Method for preparing functional beverage for treating rheumatoid arthritis

A technology for rheumatoid arthritis and beverages, applied in the biological field, can solve the problems of large side effects and low efficacy of drugs, and achieve the effect of shortening the treatment time

Inactive Publication Date: 2017-09-08

UNIV OF JINAN

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AI-Extracted Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to provide a drink for treating rheumatoid arthritis in...

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Abstract

The invention discloses a method for preparing a functional beverage for treating rheumatoid arthritis, and belongs to the technical field of biology. The functional beverage for treating rheumatoid arthritis is prepared by compounding a bee venom bioactive peptide solution and a propolis flavone solution, wherein the bee venom bioactive peptide solution is an animal polypeptide solution separated from bee venom, and the propolis flavone solution is an active substance extracted from raw propolis. A sephadex column chromatography is combined with high-speed crushing, centrifugation and an organic solvent extraction method to obtain bee venom bioactive peptide and propolis flavone. Compared with drugs namely aspirin, the beverage for treating rheumatoid arthritis has the effect of treating the rheumatoid arthritis without damaging renal functions. After the beverage provided by the invention is drunk, pains of bodies suffering from rheumatism can be relieved to a certain extent, and the beverage is small in side effects on bodies, and can be drunk for a long term.

Application Domain

Dispersion deliveryPeptide/protein ingredients +7

Technology Topic

Side effectOrganic solvent +16

Examples

Experimental program(3)

Example Embodiment

[0027] Example 1:

[0028] ① Preparation of propolis flavone solution

[0029] Put the propolis into a white wine with an alcohol content of 60°, extract it in a water bath at 80°C for 3 hours, and extract once. Centrifuge at 5000r/min for 5min for solid-liquid separation to remove solid propolis residues. Use a white wine with an alcohol content of 60° to remove the propolis The flavone extract is configured into a propolis flavone solution with a concentration of 0.326 mg/mL; the alcohol content is the volume percentage of ethanol contained in the food-grade liquor.

[0030] ② Preparation of bee venom active peptide solution

[0031] The bee venom was eluted with distilled water through Sephadex G-25, the flow rate was controlled at 28mL/h, the molecular weight of each separated component was determined, and the layers with molecular weights of 2035Da, 2587Da and 2840Da were collected under the Sephadex column. Analyze the components and freeze-dry, the obtained solid sample is the meli venom active peptide, and the meli venom active peptide is prepared with purified water into a meli venom active peptide solution with a concentration of 0.05 mg/mL;

[0032] The dextran gel G-25 is a dextran represented by the English letter G, and G reflects the degree of crosslinking, swelling and distribution of the gel, and 25 means 2.5 grams of water per gram of gel when it swells;

[0033] The elution is using distilled water as the mobile phase to continuously pass through the stationary phase of the chromatographic column, the mobile phase and the stationary phase are weaker than the sample, and the components of the sample are washed out from the stationary phase in sequence;

[0034] The Da is the unit of the molecular weight of the active peptide of bee venom;

[0035] The determination of the molecular weight is based on the standard curve regression equation of the standard elution retention time T (min) and its relative molecular weight logarithmic value (lgMr). The standard is an antimicrobial peptide standard (Mr =1422Da), oxidized glutathione Standard product (Mr =612Da), reduced glutathione standard product (Mr =307.32Da).

[0036] The freeze-drying is a drying method in which the obtained components are frozen to below the freezing point of water and placed in a high-vacuum (10-40Pa) container, and the water in the material is directly sublimated from solid ice to vapor by heating ;

[0037] The melittin active peptides include melittin, melittin, and degranulation peptides for cell hypertrophy;

[0038] ③Preparation of drinks for treating rheumatoid arthritis

[0039] Mix ① the 0.05mg/mL bee venom active peptide solution with the ② 0.326mg/mL propolis flavone solution according to the volume ratio V (bee venom active peptide): V (propolis) = 1:7 , The result is a drink with the function of treating rheumatoid arthritis.

[0040] The volume ratio is the ratio of the volume of the bee venom active peptide solution to the volume of the propolis flavone solution;

[0041] The compound is a mixture of bee venom active peptide solution and propolis flavone solution.

