Neutral sea cucumber polysaccharide for resisting type II diabetes mellitus as well as preparation method and application of neutral sea cucumber polysaccharide

Abstract

The invention provides neutral sea cucumber polysaccharide for resisting type II diabetes mellitus as well as a preparation method and application thereof, and the preparation method comprises the following steps: grinding light dry sea cucumber, performing protease enzymolysis, performing anion exchange column separation to obtain sea cucumber polysaccharide, and evaluating the anti-diabetes activity of the sea cucumber polysaccharide through insulin-resistant Hep G2 cells. The activity and activity of the sea cucumber neutral polysaccharide obtained by the method are reported for the first time, the glucose uptake capability of insulin-resistant Hep G2 cells can be remarkably improved, and due to the starch-like structure, the sea cucumber neutral polysaccharide can be used as an active substance and an auxiliary stabilizer in the preparation tabletting process.

Description

technical field

[0001] The invention relates to the field of medical care and health care, and is also a comprehensive utilization of sea cucumber, in particular to a neutral sea cucumber polysaccharide with anti-type II diabetes, a preparation method and an application thereof. Background technique

[0002] Diabetes is a chronic endocrine and metabolic disease characterized by long-term hyperglycemia. It is mainly divided into two types: type I and type II. Type I diabetes is mainly manifested by the lack and deficiency of insulin, but type II diabetes is more complex. It is generally manifested as insulin resistance, that is, the responsiveness of insulin target organs to insulin is reduced, and a series of complications caused by sugar and fat metabolism disorders are a serious threat to human health. The third leading disabling and fatal disease after cancer. Therefore, improving insulin sensitivity and improving insulin resistance in diabetic patients is of great signi...

Examples

Embodiment 1

[0023] A kind of preparation and application of anti-type II diabetes neutral sea cucumber polysaccharide, the specific process parameters are as follows:

[0024] (1) Take 500 g of dried sea cucumber, grind it, add 5 g papain, mix well with 2500 mL of water, enzymolysis at 50°C for 20 h, boil for 20 min to kill the enzyme, adjust pH=2.5, centrifuge to obtain the supernatant, add 7500 The crude polysaccharide was obtained by centrifugation and lyophilization in 2 M KAc ethanol solution at 4 °C overnight, which was eluted with a DEAE anion exchange column with 0 M, 2 M NaCl aqueous solution, and the water elution fraction was dialyzed and lyophilized;

[0025] (2) The sea cucumber polysaccharide was degraded by 2M trifluoroacetic acid at 110℃ for 6h, and the monosaccharide composition was detected by PMP-HPLC; KBr was pressed into tablets and then analyzed by FTIR, and the hydrogen and carbon spectra were obtained by 400 M NMR after heavy water exchange. The results are attache...

Embodiment 2

[0027] A preparation and application of neutral sea cucumber polysaccharide against type II diabetes, and the method for measuring its anti-diabetic activity is as follows:

[0028] (1) The anti-sugar activity of sea cucumber polysaccharide was detected by Hep G2 cells of liver cancer. 5 After plating at a concentration of 1 cells / mL, 18 mM glucosamine was used to induce insulin resistance for 24 hours, sea cucumber polysaccharide samples were added, and rosiglitazone was used as a positive control to detect the inhibitory effect of sea cucumber polysaccharides on insulin resistance of liver cancer cells;

[0029] (2) The cell culture supernatant was collected, and the glucose content was measured by the glucose oxidase method to calculate the glucose consumption of each experimental group. At the same time, the effect of each concentration of sea cucumber polysaccharide on the cell viability was detected by the MTT method. The results are attached figure 2 ;

Embodiment 3

[0031] A kind of preparation and application of anti-type II diabetes neutral sea cucumber polysaccharide, its anti-diabetic tableting formula and preparation are as follows:

[0032] Take 100 g of sea cucumber polysaccharide prepared above, 80 g of Lycium barbarum polysaccharide extract, 20 g of Luo Han Guo saponin extract, 100 g of microcrystalline cellulose, 50 g of xylitol, and 5 g of magnesium stearate, mix them in a mixer, and press into tablets, 200 mg each. piece.

