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Use of G-protein-coupled receptor kinase 2

A technology of protein-coupled receptors and kinases, applied in medical preparations containing active ingredients, cardiovascular system diseases, respiratory system diseases, etc., can solve problems such as unclear effects and mechanisms

Pending Publication Date: 2018-12-18
陈绍良
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In addition, studies suggest that Berberis plant extracts can improve monocrotaline-induced pulmonary hypertension in rats, but the specific effect and mechanism are still unclear, and further research is needed

Method used

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  • Use of G-protein-coupled receptor kinase 2
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  • Use of G-protein-coupled receptor kinase 2

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Expression of GRK2 in normal donor lungs and pulmonary arteries of PAH patients

[0037] Using healthy donor lungs and PAH patient lung tissues (donated to the First People's Hospital of Wuxi City, Jiangsu Province), the pulmonary arteries were isolated and proteins were extracted for SDS-PAGE-immunoblotting (Western blot), combined with antibodies that specifically recognize GRK2 For detection, β-actin was used as an internal reference. Test results such as figure 1 As shown, GRK2 was significantly upregulated in the pulmonary arteries of PAH patients.

Embodiment 2

[0038] Example 2 Expression of GRK2 in lung tissues of wild-type rats and PAH rats

[0039] (1) Establishment of PAH rat model: SPF grade SD rats (200-250 g) were purchased from the Experimental Animal Center of Nanjing Medical University. The rats were randomly divided into two groups, the control group and the hypoxic group. All were given a normal diet, and the hypoxic group was given a subcutaneous injection of SU5416 (20 mg / kg). After the injection, the animals were placed in a hypoxic box (10% O 2 ). Taken after 4 weeks. The lung tissue was put into corresponding cryopreservation tubes, quickly placed in a liquid nitrogen tank, and stored in a -80°C refrigerator for molecular biology testing.

[0040] (2) The lung tissues of wild-type rats and PAH rats were selected, and proteins were extracted for SDS-PAGE-Western blot test (Western blot), combined with antibodies that specifically recognized GRK2 for detection, and β-actin was used as an internal reference. Test re...

Embodiment 3

[0041] Example 3 Expression of GRK2 in hypoxia-induced pulmonary artery smooth muscle

[0042] (1) Isolation of Pulmonary Artery Smooth Muscle Cells: Clip part of healthy donor Pulmonary Artery under sterile conditions, remove the surrounding tissue, remove the intima and adventitia of the blood vessel, divide the blood vessel into 1.5×1.5mm fragments, and add smooth muscle after 2 hours of attachment Cell culture medium (Sciencell), the medium was changed after 3-4 days, and the tissue pieces were removed after one week, and the cells were used for passage.

[0043] (2) Select passage 3-6 pulmonary artery smooth muscle cells, add serum-free DMEM / F12 starved smooth muscle cells for 12 hours to synchronize the cells, and then give normoxia (20%) and hypoxia (3%) to culture for different time (12h, 24h, 36h, 48h), protein was extracted for SDS-PAGE-Western blot test (Western blot), combined with an antibody that specifically recognizes GRK2 for detection, and β-actin was used as...

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Abstract

The invention belongs to the field of pulmonary hypertension and heart failure treatment, and provides a use of a G-protein-coupled receptor kinase 2, particularly a use of the G-protein-coupled receptor kinase 2 in preparation of products treating the pulmonary hypertension and heart failure. The berberine is adopted as a GRK2 target to inhibit expression and activity of the G-protein-coupled receptor kinase 2, and the G-protein-coupled receptor kinase 2 can be used for treating the pulmonary hypertension and heart failure and can effectively evaluate treatment effects of the berberine.

Description

technical field [0001] The invention belongs to the field of treating pulmonary hypertension and heart failure, and specifically relates to the use of G-protein coupled receptor kinase 2. Background technique [0002] Pulmonary arterial hypertension (PAH) is a chronic pulmonary obstructive disease characterized by intimal hyperplasia and fibrosis of pulmonary vessels, adventitia thickening with moderate inflammation infiltration, thrombus in situ and plexiform lesions . The continuous proliferation and remodeling of small and medium blood vessels in the lung lead to gradual occlusion of the lumen, progressive increase in pulmonary vascular resistance, resulting in right ventricular hypertrophy, eventually leading to right heart failure and even death. The long-term prognosis of PAH is not optimistic. After PAH is diagnosed by right heart catheterization, the 1-, 3-, 5-, and 7-year survival rates are 85%, 68%, 57%, and 49%, respectively. Seeking effective intervention meas...

Claims

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Application Information

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IPC IPC(8): A61K31/4375A61P11/00A61P9/12
CPCA61K31/4375A61P9/12A61P11/00
Inventor 陈绍良顾月
Owner 陈绍良
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