Application of heat shock protein 20 in oxidative stress injury diseases

A technology of heat shock protein and oxidative stress, which is applied in the field of application of heat shock protein 20 in oxidative stress injury diseases, can solve problems that have not been reported, and achieve prevention of oxidative stress injury, high clinical application prospects, delay Effects of disease progression

Inactive Publication Date: 2018-08-10
SHANGHAI FIRST PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

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  • Application of heat shock protein 20 in oxidative stress injury diseases
  • Application of heat shock protein 20 in oxidative stress injury diseases
  • Application of heat shock protein 20 in oxidative stress injury diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Embodiment 1: animal experiments

[0049] 1. Materials and methods

[0050] 1. Animal grouping and model preparation

[0051]BALB / c mice: special pathogen-free grade, female, 8-10 weeks old, weighing about 20 g, purchased from Shanghai Shrek Experimental Animal Center. All mice were kept in the independent ventilation system (IVC) of the Animal Experiment Center of the First People's Hospital Affiliated to Shanghai Jiaotong University, with an ambient temperature of 22°C-24°C, a humidity of 40-60%, 12-hour light control, and free access to food and water. All operations and experimental procedures were approved by the Experimental Animal Ethics Committee of the First People's Hospital Affiliated to Shanghai Jiaotong University.

[0052] 20 animals were randomly divided into 4 groups, 5 animals in each group:

[0053] Normal + air exposure group: exposed to filtered air on the 1st, 3rd, 5th, and 7th days, each time for 1 hour, and exposed to filtered air for 3 hours o...

Embodiment 2

[0080] Example 2: Cell experiment

[0081] 1. Materials and methods

[0082] 1. Cells

[0083]Human airway epithelial cell line BEAS-2B cells (purchased from ATCC) were cultured in vitro. RPMI1640 culture medium containing 10% fetal bovine serum was used.

[0084] 2. Determine H 2 o 2 Stimulus concentration and observation time

[0085] Cells were incubated in a 24-well plate, given gradually increasing H 2 o 2 Concentrations (0.1, 1.0, 10, 100, μM) were stimulated for 1 hour, and the cell viability was detected by MTT at 0, 12 and 24 hours respectively, and the expression of HSP20 protein and gene level were detected by Western blot and RT-qPCR.

[0086] 3. Functional experiment of HSP20 and its phosphorylated protein

[0087] The cells were transfected with pEX-3-HSP20 (wild type overexpression vector), pEX-3-HSP20(S16A) (non-phosphorylated overexpression vector) and pEX-3 (empty vector). Refer to the specific process 2000 (purchased from ThermoFisher Scientific C...

Embodiment 3

[0098] Embodiment 3: Molecular biology detection

[0099] 1. Western blot detection

[0100] After weighing the lung tissue, add the lysate according to the volume of 1:10, then carry out shear dispersion and emulsification on ice, and centrifuge the lung tissue homogenate (or cell lysate) at 13,000 rpm for 15 minutes at 4°C; take 100 μl of the supernatant and add it to the After the sample buffer was boiled at 95°C for 15 minutes, centrifuged slightly and placed at -80°C for testing, another 5 μl of the homogenate supernatant was used to detect the protein concentration by the BCA method. Protein electrophoresis was performed using 10% SDS-PAGE. Load 75 μg of each sample, and determine the loading volume according to the protein concentration. 80V voltage for 30 minutes, 120V voltage for 90 minutes, semi-dry electrotransfer membrane, 20V for 7 minutes. After the transfer was completed, the transfer efficiency was judged by Ponceau red pre-staining. After rinsing with 1×TBS...

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Abstract

The invention relates to novel application of heat shock protein 20(HSP20), in particular to application of the HSP20 in preparing a drug for preventing and treating oxidative stress injury diseases.The invention further provides a building method of a mouse with high HSP20 expression in an AEC. The HSP20 has the advantages that it is found for the first time that HSP20 can be expressed when theAEC is subjected to oxidative stress and has an oxidation resistance effect, the expression of the HSP20 in the AEC is improved, the oxidation resistance of the AEC itself is improved, then the occurrence of the oxidative stress injury is prevented, and the HSP20 can be used for resisting ill effects of oxidative stress relevant to chronic air flue diseases, and has a very good clinical application prospect. By means of the novel application of the heat shock protein 20(HSP20), it is found for the first time that the HSP20 can be subjected to oxidative stress inducible expression, and has an oxidation resistance protection effect. It is found for the first time that the mechanism of the oxidation resistance protection effect of the HSP20 is relevant to regulation of an Nrf2-NQO-1 signal channel by the HSP20.

Description

technical field [0001] The invention relates to the technical field of medicines, in particular to the application of heat shock protein 20 in oxidative stress injury diseases. Background technique [0002] Oxidative stress is a very important driving factor in the occurrence and development of chronic airway diseases, and the oxidative stress injury of airway epithelial cells (AEC) is the central link. However, some current antioxidant treatments (including food and drugs) try to reduce oxidative stress damage by neutralizing oxidative stress metabolites such as reactive oxygen species (ROS) by directly administering substances with antioxidant activity. Clinical trials have shown that none of these methods can prevent oxidative stress damage in AEC well. Therefore, it is necessary to discover the mechanism that can enhance the antioxidant capacity of airway epithelial cells, and then reduce the formation of oxidative stress metabolites from the inside of the cells and fro...

Claims

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Application Information

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IPC IPC(8): A61K38/17A61K45/00A61P39/06A61P11/00A61P11/06A61K33/00
Inventor 包爱华张旻贲素琴杨红乔丽华包婺平周新
Owner SHANGHAI FIRST PEOPLES HOSPITAL
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