[0042] Normal mice TNF-α, IL-6, IL-1β and blood urine nitrogen values were 99.29±5.89 pg/mL, 100.81±8.72 pg/mL, 101.29±7.39 pg/mL, 126.64±32.06 mmol/L, and no drinking The TNF-α, IL-6, IL-1β and blood urine nitrogen values of the mixed beverage mice were 110.54±5.64 pg/mL, 216.22±12.86 pg/mL, 110.35±5.49 pg/mL, and 187.17±8.54 mmol/L, respectively. The TNF-α, IL-6, IL-1β and blood urine nitrogen values of mice who drank the compound drink were 100.25±5.82pg/mL, 106.04±9.76pg/mL, 107.23±9.55 pg/mL, 151.53±54.99 mmol/L, respectively , TNF-α, IL-6, IL-1β and blood urine nitrogen values of mice taking aspirin were 106.37±2.38 pg/mL, 116.87±13.34 pg/mL, 102.55±4.88 pg/mL, and 156.63±33.89 mmol/L, respectively.

Example Embodiment

[0043] Example 2:

[0044] ① Preparation of propolis flavone solution

[0045] Put the propolis into a white wine with an alcohol content of 70°, extract in a water bath at 90°C for 4 hours, and extract 2 times. Centrifuge at 4000r/min for 6min for solid-liquid separation to remove solid propolis residues. Use a white wine with an alcohol content of 70° to remove the propolis. The flavone extract is configured into a propolis flavone solution with a concentration of 0.217mg/mL;

[0046] The alcohol content is the volume percentage of ethanol contained in food-grade liquor.

[0047] ② Preparation of bee venom active peptide solution

[0048] The bee venom was eluted with distilled water through Sephadex G-25, the flow rate was controlled at 30mL/h, the molecular weight of each separated component was determined, and the layers with molecular weights of 2035Da, 2587Da and 2840Da were collected under the Sephadex column. The components are analyzed and freeze-dried, the obtained solid sample is the meli venom active peptide, and the meli venom active peptide is prepared with purified water into a meli venom active peptide solution with a concentration of 0.03 mg/mL;

[0049] The dextran gel G-25 is a dextran represented by the English letter G, and G reflects the degree of crosslinking, swelling and distribution of the gel, and 25 means 2.5 grams of water per gram of gel when it swells;

[0050] The elution is using distilled water as the mobile phase to continuously pass through the stationary phase of the chromatographic column, the mobile phase and the stationary phase are weaker than the sample, and the components of the sample are washed out from the stationary phase in sequence;

[0051] The Da is the unit of the molecular weight of the active peptide of bee venom;

[0052] The determination of the molecular weight is based on the standard curve regression equation of the standard elution retention time T (min) and its relative molecular weight logarithmic value (lgMr). The standard is an antimicrobial peptide standard (Mr =1422Da), oxidized glutathione Standard product (Mr =612Da), reduced glutathione standard product (Mr =307.32Da).

[0053] The freeze-drying is a drying method in which the obtained components are frozen to below the freezing point of water and placed in a high-vacuum (10-40Pa) container, and the water in the material is directly sublimated from solid ice to vapor by heating ;

[0054] The melittin active peptides include melittin, melittin, and degranulation peptides for cell hypertrophy;

[0055] ③Preparation of drinks for treating rheumatoid arthritis

[0056] Mix the propolis active peptide solution with a concentration of 0.03mg/mL in ① and the propolis flavone solution with a concentration of 0.217mg/mL in ② according to the volume ratio V (bee venom active peptide): V (propolis) = 1:9 , The result is a drink with the function of treating rheumatoid arthritis.

[0057] The volume ratio is the ratio of the volume of the bee venom active peptide solution to the volume of the propolis flavone solution;

[0058] The compound is a mixture of bee venom active peptide solution and propolis flavone solution.

[0059] Normal mice TNF-α, IL-6, IL-1β and blood urine nitrogen values were 99.29±5.89 pg/mL, 100.81±8.72 pg/mL, 101.29±7.39 pg/mL, 126.64±32.06 mmol/L, and do not drink The TNF-α, IL-6, IL-1β and blood urine nitrogen values of the mixed beverage mice were 110.54±5.64 pg/mL, 216.22±12.86 pg/mL, 110.35±5.49 pg/mL, and 187.17±8.54 mmol/L, respectively. The TNF-α, IL-6, IL-1β and blood urine nitrogen values of the mice drinking the compound drink were 100.91±7.89pg/mL, 109.87±3.22pg/mL, 103.64±4.95pg/mL, 189.94±27.17mmol/L, respectively The TNF-α, IL-6, IL-1β and blood urine nitrogen values of mice taking aspirin were 106.37±2.38 pg/mL, 116.87±13.34 pg/mL, 102.55±4.88 pg/mL, and 156.63±33.89 mmol/L, respectively.

