Application of vascular endothelial cell SIRT6 gene, and drug
A vascular endothelium and drug technology, which is applied to the function and application field of SIRT6 gene in endothelial dysfunction-related diseases, can solve the problem that there is no relevant report on the protective effect, and achieve the effect of protecting endothelial function.
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0029] Example 1 Construction and identification of SIRT6 endothelial-specific knockout mice
[0030] In order to further study the function of SIRT6 deletion in endothelial cells on vascular endothelium, the present invention constructed SIRT6 endothelial-specific knockout mice. First, the female SIRT6 flox / flox Mice were crossed with male C57BL / 6J Tie2-Cre mice to obtain Tie2-Cre / SIRT6 flox / + mice; then male Tie2-Cre / SIRT6 flox / + SIRT6 in mice and females flox / flox Mice were crossed to obtain homozygous SIRT6 endothelial-specific knockout mice, namely ecSIRT6 - / - mice. The present invention selects SIRT6 flox / flox Mice were used as wild-type control mice.
[0031] Identification of SIRT6 endothelial-specific knockout mice: First, the DNA of the mice to be identified was extracted using a kit from Biotool, USA, and identified by agarose gel electrophoresis after PCR amplification. Cut about 3mm of the tail of the mouse to be identified and put it into a 1.5mL EP tube. ...
Embodiment 2
[0035] Example 2 Observation on histomorphology of SIRT6 endothelial-specific knockout mice and SIRT6 endothelial-specific knockout mice with hyperlipidemia
[0036] 1. Material collection: After the experiment, the mice in each group were killed, and the thoracic aorta, carotid artery, heart, liver and other tissues were gently removed with ophthalmic scissors. Wash away the adherent blood with normal saline, and blot dry gently. First fix in 4% paraformaldehyde for 48h, then dehydrate with 15% sucrose solution for 24h, and finally dehydrate with 30% sucrose solution for 24h. After dehydration, tissue embedding was carried out with OTC. The embedded tissue was cut into 7 μm uniform slices with a cryostat, stored in a -80°C refrigerator and subjected to HE staining in the next step.
[0037] 2. Staining: Take out the slices from -80°C, rewarm for 30 minutes; fix with 4% paraformaldehyde for 15 minutes; rinse with PBS for 1 minute, rinse twice; stain with hematoxylin for 10 m...
Embodiment 3
[0038] Example 3 Detection of serum NO content and NOS enzyme activity in SIRT6 endothelial-specific knockout mice
[0039] About 1.5 mL of blood was collected from the mouse orbit, centrifuged at 3500 rpm for 10 min, and the supernatant was collected and stored at -20°C. Using the Nitric Oxide Assay Kit, Nitrate Reductase was used to specifically convert NO 3- Revert to NO 2- , and then reflect the level of NO concentration through its color depth. The NOS enzyme activity kit was used to detect the enzyme activities of tNOS, iNOS and cNOS, respectively. The results are shown in Table 2 and image 3 Shown:
[0040] Table 2
[0041]
[0042] From image 3 It can be seen from the figure that the deletion of SIRT6 in endothelial cells will reduce the content of NO in mouse serum and the enzyme activity of total NOS and cNOS, and the content of NO in the serum of homozygous SIRT6 endothelial-specific knockout mice, the total The activity of NOS and cNOS enzymes decreased...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More