[0033] The above-mentioned three specific embodiments are exemplary, and the purpose is to enable the skilled person to understand this patent, and any equivalent changes or modifications made on the basis of this patent are all within the scope of this patent.

technical field

[0001] The invention relates to the field of medical care and health care, and is also a comprehensive utilization of sea cucumber, in particular to a neutral sea cucumber polysaccharide with anti-type II diabetes, a preparation method and an application thereof. Background technique

[0002] Diabetes is a chronic endocrine and metabolic disease characterized by long-term hyperglycemia. It is mainly divided into two types: type I and type II. Type I diabetes is mainly manifested by the lack and deficiency of insulin, but type II diabetes is more complex. It is generally manifested as insulin resistance, that is, the responsiveness of insulin target organs to insulin is reduced, and a series of complications caused by sugar and fat metabolism disorders are a serious threat to human health. The third leading disabling and fatal disease after cancer. Therefore, improving insulin sensitivity and improving insulin resistance in diabetic patients is of great signi...

Examples

Embodiment 1

[0023] A kind of preparation and application of anti-type II diabetes neutral sea cucumber polysaccharide, the specific process parameters are as follows:

[0024] (1) Take 500 g of dried sea cucumber, grind it, add 5 g papain, mix well with 2500 mL of water, enzymolysis at 50°C for 20 h, boil for 20 min to kill the enzyme, adjust pH=2.5, centrifuge to obtain the supernatant, add 7500 The crude polysaccharide was obtained by centrifugation and lyophilization in 2 M KAc ethanol solution at 4 °C overnight, which was eluted with a DEAE anion exchange column with 0 M, 2 M NaCl aqueous solution, and the water elution fraction was dialyzed and lyophilized;

[0025] (2) The sea cucumber polysaccharide was degraded by 2M trifluoroacetic acid at 110℃ for 6h, and the monosaccharide composition was detected by PMP-HPLC; KBr was pressed into tablets and then analyzed by FTIR, and the hydrogen and carbon spectra were obtained by 400 M NMR after heavy water exchange. The results are attache...

Embodiment 2

[0027] A preparation and application of neutral sea cucumber polysaccharide against type II diabetes, and the method for measuring its anti-diabetic activity is as follows:

[0028] (1) The anti-sugar activity of sea cucumber polysaccharide was detected by Hep G2 cells of liver cancer. 5 After plating at a concentration of 1 cells / mL, 18 mM glucosamine was used to induce insulin resistance for 24 hours, sea cucumber polysaccharide samples were added, and rosiglitazone was used as a positive control to detect the inhibitory effect of sea cucumber polysaccharides on insulin resistance of liver cancer cells;

[0029] (2) The cell culture supernatant was collected, and the glucose content was measured by the glucose oxidase method to calculate the glucose consumption of each experimental group. At the same time, the effect of each concentration of sea cucumber polysaccharide on the cell viability was detected by the MTT method. The results are attached figure 2 ;

Embodiment 3

[0031] A kind of preparation and application of anti-type II diabetes neutral sea cucumber polysaccharide, its anti-diabetic tableting formula and preparation are as follows:

[0032] Take 100 g of sea cucumber polysaccharide prepared above, 80 g of Lycium barbarum polysaccharide extract, 20 g of Luo Han Guo saponin extract, 100 g of microcrystalline cellulose, 50 g of xylitol, and 5 g of magnesium stearate, mix them in a mixer, and press into tablets, 200 mg each. piece.

[0033] The above-mentioned three specific embodiments are exemplary, and the purpose is to enable the skilled person to understand this patent, and any equivalent changes or modifications made on the basis of this patent are all within the scope of this patent.

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