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Urine protein marker for chronic pancreatitis and application thereof

A chronic pancreatitis and protein technology, applied in the field of protein markers, can solve the problems of low sensitivity of pancreatic function test and inability to provide valuable diagnostic clues

Inactive Publication Date: 2018-03-20
BEIJING NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Imaging information cannot provide valuable diagnostic clues at an earlier stage of the disease, the sensitivity of pancreatic function tests is low, and pancreatic biopsy is a definitive standard for the diagnosis of CP, but biopsy itself has certain defects

Method used

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  • Urine protein marker for chronic pancreatitis and application thereof
  • Urine protein marker for chronic pancreatitis and application thereof
  • Urine protein marker for chronic pancreatitis and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Embodiment 1: Establishment of chronic pancreatitis rat model

[0050] Diethyldithiocarbamate (diethyldithiocarbamate, DDC) induced chronic pancreatitis rat model fibrosis can simulate the process of chronic pancreatitis disease (see Naoki Matsumura et al., Study onFree Radicals and Pancreatic Fibrosis-Pancreatic Fibrosis Induced by Repeated Injections of Superoxide Dismutase Inhibitor Panereas, 2001, 22; 53-57). Observe the overall changes of pancreatic fibrosis from normal, early fibrosis, fibrosis progression, and its pathological process and pathological features are similar to human chronic pancreatitis. Intraperitoneal injection of DDC induced pancreatic injury in rats in a gradual process with good reproducibility. Using this animal model to simulate the pathogenesis of chronic pancreatitis has important guiding significance for clinical early diagnosis and monitoring of disease progression.

[0051] In the present application, the animal model of rat chronic p...

Embodiment 2

[0068] 1. Urinary protein extraction and preservation:

[0069] The urine was centrifuged at 2000g at 4°C for 30 minutes, the supernatant was taken, placed in a new EP tube, and centrifuged at 12000g at 4°C for 30 minutes; the supernatant was taken and stored at -80°C.

[0070] Urine protein was precipitated with ethanol, and the protein concentration was measured by Bradford method, followed by enzyme digestion on the membrane. See WisniewskiJR, Zougman A, Nagaraj N, Mann M. Universal sample preparation method for proteome analysis. Nature methods 2009; 6:359-62. The BCA method was used to measure the concentration of peptides.

[0071] 2. LC-MS / MS tandem mass spectrometry analysis:

[0072] Peptide samples were diluted to 0.5 μg / μl with 0.1% formic acid. Peptide samples were separated by Thermo liquid phase system EASY-nLC 1200 loading system. The elution time is 120 minutes, and the column flow rate is 0.3 μl / min. The elution gradient was 5% to 40% mobile phase B (mobil...

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Abstract

The present application relates to urinary protein markers for chronic pancreatitis and uses thereof. Specifically, this application relates to the use of urine protein markers obtained by using chronic pancreatitis disease models and mass spectrometry analysis in the early diagnosis of human chronic pancreatitis disease and the use of disease course monitoring. The urine protein markers are selected from Igγ‑1 chain C domain, nucleonectin‑2, beta‑2‑microglobulin, interleukin‑4 receptor alpha subunit, phosphoglycerate mutase 1, dipeptidyl peptidase 2, secreted pyrophosphoprotein 24, glycans Core protein, endothelial cell selective adhesion molecule, nestin-2, serum amyloid substance p, nerve cell adhesion molecule 1, etc. In this application, the obtained differential protein in urine provides a non-invasive and convenient option for early diagnosis and subsequent treatment of chronic pancreatitis.

Description

technical field [0001] This application relates to clinical medicine, in particular to protein markers related to human chronic pancreatitis. Specifically, the present application relates to the use of protein markers related to the diagnosis and condition monitoring of human chronic pancreatitis obtained by using the rat chronic pancreatitis disease model and mass spectrometry analysis. Background technique [0002] Biomarkers are monitorable changes associated with physiological or pathological processes. The study of disease markers has always been a research focus and difficulty in recent years. Appropriate markers mean that they can indicate the occurrence and development of the disease earlier and more sensitively, and provide accurate information for the diagnosis, treatment and prognosis of the disease, which is more beneficial to the health of the patient. [0003] The essence of biomarkers is change. Urine, as the excrement excreted by the body through the urinar...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/6893G01N2800/067
Inventor 高友鹤张琳珮李育秋
Owner BEIJING NORMAL UNIVERSITY
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