Function and application of voltage-gated proton channel Hv1 in treating allergic asthma

An allergic asthma, voltage-gated technology, applied in the field of gene function and application, can solve problems such as unclear anti-inflammatory effect

Inactive Publication Date: 2019-12-10
NANKAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although Hv1 has been effectively explored in cancer therapy as we reported, the anti-inflammatory role of Hv1 in asthma remains unclear

Method used

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  • Function and application of voltage-gated proton channel Hv1 in treating allergic asthma
  • Function and application of voltage-gated proton channel Hv1 in treating allergic asthma
  • Function and application of voltage-gated proton channel Hv1 in treating allergic asthma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 The Counting of Different Cells in Alveolar Lavage Fluid of Allergic Mice

[0024] (1) Female mice were randomly divided into the following four groups: normal group (wild type control group=WTC, KO control group=KOC, n=15), model group (wild type model group=WTO, KO model group Group = KOO, n = 15).

[0025] (2) Dissolve 50 μg of OVA in 100 μL of PBS, and then mix it with 100 μL of aluminum for injection to make a sensitization solution. On days 0, 7, and 14, mice in the modeling group were injected intraperitoneally with the sensitizing solution, and mice in the normal group were injected intraperitoneally with a mixture of 100 μL of PBS except OVA and 100 μL of aluminum for injection.

[0026] (3) From day 21, mice in the model group were nebulized with 1% OVA dissolved in PBS for 7 consecutive days, and mice in the normal group were nebulized with PBS, 30 min each time.

[0027] (4) 24 hours after the last nebulization, mice were anesthetized with 50 mg / ...

Embodiment 2

[0029] Example 2 Lung Histopathology of Allergic Mice

[0030] (1) After the mice in Example 1 were euthanized, the lungs were immediately removed under aseptic conditions.

[0031] (2) The right lung was crushed immediately after being frozen in liquid nitrogen, and then stored at -80°C for further use.

[0032] (3) The left lung was fixed with 4% paraformaldehyde for 24 hours. Tissues were dehydrated and embedded in paraffin, cut into 5 μm thick sections for staining.

[0033] (4) The slices were stained with hematoxylin and eosin to detect inflammatory infiltration and vascular congestion, and periodic acid Schiff staining was used to evaluate the proliferation of goblet cells.

[0034] Lung histopathological staining experiments can further evaluate the inflammation of the mouse lung tissue. Compared with the normal group of mice, the modeled mice showed obvious inflammatory cell infiltration, especially around the bronchi and perivascular inflammatory cells. In particu...

Embodiment 3

[0035] Cytokine Detection of Example 3 Allergic Mice

[0036] (1) After diluting the supernatant stored at -80°C in Example 1 to a certain number of times, use the mouse interleukin IL-4 / IL-5 / IL-13 ELISA kit to measure IL-4 and IL-5 in BALF and IL-13 content.

[0037] (2) Extract the total ribonucleotides from the powder stored at -80°C in Example 2 by the phenol-guanidine isothiocyanate method. After reverse transcription into cDNA, the expression of IL-4, IL-5 and IL-13 in lung tissue was detected by real-time fluorescent quantitative PCR.

[0038] The level of Th2 cytokines (IL-4, IL-5 and IL-13) in BALF was measured by enzyme-linked immunosorbent assay, and it was found that compared with the control group mice, the levels of IL-4, IL-5 and IL-13 levels significantly increased. The level of IL-13 in KO OVA control mice was significantly higher than that in WT OVA control mice. The gene expression of Th2 cytokines in lung tissue was similar to the results of ELISA. Rea...

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Abstract

The invention discloses a function of voltage-gated proton channel Hv1 protein in the production of allergic asthma, an established good allergic asthma mouse model, and belongs to the field of medical science. By studying the Hv1 protein, through the comparison with a wild-type C57 (WT) mouse, it is found that the level of allergic airway inflammation in an Hv1 gene knockout mouse (KO) after sensitization and stimulation with ovalbumin (OVA) is significantly higher than that of a control group, that is to say, the Hv1 gene knockout mouse suffers from the allergic asthma. After serum, a bronchoalveolar lavage fluid (BALF) and the pulmonary histopathology are inspected, it is found that Hv1 deficiency significantly promotes the recruitment of OVA-induced inflammatory cells, the level of Th2cytokines of the allergic asthma mouse is increased, and the production of mucus secretion in the allergic asthma mouse is promoted. Through further experiments, it is found that the Hv1 deficiency significantly reduces the expression of NOXs, the results indicate that Hv1 gene knockout increases the prevalence of the allergic asthma by suppressing the yield of ROS. According to the function of the Hv1, the voltage-gated proton channel Hv1 protein can be used for preparing drugs for preventing, alleviating and / or treating the allergic asthma.

Description

technical field [0001] The invention belongs to the field of gene function and application, and in particular relates to the establishment of voltage-gated proton channel Hv1 knockout mice as allergic asthma model mice, the application of Hv1 as a drug target in the screening of drugs for the treatment of allergic asthma, and Hv1 Application of the activator in the preparation and / or screening of drugs for preventing, relieving and / or treating allergic asthma. Background technique [0002] Asthma is one of the major health problems affecting 300 million people worldwide. Morbidity and mortality due to allergic asthma have been increasing over the past few decades. Allergic asthma is a complex pulmonary inflammatory disease characterized by inflammatory cell infiltration into the lung tissue, airway hyperresponsiveness (AHR), goblet cell mucus hypersecretion, and Th2-mediated cytokine (including IL -4, IL-5 and IL-13) overexpression. Th2-type cytokines are critical for the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01K67/027A61K49/00A61K45/00A61P11/06A61P37/08G01N33/535G01N1/30
CPCA01K67/027A01K2227/105A01K2267/03A61K45/00A61K49/0008A61P11/06A61P37/08G01N1/30G01N33/535
Inventor 李树杰杜鸿雁
Owner NANKAI UNIV
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