Application of interleukin 37 in regulation of angiogenesis disease
An angiogenesis and interleukin technology, applied in the field of biomedicine, can solve problems such as unsatisfactory effects
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Embodiment 1
[0099] Example 1 IL-37 Promotes the Proliferation and Migration of Endothelial Cells
[0100] 1.1 Proliferation experiment
[0101] The cultured HUVEC were placed in a 96-well plate. After the cells adhered to the wall, different doses of IL37 and VEGF were added. After 12 hours of continuous culture, 10 μL of CCK-8 was added, mixed and cultured for 2 hours to measure the absorbance at 450 nm.
[0102] The results showed that IL-37 had obvious proliferative effect on HUVEC, and it could promote proliferation at a low concentration (0.001ng / ml), and the effect was most obvious at a concentration of 1ng / ml. The pro-proliferation effect of IL-37 was better than that of VEGF( figure 1 a).
[0103] 1.2 Lumen formation
[0104] Spread Matrigel evenly on the bottom of a 96-well plate, and inoculate 80,000 HUVECs per well after solidification. After the cells adhered to the wall, different doses of IL37 and VEGF were added, and the lumen formation of each experimental group was ob...
Embodiment 2
[0113] Example 2 The role of IL-37 in promoting angiogenesis
[0114] 2.1 Arterial ring experiment
[0115] The wild-type mice of about 8 weeks old were taken, the thoracic aorta was taken out, the blood was washed clean, and the connective tissue around the blood vessel was peeled off. Cut blood vessels into arterial rings with a length of about 1 mm, add 50 μl Matrigel to a 96-well plate, and coat the arterial rings in Matrigel. After the Matrigel is solidified, add 150 μl of complete medium containing IL-37, IL-37+IgG, IL-37+IL37antibody, at 37°C, 5% CO 2 cultured in an incubator. After 1 week, observe the sprouting situation of the mouse aorta under a microscope, take pictures, and count the sprouting points of the mouse aorta.
[0116] The results showed that IL-37 can promote mouse aortic sprouting, and the promoting effect can be specifically inhibited by IL-37 neutralizing antibody ( figure 2 a), illustrating that IL-37 promotes endothelial cell proliferation spec...
Embodiment 3
[0123] Example 3 The effect of IL-37 on neovascularization under pathological conditions
[0124] In this example, the oxygen-induced retinal neovascularization model was used to further study the effect of IL-37 on neovascularization under pathological conditions.
[0125] Seven days after birth, the mice were placed in an animal experiment cabin connected with oxygen supply along with two nursing mothers, so that the oxygen concentration was stably controlled at about 75%. 12 days after the birth of the mice, the mice and their nursing mothers were returned to the normal air environment (21% O 2 ) to induce retinal neovascularization in mice. On the 12th day, 14th day, and 16th day after birth, the mice were injected with 1ng, 5ng, and 20ng of IL-37 per gram of mouse body weight, respectively, and the control group was injected with PBS. Among them, mice injected with 20 ng per gram of body weight died. On the 17th day after birth, the mice were intraperitoneally injected...
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