CN115736255A - Application of high molecular weight oat beta-glucan extract - Google Patents
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Abstract
The invention discloses a high molecular weight oat beta-glucan extract, which is characterized in that the content of beta-glucan with the relative molecular mass of more than or equal to 300kDa is more than or equal to 95 percent, and the extract has the function of relieving abnormal glycolipid metabolism induced by food-borne advanced glycation end products. The high molecular weight oat beta-glucan extract is obtained by pretreatment, extraction, concentration, separation, adsorption, nanofiltration, drying and determination; the food-borne advanced glycation end product can obviously reduce 19-25% of body weight and 8-21% of blood sugar of a model animal induced by the food-borne advanced glycation end product, obviously relieve 9-17% of oral glucose tolerance damage of the model animal, obviously inhibit insulin resistance, and obviously inhibit fat accumulation in abdominal fat tissue, perirenal fat tissue, epididymal fat tissue and mesenteric fat tissue of the model animal. The oat beta-glucan extract has high content of high molecular weight beta-glucan, and can be used for preparing food, feed, human or veterinary drugs.
Description
Technical Field
The invention belongs to the technical field of biological resource utilization, and particularly relates to application of a high molecular weight oat beta-glucan extract.
Background
Numerous studies have demonstrated that food-borne advanced glycation end products (AGEs) can have a non-negligible effect on body glycolipid metabolism and energy homeostasis, and that this effect directly/indirectly contributes to the global pandemic of obesity and diabetes in the last few decades. Therefore, targeted dietary intervention is necessary to alleviate/eliminate glycolipid metabolism disorders associated with diets high in AGEs. Oat beta-glucan (Oat beta-glucan) is a natural soluble dietary fiber composed of glucose molecules linked together by a series of beta- (1-3) and beta- (1-4) linkages. The soluble dietary fiber can enhance the activity of macrophages, quickly kill invading pathogenic microorganisms such as viruses, bacteria and fungi, improve immunity, regulate blood sugar, effectively delay postprandial blood sugar rise, protect intestinal tracts, prevent diarrhea or constipation, reduce cholesterol, resist cancer and radiation, and has wide reports of beneficial effects on patients with diabetes, hypertension and cardiovascular diseases. The high molecular weight soluble dietary fiber has the potential of regulating blood fat; the European guidelines for dyslipidemia management indicate that a daily intake of 5-15 g of high molecular weight soluble dietary fiber reduces blood total cholesterol and low density lipoprotein levels; the U.S. guidelines recommend a daily intake of 10 to 25g of high molecular weight soluble dietary fiber. In addition, high molecular weight oat beta-glucan as a soluble dietary fiber can form a viscous layer in the small intestine interfering with the absorption of dietary small molecules. In addition, the high molecular weight oat beta-glucan can generate short-chain fatty acid through intestinal flora fermentation, promote the proliferation of beneficial bacteria and inhibit the proliferation of harmful bacteria. The existing extraction method of oat beta-glucan mainly comprises the steps of treating and degrading starch by using thermostable alpha-amylase, and then adding ammonium sulfate to primarily purify the extracted beta-glucan. The method introduces a large amount of small molecule free sugar, protein and some nonpolar compounds, and large amount of edible sugar is not beneficial to glycolipid metabolism and energy steady-state stabilization of organism.
Disclosure of Invention
The invention aims to provide application of a high molecular weight oat beta-glucan extract.
The purpose of the invention is realized by the following technical scheme:
the application of the high molecular weight oat beta-glucan extract in relieving glycolipid metabolic abnormality induced by food-borne advanced glycation end products is characterized in that the relative molecular mass of the oat beta-glucan extract is more than or equal to 300kDa, the purity of the oat beta-glucan extract is more than or equal to 95 percent.
Preferably, the high molecular weight oat β -glucan extract is prepared by:
(1) Extracting oat powder with water at 70-90 ℃, filtering, concentrating and centrifuging the extract to obtain supernatant;
(2) Performing column chromatography on the supernatant obtained in the step (1) to obtain a colorless oat extract;
(3) And (3) taking the colorless oat extract obtained in the step (2), performing pressure filtration through a 300kDa nanofiltration tube, and drying to obtain the high-molecular-weight oat beta-glucan extract.
Preferably, the column chromatography in step (2) is a supramolecular gel column NBP5.
Preferably, the mass ratio of the oat powder to the water in the step (1) is 1.
