Use of cortex eucommiae total polyoses in the preparing of the medicine for preventing and treating systematic lupus erythematosus

Abstract

The invention belong to Chinese traditional medicine field, relates to the application of eucommia polysaccharide for preparing medicine preventing systematic Cazenave's lupus. The invention separates polysaccharide extract from eucommia, the average productivity is 2.2%, and the polysaccharide content is more than 68%. It is showed by whole animal model test that the protective effect for hyper reaction and kidney injury for mice by said polysaccharide is distinctive, and it is proven that polysaccharide can prevent Cazenave's lupus syndrome. The eucommia polysaccharide can also be used to produce medicine preventing and treating self-immune disease.

Description

Technical field

[0001] The invention belongs to the field of traditional Chinese medicine, and relates to a new medicinal use of the total polysaccharide of the traditional Chinese medicine Eucommia ulmoides. Specifically, it relates to the use of Eucommia ulmoides total polysaccharides in the preparation of systemic lupus erythematosus drugs. Background technique:

[0002] Systemic lupus erythematosus (SLE) is an inflammatory connective tissue disease of unknown etiology. It mainly affects women and can also affect children. A multi-country and multi-center study showed that the current incidence of systemic lupus erythematosus is 12.5 / 100,000 to 39 / 100,000. SLE is an autoimmune disease involving multiple systems, complicated clinical manifestations, protracted and repeated course, and high fatality rate. It is usually regarded as a representative autoimmune disease. The phenotypes of SLE autoimmune abnormalities are highly diverse, including immune tolerance defects, lymphocyte...

Examples

Embodiment 1

[0038] Take 100g of Eucommia medicinal materials, crush them, and extract them with 95% ethanol cold soaking for three times, each time for 7 days, filter, and dry the medicine residue in a ventilated place at room temperature, then extract 3 times with hot water, filter, combine the extracts, concentrate, Centrifuge the supernatant with trichloroacetic acid to remove free protein, centrifuge, concentrate the supernatant and dialyze it in running water for 3 days, concentrate the dialysate to a small volume and add ethanol to the alcohol content of 80%, wash the precipitate with acetone, ethanol and freeze-dry The total polysaccharide extract is obtained with a yield of more than 2% and a polysaccharide content of more than 68%.

Embodiment 2

[0040]Eucommia (Eucomia ulmoides Olive.) total polysaccharide 3.0mg.ml -1 Use barbital buffer (VBS2+) to double-dilute solutions with eight concentrations. Take 0.2ml samples of each concentration and add 0.2ml complement (guinea pig serum 1:32 dilution), 0.1ml 2% sheep red blood cells (sheep red blood). cell, SRBC) and 0.1ml 1:1000 hemolysin (anti-SRBC serum). Put each tube into a low-temperature high-speed centrifuge at 2500G and centrifuge at 4°C after 30 minutes in a 37°C water bath. After 20 minutes, take 0.25ml of each tube’s supernatant in Measure the absorbance at 405nm with a microplate reader. At the same time, 0.2ml of each concentration sample solution was mixed with 0.4ml VBS2+, placed in a low-temperature high-speed centrifuge, centrifuged at 2500G, 4℃ for 10min, and 0.25ml of supernatant from each tube was measured at 405nm with a microplate reader as the sample absorbance color value. Then subtract the absorbance of the sample at the corresponding concentration fro...

Embodiment 3

[0042] Eucommia (Eucomia ulmoides Olive.) total polysaccharide 3.0mg.ml -1 Use EGTA-containing barbital buffer (EGTA-VBS) to double-dilute a solution of eight concentrations. Take 0.15ml samples of each concentration and add 0.15ml complement (human serum 1:10 dilution), 0.2ml 2% rabbit red blood cells , Put each tube into a low-temperature high-speed centrifuge 25006 at 37℃ for 30 minutes and centrifuge at 4℃. After 20 minutes, take 0.25ml of supernatant from each tube and measure the absorbance at 405nm in a microplate reader. Then mix 0.15ml of each concentration sample solution with 0.35ml EGTA-VBS, put it in a low-temperature high-speed centrifuge and centrifuge at 2500G, 4℃ for 10 minutes, take 0.25ml of supernatant from each tube and measure the absorbance at 405nm in the microplate reader, as the sample absorbance color value . Then subtract the absorbance of the sample at the corresponding concentration from the absorbance measured by the titer of each sample at different...

