Application of oligochitosan in preparing medicine for preventing from formation of atherosclerosis
A technology of atherosclerosis and chitosan oligosaccharides, applied in the direction of drug combinations, pharmaceutical formulas, organic active ingredients, etc., to achieve the effects of inhibiting production, regulating blood lipids, and improving the body's immunity
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Embodiment 1
[0028] Oligochitosan is administered in the form of an aqueous solution, and the specific operation of the liquid preparation is as follows: ① Accurately weigh 5 mg of oligochitosan, dissolve it in 5 ml of DMEM-F12 medium containing 1% fetal bovine serum, and form a final concentration of 1 mg / ml of chitosan oligosaccharide solution; ②Use a 0.22μM pore size needle filter to filter the aforementioned drug solution, which is a sterile chitosan oligosaccharide stock solution; ③After aliquoting the above stock solution, store Saved for use in the following examples.
Embodiment 2
[0030] Analysis of transcriptional inhibition of IL-8 and IL-6 in human umbilical vein endothelial cells by oligochitosan. The main operations are as follows: ① The experiment is divided into 5 groups ( figure 1 ), that is, normal cell culture group (Control), lipopolysaccharide model group, chitooligosaccharide preprotected group 1 (50 μg / ml), chitooligosaccharide preprotected group 2 (100 μg / ml) and chitooligosaccharide preprotected group 3 ( 200 μg / ml). ②Digest monolayer cultured human umbilical vein endothelial cells with 0.125% trypsin, and press 2×10 6 Each bottle is inoculated in a 25ml culture flask, and the total volume of cell suspension added to each bottle is 4ml. ③Put the culture bottle into the cell culture incubator, 5% CO 2 , 37°C, cultivated for 24h. ④Take out the culture bottle and discard the supernatant, add 3ml of fresh DMEM-F12 medium (Gibco, Grand Island, USA) to the Control group and lipopolysaccharide group respectively; add 50, 100 and 200 μg / ml...
Embodiment 3
[0033] Inhibitory analysis of chitooligosaccharides on IL-8 and IL-6 protein expression in human umbilical vein endothelial cells. The main operations are as follows: ① The experiment is divided into 5 groups ( figure 2 ), that is, normal cell culture group (Control), lipopolysaccharide model group, chitooligosaccharide preprotected group 1 (50 μg / ml), chitooligosaccharide preprotected group 2 (100 μg / ml) and chitooligosaccharide preprotected group 3 ( 200 μg / ml). ②Digest monolayer cultured human umbilical vein endothelial cells with 0.125% trypsin, and press 5×10 3 Density per well Cells were seeded in 96 wells, 5 wells were replicated in each group, and the volume of cell suspension added to each well was 150 μl. ③ Put the culture plate into the cell culture incubator, 5% CO 2 , 37°C, cultivated for 24h. ④Take out the culture plate and discard the supernatant, add 150 μl of fresh DMEM-F12 medium to the Control group and lipopolysaccharide group respectively; add 150 μl ...
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