Example Embodiment

[0060] Example 3:

[0061] ① Preparation of propolis flavone solution

[0062] Put the propolis into a liquor with an alcohol content of 50°, extract in a water bath at 70°C for 2 hours, and extract 3 times. Centrifuge at 3000r/min for 4min for solid-liquid separation to remove solid propolis residues. The flavonoid extract is configured into a propolis solution with a concentration of 0.109mg/mL;

[0063] The alcohol content is the volume percentage of ethanol contained in food-grade liquor.

[0064] ② Preparation of bee venom active peptide solution

[0065] The bee venom was eluted with distilled water through Sephadex G-25, the flow rate was controlled at 26mL/h, the molecular weight of each separated component was determined, and the layers with molecular weights of 2035Da, 2587Da and 2840Da were collected under the Sephadex column. The components are analyzed and freeze-dried. The obtained solid sample is the meli venom active peptide. The meli venom active peptide is prepared with purified water to form a meli venom active peptide solution with a concentration of 0.0182 mg/mL;

[0066] The dextran gel G-25 is a dextran represented by the English letter G, and G reflects the degree of crosslinking, swelling and distribution of the gel, and 25 means 2.5 grams of water per gram of gel when it swells;

[0067] The elution is using distilled water as the mobile phase to continuously pass through the stationary phase of the chromatographic column, the mobile phase and the stationary phase are weaker than the sample, and the components of the sample are washed out from the stationary phase in sequence;

[0068] The Da is the unit of the molecular weight of the active peptide of bee venom;

[0069] The determination of the molecular weight is based on the standard curve regression equation of the standard elution retention time T (min) and its relative molecular weight logarithmic value (lgMr). The standard is an antimicrobial peptide standard (Mr =1422Da), oxidized glutathione Standard product (Mr =612Da), reduced glutathione standard product (Mr =307.32Da).

[0070] The freeze-drying is a drying method in which the obtained components are frozen to below the freezing point of water and placed in a high-vacuum (10-40Pa) container, and the water in the material is directly sublimated from solid ice to vapor by heating ;

[0071] The melittin active peptides include melittin, melittin, and degranulation peptides for cell hypertrophy;

[0072] ③Preparation of drinks for treating rheumatoid arthritis

[0073] Mix the propolis active peptide solution with a concentration of 0.0182mg/mL in ① and the propolis flavone solution with a concentration of 0.109mg/mL in ② according to the volume ratio V (bee venom active peptide): V (propolis) = 1:5 , The result is a drink with the function of treating rheumatoid arthritis.

[0074] The volume ratio is the ratio of the volume of the bee venom active peptide solution to the volume of the propolis flavone solution;

[0075] The compound is a mixture of bee venom active peptide solution and propolis flavone solution.

[0076] Normal mice TNF-α, IL-6, IL-1β and blood urine nitrogen values were 99.29±5.89 pg/mL, 100.81±8.72 pg/mL, 101.29±7.39 pg/mL, 126.64±32.06 mmol/L, and do not drink The TNF-α, IL-6, IL-1β and blood urine nitrogen values of the mixed beverage mice were 110.54±5.64 pg/mL, 216.22±12.86 pg/mL, 110.35±5.49 pg/mL, 187.17±8.54 mmol/L, respectively. The TNF-α, IL-6, IL-1β and blood urine nitrogen values of the mice who drank the mixed drink were 100.70±18.46pg/mL, 107.89±5.39pg/mL, 109.21±2.46pg/mL, 209.99±12.28mmol/L, respectively , The TNF-α, IL-6, IL-1β and blood urine nitrogen values of mice taking aspirin were 106.37±2.38 pg/mL, 116.87±13.34 pg/mL, 102.55±4.88 pg/mL, and 156.63±33.89 mmol/L, respectively.

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Description & Claims & Application Information

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Method for preparing functional beverage for treating rheumatoid arthritis

A technology for rheumatoid arthritis and beverages, applied in the biological field, can solve the problems of large side effects and low efficacy of drugs, and achieve the effect of shortening the treatment time

AI-Extracted Technical Summary

Problems solved by technology

Abstract

Examples

Example Embodiment

Example Embodiment

Example Embodiment

PUM

Description & Claims & Application Information

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