Application of substances that inhibit phosphorylation of 357th tyrosine residue of yap protein in preventing atherosclerosis
A technology for atherosclerosis and products, which is applied in the field of biomedicine and can solve problems such as the unclear interpretation of signal transduction mechanisms
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Embodiment 1
[0030] Example 1, Src / c-Abl dual inhibitor inhibits activation of vascular endothelial cells
[0031] Both Nilotinib and Bosutinib are Src / c-Abl dual inhibitors, and Bosutinib is the third-generation Src / c-Abl dual inhibitor.
[0032] 1. Src / c-Abl dual inhibitor significantly affects YAP tyrosine phosphorylation
[0033] 1. Take the cell plate, add fibronectin, and incubate overnight at 4°C.
[0034] 2. After completing step 1, take the cell plate, discard fibronectin, and inoculate 3×10 6 HUVECs, and then add Nilotinib-containing DMSO solution or Bosutinib-containing DMSO solution to obtain a treatment system. In the treatment system, the concentration of Nilotinib or Bosutinib was 10 μM.
[0035] 3. After completing step 1, take the cell plate, discard fibronectin, and inoculate 3×10 6 HUVECs, then add DMSO (equal volume to the DMSO solution containing Nilotinib in step 2) to obtain a treatment system (as a control).
[0036] 4. After completing steps 2 and 3, take the tr...
Embodiment 2
[0050] Example 2, p-YAP Y357 and p-c-Abl Y412 Important role in vascular endothelial cell activation and plaque formation
[0051] 1. Detection of p-c-Abl in mouse vascular arch Y412 and p-YAP Y357 expression level of
[0052] 1. Take 6-8 weeks old ApoE respectively - / - The vascular arches (bifurcation and lnner) of mice are prone to atherosclerosis, and the blood flow is turbulent, referred to as AA) and the straight part of the blood vessel (the In the protected area, the form of blood flow is laminar flow, referred to as TA).
[0053] 2. After completing step 1, perform immunofluorescence staining to determine the phosphorylation levels of YAP and c-Abl.
[0054] For the experimental results of the phosphorylation level of YAP tyrosine 357, see figure 2 Middle A (green fluorescently labeled p-YAP Y357 , red fluorescent marker CD31 (endothelial cell marker), blue marker nuclei (DAPI), the scale bar is 80 μm, n=6, merge is superposition effect). The experimental res...
Embodiment 3
[0062] Example 3, Bosutinib slows diet and partial ligation-induced atherosclerosis
[0063] Western Diet is a product of Research Diets, Inc., catalog number D12109C.
[0064] 1. Bosutinib slows down diet-induced atherosclerosis
[0065]To further verify the role of tyrosine kinase-mediated YAP phosphorylation in promoting atherosclerotic phenotype in vascular endothelial cells, the inventors of the present invention conducted the following experiments.
[0066] 1. Take 11 6-week-old ApoE mice - / - The mice were randomly divided into an experimental group and a control group, with 5 mice in the experimental group and 6 mice in the control group. Do the following:
[0067] Experimental group: DMSO solution containing Bosutinib was injected intraperitoneally one week before the experiment, with a dose of 30 mg Bosutinib / kg per injection, once every two days for 5 weeks; from the second week, Western diet was added for 4 weeks.
[0068] Control group: The same volume of DMSO ...
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