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Application of PPM1A in treatment and diagnosis of asthma

A diagnostic reagent, technology for bronchial asthma, applied in the field of biomedicine

Active Publication Date: 2019-05-03
JIANGSU PROVINCE HOSPITAL THE FIRST AFFILIATED HOSPITAL WITH NANJING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the past, there were many studies on PPM1A in the field of tumors, and it was considered as a tumor suppressor factor. So far, no one has found the relationship between PPM1A and asthma

Method used

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  • Application of PPM1A in treatment and diagnosis of asthma
  • Application of PPM1A in treatment and diagnosis of asthma
  • Application of PPM1A in treatment and diagnosis of asthma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Example 1 Detection of PPM1A protein level in peripheral blood of asthmatic patients

[0057] main reagent

[0058] Human PPM1A Elisa Detection Kit (Wuhan Bio-swamp)

[0059] main instrument

[0060] Microplate reader, centrifuge

[0061] main method

[0062] Collection and freezing of peripheral blood samples

[0063]The peripheral blood samples of 20 asthmatic patients came from Jiangsu Provincial People's Hospital, and the samples of 20 healthy people came from college students of Nanjing Medical University. Collect peripheral blood and anticoagulant tubes from normal people and asthma patients, centrifuge at 1000rpm / min for 5 minutes, collect the upper layer of plasma, and freeze it at -80°C

[0064] Detection of PPM1A expression in human plasma by enzyme-linked immunosorbent assay

[0065] This kit uses the double-antibody sandwich method to determine the level of human Mg2+ / Mn2+-dependent protein phosphatase 1A (PPM1A) in the specimen. Coat the microwell pl...

Embodiment 2

[0078] Example 2 Increase the sample size and re-detect the expression of PPM1A

[0079] main reagent

[0080] Human PPM1A Elisa Detection Kit (Wuhan Bio-swamp)

[0081] main instrument

[0082] Microplate reader, centrifuge

[0083] main method

[0084] Collection and freezing of peripheral blood samples

[0085] The peripheral blood samples of 50 asthmatic patients came from Jiangsu Provincial People's Hospital, and the samples of 50 healthy people came from college students of Nanjing Medical University. Collect peripheral blood and anticoagulant tubes from normal people and asthma patients, centrifuge at 1000rpm / min for 5 minutes, collect the upper layer of plasma, and freeze it at -80°C

[0086] Detection of PPM1A expression in human plasma by enzyme-linked immunosorbent assay

[0087] Same as Example 1

[0088] result

[0089] Plasma PPM1A levels in asthmatic patients were higher than normal people, with statistical significance ( figure 1 B).

Embodiment 3

[0090] Example 3 Regulation of miR-1165-3p on Th2 cell differentiation (cellular level)

[0091] Main reagents:

[0092] Cytokines (peprotech), functional antibodies (ebioscience), serum (Gibco), primers (GenScript), sybr green (takara), miRNA reverse transcription reagents (takara), 1640 medium (Gibco), Elisa kit (Biolegend )Western antibody (Abcam, CST), flow cytometry antibody and reagent (ebioscience), Trizol and small RNA extraction reagent (Thermo), magnetic bead sorting kit

[0093] main instrument

[0094] Cell incubator, magnetic frame, flow detection machine, electrophoresis transfer membrane instrument, exposure instrument, PCR instrument and qPCR instrument, microplate reader

[0095] main method

[0096] Th1 / Th2 cells differentiated in vitro

[0097] 1. Coating: Add 100ul 5ug / ml anti-CD3e to each well, overnight at 4°C.

[0098] 2. Isolation of mouse Naive CD4+ T cells. (Day 0)

[0099] ① 6-8 week old female C57 / B6 mice were killed by cervical dislocation, ...

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Abstract

The invention discloses application of PPM1A in the treatment and diagnosis of asthma. To be specific, the invention relates to application of the PPM1A as a detection target in preparation of a diagnostic reagent for bronchial asthma, application of a regent detecting the PPM1A in preparation of a diagnostic reagent for bronchial asthma, application of the PPM1A as a therapeutic target in preparation or screening of a drug for treating bronchial asthma, and application of a substance suppressing PPM1A expression in preparation of a drug for treating bronchial asthma. The invention discloses the application of PPM1A in detecting the plasma of the bronchial patient; expressions of the PPM1A in an asthma patient and a normal person are discovered to be different for the first time; and the indicator can be used as the molecular marker of the bronchial asthma. Besides, the PPM1A acting as a miR-1165-3p target gene is discovered to be able to promote Th2 differentiation through the STAT1,STAT5, and Akt accesses to cause the asthma, so that the PPM1A can used as a therapeutic target for asthma.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to the application of PPM1A in the treatment and diagnosis of asthma. Background technique [0002] Asthma is a heterogeneous disease characterized by chronic airway inflammation, airway remodeling, and airway hyperresponsiveness. There are about 300 million asthma patients in the world, and the number of asthma patients in my country exceeds 3 million. In recent years, with the progress of industrialization, the prevalence of asthma in my country is still increasing year by year, which has brought heavy family and social burdens. Due to the heterogeneity of asthma, asthma has multiple phenotypes and endotypes. The Global Initiative for Asthma (GINA) 2018 divides asthma into the following phenotypes: allergic asthma, nonallergic asthma, delayed-onset asthma, asthma with fixed airflow limitation, and asthma with obesity. Asthma populations with different phenotypes respond differently to d...

Claims

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Application Information

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IPC IPC(8): G01N33/573A61K45/00A61P11/06
Inventor 黄茂王正霞陈中琦孙知晓马其云张明顺吉宁飞
Owner JIANGSU PROVINCE HOSPITAL THE FIRST AFFILIATED HOSPITAL WITH NANJING MEDICAL UNIV
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