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New application of S100 calcium binding protein A8 in preparation of airway smooth muscle dilating medicine

A technology of airway smooth muscle and calcium binding protein, which is applied in the field of asthma treatment drugs, achieves good clinical application prospects, safe use, and the effect of relaxing airway smooth muscle

Active Publication Date: 2014-09-24
SHANGHAI UNIV OF T C M +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, some studies have also found the anti-inflammatory activity of S100A8 protein, indicating that S100A8 may play a more complex biological regulatory function under different pathophysiological conditions
Our research found that S100A8 has the function of relaxing airway smooth muscle and reducing airway resistance, but there is no report about this function at home and abroad. This new function may provide a new method for the prevention and treatment of asthma

Method used

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  • New application of S100 calcium binding protein A8 in preparation of airway smooth muscle dilating medicine
  • New application of S100 calcium binding protein A8 in preparation of airway smooth muscle dilating medicine
  • New application of S100 calcium binding protein A8 in preparation of airway smooth muscle dilating medicine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Recombinant expression and purification of S100A8 protein

[0032] 1.1 Amplification of S100A8DNA

[0033] According to the NCBI tissue distribution information, normal rat lung tissue was selected, total RNA was extracted with guanidine isothiocyanate in one step, and cDNA was synthesized by AMV reverse transcriptase, which was used as a template for PCR amplification of S100A8 gene, and PCR amplification was carried out under the action of DNA taq enzyme. The primers for PCR amplification are:

[0034]

[0035] The PCR amplification program is as follows: the reaction system with a total amount of 20ul is pre-denatured at 94°C for 5min, then denatured at 94°C for 30s, annealed at 58°C for 30s, and extended at 72°C for 60s, for a total of 35 cycles, and finally extended at 72°C for 5min; The product sequence is shown in SEQ ID NO:3. It is 100% matched with the rat S100A8 gene in the NCBI database, and the sequence is completely correct.

[0036]1.2 Expr...

Embodiment 2

[0041] Embodiment 2 S100A8 recombinant protein influences the respiratory function of asthmatic rats

[0042] 1. Experimental materials

[0043] 1.1 Experimental objects

[0044] SPF grade male SD rats, weighing 200-300 g, were randomly divided into 6 groups, 8 rats in each group.

[0045] 1.2 Experimental reagents

[0046] 1) Intraperitoneal injection of sensitization solution containing 1mg OVA and 10mgAl(OH) 3 1ml of normal saline was injected intraperitoneally into each rat.

[0047] 2) The challenge solution is physiological saline solution containing 0.5% OVA, and the challenge is injected into the external jugular vein at a dose of 5 mg / kg.

[0048] 3) Other reagents are obtained by preparing analytically pure pharmaceutical saline.

[0049] 4) S100A8 protein, prepared in Example 1.

[0050] 2. Experimental method

[0051] 2.1 Preparation of asthma model

[0052] SD rats were sensitized by intraperitoneal injection of ovalbumin (OVA), and the SD rats were challe...

Embodiment 3

[0066] Example 3 In vitro tissue experiment of the effect of S100A8 recombinant protein on rat tracheal coil tension

[0067] 1. Experimental materials

[0068] 1.1 Experimental objects

[0069] Rat tracheal spiral strip preparation method (rotary cutting method): use pentobarbital sodium to anesthetize SPF grade male SD rats by intraperitoneal injection of 50 mg / kg body weight, dissect out the rat trachea, quickly peel off the surrounding connective tissue, and fix the blade to keep With the blade pointing up, use a metal rod (diameter 2mm) to penetrate the trachea, and press the trachea horizontally on the blade. The spiral is cut into a spiral strip of about 3mm*20mm.

[0070] 1.2 Experimental reagents

[0071] The S100A8 protein (prepared in Example 1) was prepared with pH 7.4 PBS buffer solution to prepare a reagent with a concentration of 1.5ug / ml for use.

[0072] 2. Experimental method

[0073] After the tracheal strips were balanced, different doses of negative c...

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Abstract

The invention relates to the field of a protein medicine, and in particular discloses application of S100 calcium binding protein A8 in preparation of an airway smooth muscle dilating medicine and a pharmaceutical preparation containing the protein. The invention discloses a new function of S100 calcium binding protein A8 for dilating airway smooth muscle; S100A8 can significantly dilate the airway smooth muscle and reduce airway resistance of an asthma model rat so as to treat asthma; S100A8 is applicable to treatment of asthma disease and preparation of an asthma treating medicine; S100A8 is used for treating the asthma disease in a form of injection or spray, and is safe to use, effective and stable.

Description

technical field [0001] The invention relates to the field of protein drugs, and specifically discloses the use of an S100 calcium-binding protein A8 for preparing an asthma treatment drug and the asthma treatment drug containing the recombinant protein. Background technique [0002] S100A8 protein is an important member of the S100 protein family, also known as calgranulin-A (Calgranulin-A), migration inhibitory factor-related protein 8 (Migration inhibitory factor-related protein8), cystic fibrosis antigen (Cystic fibrosis antigen), Leukocyte L1 complex light chain protein (Leukocyte L1 complex light chain), urinary stone protein A (Urinary stone protein band A), etc. The protein is widely expressed in the early stages of cell differentiation in the bone marrow and in circulating granulocytes, monocytes, and activated macrophages, as well as in non-myeloid cells such as endothelial cells and smooth muscle cells. Studies have shown that different expression levels of S100A8...

Claims

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Application Information

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IPC IPC(8): A61K38/17A61P11/08A61P11/06
Inventor 杨永清徐玉东王宇尹磊淼
Owner SHANGHAI UNIV OF T C M
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