Application of GPR31 inhibitor in preparation of medicine for preventing and treating vascular calcification

A vascular calcification and inhibitor technology, applied in the field of medicine, to achieve the effect of broad application prospects

Pending Publication Date: 2022-08-05
HEFEI UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

GPR31 is more discussed, but less studied in vascular calcification

Method used

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  • Application of GPR31 inhibitor in preparation of medicine for preventing and treating vascular calcification
  • Application of GPR31 inhibitor in preparation of medicine for preventing and treating vascular calcification
  • Application of GPR31 inhibitor in preparation of medicine for preventing and treating vascular calcification

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1: Application of GPR31 inhibitors in the preparation of drugs for preventing and treating vascular calcification

[0026] In the experimental study of the present invention, RNA-seq sequencing of whole blood samples found that GPR31 was most significantly up-regulated in 4 patients with coronary artery calcification compared with 4 control patients. In human carotid atherosclerotic plaques, GPR31 expression was significantly elevated in calcified plaques. The above results indicate that GPR31 may be involved in the occurrence and development of coronary calcification.

[0027] GPR31 expression can be detected in circulating leukocytes derived from patients with coronary calcification, and the expression level is close to that in monocytes (THP1), macrophages (Macrophage), endothelial cells (HUVEC) and smooth muscle cells (HASMC) . After calcification was induced in HASMC cells with high phosphate medium, the expression of GPR31 was up-regulated, indicating th...

Embodiment 2

[0065] Example 2: GPR31 protein expression in different cells.

[0066] The proteins of circulating leukocytes, endothelial cells, monocytes, macrophages, smooth muscle cells and calcified smooth muscle cells in patients with coronary calcification were extracted, and the expression of GPR31 in different types of cells was analyzed by Western blot.

[0067] Western blot:

[0068] 1) Add pre-cooled protein lysate (pre-add protease inhibitor) after cell treatment. The cells were pipetted evenly, transferred to a 1.5mL centrifuge tube, placed on ice for 5min reaction, fully shaken and then reacted on ice for 5min, centrifuged at 4°C, 12000rpm for 10min, collected the supernatant, and stored at -20°C for a long time in the refrigerator.

[0069] 2) Determination of protein concentration using BCA method.

[0070] 3) SDS-polyacrylamide gel electrophoresis (SDS-PAGE): SDS-PAGE operation steps mainly include: preparation of polyacrylamide gel, preparation of sample loading system,...

Embodiment 3

[0077] Example 3: Expression of GPR31 in the aortic root and the initial segment of the coronary artery in the control and coronary calcification mouse models.

[0078] LDLR - / - Mice were fed a high-fat diet for 16 weeks to establish a coronary calcification model. The aortic roots and hearts of mice were collected, and frozen sections were used to locate the aortic roots and coronary arteries. Alizarin red staining was performed on the aortic roots and coronary sections to detect Calcium deposition. The expression of GPR31 was detected by immunofluorescence. The steps of tissue alizarin red staining and tissue immunofluorescence were described in Example 1.

[0079] like image 3 shown, image 3 Medium, LDLR - / - Mice were fed a high-fat diet for 16 weeks, aortic roots and hearts were collected, and frozen sections were used to locate the aortic roots and coronary arteries. A: Alizarin red staining was performed on aortic root and coronary sections to detect calcium depo...

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Abstract

The invention discloses an application of a GPR31 inhibitor in preparation of a medicine for preventing and treating vascular calcification. The vascular calcification is a common pathological expression commonly existing in atherosclerosis, hypertension, diabetes, vasculopathy, chronic kidney disease, senescence and the like. According to the invention, the inhibition effect on the GPR31 gene is realized by utilizing siGPR31, and smooth muscle cell osteogenic differentiation induced by a high phosphate culture medium can be remarkably treated, so that the GPR31 inhibitor has a relatively wide application prospect in treatment of vascular calcification.

Description

technical field [0001] The invention relates to the application of a GPR31 inhibitor in the preparation of a drug for preventing and treating vascular calcification, and belongs to the technical field of medicine. Background technique [0002] Vascular calcification is a common pathological manifestation of atherosclerosis, hypertension, diabetic vascular disease, vascular injury, chronic kidney disease and aging. [0003] Coronary calcification is a complex, active process that partly resembles bone mineralization. The causes and mechanisms of coronary calcification are complex, including hyperphosphatemia, vascular smooth muscle cell (VSMC) osteogenic transformation, matrix vesicle release, apoptosis, inflammation, oxidative stress, endoplasmic reticulum stress, etc. . At present, the molecular mechanism of coronary calcification is not fully understood, especially the endogenous promoting factors of coronary calcification are still unclear. Thus, there is also a lack o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/00A61K31/713A61P9/14A61P9/10
CPCA61K45/00A61K31/713A61P9/14A61P9/10
Inventor 陈元利朱猛猛段亚君韩际宏马礼坤胡昊潘健源
Owner HEFEI UNIV OF TECH
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