Technical field

[0001] The invention belongs to the field of traditional Chinese medicine, and relates to a new medicinal use of the total polysaccharide of the traditional Chinese medicine Eucommia ulmoides. Specifically, it relates to the use of Eucommia ulmoides total polysaccharides in the preparation of systemic lupus erythematosus drugs. Background technique:

[0002] Systemic lupus erythematosus (SLE) is an inflammatory connective tissue disease of unknown etiology. It mainly affects women and can also affect children. A multi-country and multi-center study showed that the current incidence of systemic lupus erythematosus is 12.5 / 100,000 to 39 / 100,000. SLE is an autoimmune disease involving multiple systems, complicated clinical manifestations, protracted and repeated course, and high fatality rate. It is usually regarded as a representative autoimmune disease. The phenotypes of SLE autoimmune abnormalities are highly diverse, including immune tolerance defects, lymphocyte...

Examples

Embodiment 1

[0038] Take 100g of Eucommia medicinal materials, crush them, and extract them with 95% ethanol cold soaking for three times, each time for 7 days, filter, and dry the medicine residue in a ventilated place at room temperature, then extract 3 times with hot water, filter, combine the extracts, concentrate, Centrifuge the supernatant with trichloroacetic acid to remove free protein, centrifuge, concentrate the supernatant and dialyze it in running water for 3 days, concentrate the dialysate to a small volume and add ethanol to the alcohol content of 80%, wash the precipitate with acetone, ethanol and freeze-dry The total polysaccharide extract is obtained with a yield of more than 2% and a polysaccharide content of more than 68%.

Embodiment 2

[0040]Eucommia (Eucomia ulmoides Olive.) total polysaccharide 3.0mg.ml -1 Use barbital buffer (VBS2+) to double-dilute solutions with eight concentrations. Take 0.2ml samples of each concentration and add 0.2ml complement (guinea pig serum 1:32 dilution), 0.1ml 2% sheep red blood cells (sheep red blood). cell, SRBC) and 0.1ml 1:1000 hemolysin (anti-SRBC serum). Put each tube into a low-temperature high-speed centrifuge at 2500G and centrifuge at 4°C after 30 minutes in a 37°C water bath. After 20 minutes, take 0.25ml of each tube’s supernatant in Measure the absorbance at 405nm with a microplate reader. At the same time, 0.2ml of each concentration sample solution was mixed with 0.4ml VBS2+, placed in a low-temperature high-speed centrifuge, centrifuged at 2500G, 4℃ for 10min, and 0.25ml of supernatant from each tube was measured at 405nm with a microplate reader as the sample absorbance color value. Then subtract the absorbance of the sample at the corresponding concentration fro...

Embodiment 3

[0042] Eucommia (Eucomia ulmoides Olive.) total polysaccharide 3.0mg.ml -1 Use EGTA-containing barbital buffer (EGTA-VBS) to double-dilute a solution of eight concentrations. Take 0.15ml samples of each concentration and add 0.15ml complement (human serum 1:10 dilution), 0.2ml 2% rabbit red blood cells , Put each tube into a low-temperature high-speed centrifuge 25006 at 37℃ for 30 minutes and centrifuge at 4℃. After 20 minutes, take 0.25ml of supernatant from each tube and measure the absorbance at 405nm in a microplate reader. Then mix 0.15ml of each concentration sample solution with 0.35ml EGTA-VBS, put it in a low-temperature high-speed centrifuge and centrifuge at 2500G, 4℃ for 10 minutes, take 0.25ml of supernatant from each tube and measure the absorbance at 405nm in the microplate reader, as the sample absorbance color value . Then subtract the absorbance of the sample at the corresponding concentration from the absorbance measured by the titer of each sample at different...

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