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Application of CD146 as treatment target point to preparation of medicines for treating asthma airway remodeling

A therapeutic target and airway remodeling technology, applied in the field of medical testing, can solve the problems such as CD146 that have not yet been seen

Active Publication Date: 2020-06-05
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

However, there is no report on the relationship between CD146 and asthma

Method used

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  • Application of CD146 as treatment target point to preparation of medicines for treating asthma airway remodeling
  • Application of CD146 as treatment target point to preparation of medicines for treating asthma airway remodeling
  • Application of CD146 as treatment target point to preparation of medicines for treating asthma airway remodeling

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1 Detection of plasma CD146 expression levels in asthmatic patients and normal persons

[0058] Main reagents: human CD146 (Melanoma Cell Adhesion Molecule) ELISA box ElabscienceE-EL-H2403c

[0059] Main instruments: microplate reader, centrifuge

[0060] Main methods: The peripheral blood samples of 28 asthmatic patients were obtained from Jiangsu Provincial People's Hospital and Taizhou People's Hospital, and the samples of 30 healthy people were obtained from Jiangsu Provincial Blood Center. After the peripheral blood was collected, it was placed into an anticoagulant tube, and the upper layer plasma was collected by centrifugation at 1000 rpm, and stored at -80°C. The expression of CD146 in human plasma was detected by enzyme-linked immunosorbent assay.

[0061] CD146 was detected by double antibody sandwich ELISA. The anti-human CD146 antibody is used to coat the microtiter plate, and the human CD146 in the sample or standard will bind to the coated anti...

Embodiment 2

[0062] Example 2 HDM induces increased expression of CD146 in bronchoalveolar epithelial cells through IL-33 / ST2

[0063] 2.1 Real-time quantitative pCR detection of CD146 expression of MLE-12 under HDM stimulation

[0064] Main method:

[0065] HDM was dissolved in PBS at a concentration of 10mg / ml and stored at -80°C. 0, 10, 100 μg / ml HDM stimulated MLE-12 for 0, 6, 12, and 24 hours, then used trizol or small RNA to extract the total RNA of MLE-12 cells stimulated by HDM, took an equal amount of RNA and obtained cDNA after reverse transcription, and used After diluting with the corresponding volume of RNase free water, the SYBR Green method was used to detect the expression of CD146 by fluorescent quantitative PCR, and each sample had 3 replicate wells. The expression levels of different transcription factors and cytokines were calculated using the ΔΔCT value of β-actin as the internal reference standard. The primers are as follows: CD146 forward, 5'-GGACCTTGAGTTTGAGTGG-3...

Embodiment 3

[0133] Example 3 HDM induces increased expression of CD146 in bronchoalveolar epithelial cells through the p65 signaling pathway

[0134] 3.1 Western blot detection of expression changes of MyD88, p65, p38, JNK, p44 / 42 signaling pathways after stimulation of MLE-12 cells with HDM

[0135] Main method: The method of HDM stimulating MLE-12 is the same as above. Cells were lysed with Lysis Buffer and centrifuged to extract protein, and Western blot was performed after quantification.

[0136] Main reagents: Lysis Buffer, loading buffer, anti-beta-actin antibody (Cell Signaling Technology, #4970), anti-MyD88 antibody (Cell Signaling Technology, #4283), anti-p65 antibody (Cell Signaling Technology, #8242) , anti-p65 (phospho-Ser536) antibody (Cell Signaling Technology, #3033), anti-p38 (Cell Signaling Technology, #8690), anti-P38 (phospho-Thr180 / Tyr182) antibody (Cell Signaling Technology, #4511), anti -JNK (Cell Signaling Technology, #9252), anti-JNK (phospho-Thr183 / Tyr185) anti...

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Abstract

The invention discloses an application of CD146 as a treatment target point to preparation of medicines for treating asthma airway remodeling, an application of the CD146 as the treatment target pointto screening and treating asthma, an application of substances for restraining CD146 expression or antagonism CD146 to preparation of medicines for treating bronchial asthma, and preferably, an application of preparation of medicines for asthma airway remodeling. The invention discloses an application of the CD146 for detecting plasma of patients suffering from bronchitis. For the first time, theinventor finds expression differences of the CD146 in serum of patients suffering from asthma and serum of normal people. The indexes can be used as a serum molecule marker for the bronchial asthma.The research indicates that HDM can worsen the airway remodeling through the CD146. Therefore, the CD146 can be used as the treatment target point to be used for screening and treating bronchial asthma. In accordance with an antibody of the CD146, the CD146 is also a treatment strategy for the bronchial asthma.

Description

technical field [0001] The invention belongs to the field of medical testing and relates to the application of CD146 as a therapeutic target in the preparation of medicines for treating asthmatic airway remodeling. Background technique [0002] The human adhesion molecule CD146 was first discovered on melanoma cells. It is a single transmembrane glycoprotein composed of 646 amino acids, including a V-VC2-C2-C2 immunoglobulin-like domain, and the complete transmembrane form includes the extracellular region. , transmembrane and cytoplasmic regions. It has been found that CD146 exists in two forms: membrane type and soluble form. Membrane type CD146 molecules are widely distributed in vascular endothelial cells and smooth muscle cells, as well as partially activated lymphocytes, etc., and participate in the development of blood vessels, cell adhesion and migration, immune response and Mesenchymal stem cell differentiation, etc. In addition, CD146 has been reported to be high...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/00A61K39/395A61P11/06C12Q1/6883G01N33/68
CPCA61K39/3955A61K45/00A61P11/06C12Q1/6883C12Q2600/158G01N33/6893G01N2800/122
Inventor 吉宁飞孙知晓马其云陈中琦王正霞黄茂张明顺
Owner JIANGSU PROVINCE HOSPITAL THE FIRST AFFILIATED HOSPITAL WITH NANJING MEDICAL UNIV
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