Applications of growth hormone release hormone agonist in preparation of anti-vascular calcification drugs
A growth-stimulating hormone, anti-vascular calcification technology, applied in the fields of pharmacology and chemical biology
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Embodiment 1
[0034] Example 1: Detection of the effect of GHRH-A on smooth muscle cell calcification in vitro
[0035] Implementation process: 6-8 week old C57BL / 6 male mice (purchased from Shanghai Slack Animal Experimental Animal Co., Ltd.) were sacrificed after isoflurane anesthesia to obtain thoracic aorta, digested by collagenase and elastase, plated and cultured, and selected The smooth muscle cells of passage 3-5 were used as experimental objects. Induction in 12-well plate, divided into: (A) control group, (B) osteogenic medium group, (C) osteogenic medium + GHRH-A group, (D) osteogenic medium + GHRH-A - GHRH-Anta. GHRH-A and GHRH-Anta were dissolved in DMSO (Sigma, USA) and stored as 1000x stock solutions. The working concentration of GHRH-A is 10nM, and the working concentration of GHRH-Anta is 100nM. After 14 days of induction, calcification staining, that is, Alizarin Red (Saiye Biotechnology, China) staining was performed and quantified. After the addition of GHRH-A, the s...
Embodiment 2
[0037] Example 2: Detection of the effect of GHRH-A on aortic calcification in vitro
[0038] Implementation process: 8-week-old C57BL / 6 wild-type mice and OPG at the same age - / - Mice (donated by Shan Pengfei’s research group at the Second Affiliated Hospital of Zhejiang University School of Medicine) were grouped as follows: (A) wild-type mice, (B) OPG - / - mouse, (C)OPG - / - mouse+GHRH-A. GHRH-A was dissolved in 0.1% DMSO+10% propylene glycol. The dosage of GHRH-A was 5 μg / d, injected subcutaneously, and intervened continuously for 4 weeks. After 4 weeks, the mice were euthanized, and blood samples and aorta were collected. The aorta was soaked in 10% paraformaldehyde for 24 hours, washed with D-PBS, and scanned by Micro-CT (YUEBO Biotechnology, China). The results showed that there was almost no calcification in the aorta of wild-type mice, while OPG - / - Significant calcified nodules were seen in mice, while in the GHRH-A group, aortic calcified nodules were reduced. ...
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