Use of rno-mir-374 in the preparation of nephrotoxicity biomarkers

A technology of rno-mir-374, 1.rno-mir-374, applied in the field of preparation of nephrotoxic biomarkers, can solve the problems of complex detection methods, inability to detect early detection, and insufficient specificity of nephrotoxic biomarkers, and achieve High feasibility and strong specific effect

Active Publication Date: 2018-02-02
国家(上海)新药安全评价研究中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Therefore, the technical problem to be solved in the present invention is to provide a rno-miR-374 for the preparation of nephrotoxicity biomarkers in view of the problems that the existing nephrotoxicity biomarkers are not specific enough, cannot be detected early, and the detection method is too complicated. Uses and kits and primers in

Method used

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  • Use of rno-mir-374 in the preparation of nephrotoxicity biomarkers
  • Use of rno-mir-374 in the preparation of nephrotoxicity biomarkers
  • Use of rno-mir-374 in the preparation of nephrotoxicity biomarkers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Searching for biomarkers of nephrotoxicity

[0028] 1.1 Establishment of kidney injury model

[0029] 1.1.1 Experimental materials and instruments

[0030] Experimental Reagents and Instruments

[0031] Positive drug: gentamicin sulfate injection (batch number: 20110406, purchased from Hubei Pharmaceutical Co., Ltd., specification: 2mL: 80,000 units; 80mg is 80,000 units);

[0032] Vehicle reference substance: 0.9% sodium chloride injection (normal saline) (batch number 11010463: purchased from Shandong Changfu Crystal Pharmaceutical Co., Ltd.)

[0033] The CRE (R1) (creatinine) assay kit was produced by Wako Pure Chemical Industries, Ltd. (Wako), Japan, and the BUN (urea nitrogen) assay kit was produced by Roche Diagnostics Co., Ltd., Germany.

[0034] HITACHI 7060 automatic biochemical analyzer was purchased from Hitachi Industrial Co., Ltd., Japan.

[0035] experimental animals

[0036] 200 SPF grade SD rats, half male and half male, with body weight b...

Embodiment 2

[0156] Example 2 Detection of the expression level of rno-miR-374 in the blood of rats with renal injury

[0157] In Example 1, three rats administered (80 mg / kg, gentamycin) 7 days later and three rats of the same species growing normally. Extract their total RNA respectively, and extraction method is identical with embodiment 1.

[0158] The extracted total RNA was reverse-transcribed to obtain cDNA (see miScript II RTkit for reverse transcription reagent and system, purchased from Qiagen, Germany). Among them, the reverse transcription system is that each 20 μL reaction solution contains 4 μL of 5╳miScript HiFlex Buffer, 2 μL of 10╳miScript Nucleics Mix, 2 μL of miScript Reverse TranscriptaseMix, and the rest is RNA template and RNase-free water. The program of reverse transcription is: 37°C, 1h; 95°C, 5min; 4°C, keep warm.

[0159] Then detect the expression level of rno-miR-374 by qPCR (see KAPA for qPCR reagents and systems FAST qPCR Kit, purchased from Kapa Biosyste...

Embodiment 3

[0162] Example 3 Detection of the expression level of rno-miR-374 in kidney tissue of rats with renal injury

[0163] Get the kidney tissue of administration rat and normal rat in embodiment 1, extract RNA respectively, reverse transcribe to obtain cDNA, detect the expression level of rno-miR-374 in the kidney tissue by qPCR (extract RNA, reverse transcribe to obtain cDNA , the method detected by qPCR is the same as in Example 2).

[0164] The results showed that the results in Example 3 were similar to those in Example 2, indicating that rno-miR-374 could be used as a renal toxicity biomarker to detect renal injury caused by exogenous compounds.

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Abstract

The invention discloses a use of rno-miR-374 in preparing renal toxicity biomarkers, a kit and primers. rno‑miR‑374 can be used alone as the sole biomarker of nephrotoxicity or in combination with conventional biomarkers of nephrotoxicity and has a role in detecting renal injury induced by exogenous compounds. The kit can detect nephrotoxic biomarkers with strong specificity and high feasibility, so as to detect renal injury caused by exogenous compounds. The primers can specifically and strongly amplify rno-miR-374, and are applied in the kit to detect kidney damage caused by exogenous compounds.

Description

[0001] This application claims the priority of a Chinese patent application with an application date of July 10, 2014, an application number of 201410328083.7, and an invention title of "a nephrotoxicity biomarker and its use". This application cites the full text of the Chinese patent application. technical field [0002] The invention belongs to the field of biotechnology, and specifically relates to a use of rno-miR-374 in the preparation of nephrotoxic biomarkers, a kit and primers. Background technique [0003] The kidney is the most important excretory organ in the body, and it is also one of the main target organs of exogenous poisons in the body. Many compounds (drugs) enter the body and can cause nephrotoxicity. According to investigations, 32.4% of acute renal failure is caused by drug nephrotoxicity. Therefore, there is an urgent need to establish, discover and confirm new biomarkers of nephrotoxicity, which can detect renal injury early, and can reflect the impr...

Claims

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Application Information

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Patent Type & AuthorityPatents(China)
IPC IPC(8): C12N15/113C12Q1/68C12N15/11
Inventor邱云良马璟洪敏富欣汤纳平李华黄欢夏
Owner国家(上海)新药安全评价研究中心