Application of oligochitosan in preparing medicine for preventing from formation of atherosclerosis

Abstract

The invention relates to a medicine for curing atherosclerosis, in particular to application of oligochitosan in preparing a medicine for preventing from the formation of atherosclerosis. The application takes lipopolysaccharide (lipopolysaccharide) and humanized gene recombinant tumor necrosis factor-alpha (TNF-alpha) as an induction factor, and takes human umbilical vein endothelial cell as a research object, thereby having the following results: (1) the oligochitosan can obviously restrain the expressions of IL-8, IL-6 and relevant inflammatory factors in the lipopolysaccharide-induced human umbilical vein endothelial cell; (2) the oligochitosan can obviously restrain the expressions of membrane surface molecules VCAM-1. ICAM-1 and relevant adhesion molecules family members of the TNF-alpha-induced human umbilical vein endothelial cell; and (3) the oligochitosan can obviously restrain the adhesion between the TNF-alpha-induced human umbilical vein endothelial cell and the human mononuclear cell.

Description

technical field

[0001] The invention relates to a medicine for treating atherosclerosis, specifically the application of chitosan oligosaccharides in the preparation of medicines for preventing the formation of atherosclerosis, and further to the application of chitosan oligosaccharides in the preparation of arteriosclerosis caused by bacterial infection and inflammatory factors. Application of atherosclerotic medicine or health food. Background technique

[0002] Atherosclerosis refers to a combination of lesions caused by focal deposition of lipids, carbohydrates, blood components, connective tissue, and calcium, accompanied by changes in the arterial media. So far, atherosclerosis has become a major disease that threatens the safety of human life in western developed countries. With the improvement of living standards and changes in eating habits in our country, atherosclerosis has also become an important reason affecting the health of Chinese people. According to the ...

Examples

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 ...

technical field

[0001] The invention relates to a medicine for treating atherosclerosis, specifically the application of chitosan oligosaccharides in the preparation of medicines for preventing the formation of atherosclerosis, and further to the application of chitosan oligosaccharides in the preparation of arteriosclerosis caused by bacterial infection and inflammatory factors. Application of atherosclerotic medicine or health food. Background technique

[0002] Atherosclerosis refers to a combination of lesions caused by focal deposition of lipids, carbohydrates, blood components, connective tissue, and calcium, accompanied by changes in the arterial media. So far, atherosclerosis has become a major disease that threatens the safety of human life in western developed countries. With the improvement of living standards and changes in eating habits in our country, atherosclerosis has also become an important reason affecting the health of Chinese people. According to the ...

Examples

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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