivity and accuracy. Furthermore, the invention discloses a CI-AKI diagnostic kit which can be prepared from the two lncRNA markers. The CI-AKI diagnostic kit provides a powerful molecular biological tool for CI-AKI diagnosis, and has profound clinical significance and popularization and application prospects.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to the application of two kinds of long-chain non-coding RNA biomarkers in the diagnosis and treatment of contrast agent-induced acute kidney injury, and the two lncRNAs are lnc-HILPDA and lnc-PRND respectively. Background technique [0002] Contrast-induced acute kidney injury (CI-AKI) refers to the decline of renal function within a few days after intravascular administration of iodinated contrast medium, and is a common complication of cardiac catheterization. To date, kidney damage from contrast media is estimated to affect millions of patients, adding to the enormous burden of disease. Therefore, early diagnosis of CI-AKI is very important. At present, researchers widely use serum creatinine as the detection index of acute kidney injury. The KDIGO working group proposed the term "contrast agent-induced acute kidney injury" and suggested a 1.5-fold increase in serum creatinine within 7...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883
CPCC12Q1/6883C12Q2600/158C12Q2600/178
Inventor 刘勇刘博文陈冠仲陈世群陈纪言谭宁
Owner GUANGDONG GENERAL HOSPITAL
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