Preferably, the filtrate is concentrated to 1/3-1/4 of the original volume in the step (1); centrifuging for 5-10 min at the rotating speed of 2500-3000 Xg.
Preferably, the oat flour in step (1) is obtained by the following steps: the oat is cleaned, aired, crushed and sieved by a 60-mesh sieve to obtain oat powder.
Application of high molecular weight oat beta-glucan extract in preparation of food, feed and medicaments for preventing and/or relieving abnormal glycolipid metabolism induced by food-borne advanced glycation end products.
A food product having the effect of preventing and/or alleviating glycolipid metabolic abnormalities induced by food-borne advanced glycation end products, said food product comprising a high molecular weight oat β -glucan extract, said high molecular weight oat β -glucan extract being ingested in an amount of 0.3 to 5 g/day.
A feed for preventing and/or alleviating glycolipid metabolism abnormality induced by food-borne late-stage saccharification end products, which comprises a high molecular weight oat beta-glucan extract, wherein the addition amount of the high molecular weight oat beta-glucan extract in the feed is 1-10% of the dry weight of the feed.
A medicament having the effect of preventing and/or alleviating glycolipid metabolic abnormality induced by food-borne advanced glycation end products, which comprises a high molecular weight oat beta-glucan extract, wherein the content of the high molecular weight oat beta-glucan extract is 20-50%.
Compared with the prior art, the invention has the following advantages:
the high molecular weight oat beta-glucan extract obtained by the invention can be taken together with the food-derived advanced glycation end product and taken every other day together with the food-derived advanced glycation end product to obviously inhibit the abnormal increase of the body weight and the blood sugar of the glycolipid metabolic abnormality model animal induced by the food-derived advanced glycation end product, obviously relieve the damage of oral glucose tolerance (OGTT), obviously inhibit insulin resistance and obviously inhibit fat accumulation in abdominal cavity adipose tissue, perirenal adipose tissue, epididymal adipose tissue and mesenteric adipose tissue of the glycolipid metabolic abnormality model animal induced by the food-derived advanced glycation end product. Has the effects of preventing and relieving abnormal glycolipid metabolism induced by food-derived advanced glycation end products, and can be used as food, feed or raw materials of human and animal medicines.
Drawings
Figure 1 is the effect of dietary intervention on mouse body weight: fig. 1 (a) is a graph of body weight change of mice in different groups, compared to LA group, indicating P <0.05, indicating P <0.001; FIG. 1 (B) is the average body weight of each group of mice at day 53 of the experiment. Note: * Denotes P <0.001.
Figure 2 is the effect of dietary intervention on blood glucose and glucose tolerance in mice: FIG. 2 (A) is the mean fasting plasma glucose concentration (0 min) for each group of mice at day 54 of the experiment; FIG. 2 (B) is the change in blood glucose concentration in mice in the Oral Glucose Tolerance Test (OGTT); FIG. 2 (C) is the blood glucose concentration of mice at 120min of the Oral Glucose Tolerance Test (OGTT). Note: * P <0.05, P <0.001.
FIG. 3 is a graph of the effect of dietary intervention on fasting plasma insulin and blood glycated hemoglobin levels in mice: FIG. 3 (A) is fasting plasma insulin levels in mice after an 8-week experiment; FIG. 3 (B) is the glycated hemoglobin level of mouse blood after 8 weeks of the experiment. Note: * P <0.05, P <0.01, P <0.001.
FIG. 4 shows the weights of the liver, kidney and adipose tissue of each group of mice: FIG. 4 (A) is the liver weights of the groups of mice after the 8-week trial period; FIG. 4 (B) is a graph of kidney weight of groups of mice after 8 weeks of the trial period; FIG. 4 (C) is the abdominal fat weight of each group of mice after 8 weeks of the test period; FIG. 4 (D) is the perirenal fat weight of each group of mice after 8 weeks of the test period; FIG. 4 (E) is the epididymal fat weight of each group of mice after 8 weeks of the test period; FIG. 4 (F) is the mesenteric fat weight of each group of mice after 8 weeks of the test period. Note: * P <0.05, P <0.01, P <0.001.
Detailed Description
The present invention is further described with reference to the following drawings and examples, but the invention is not limited thereto in any way, and any modifications or alterations made according to the technical teaching and teachings of the present invention are within the scope of the present invention.
The high molecular weight oat beta-glucan extract is obtained by extracting oat, wherein the content of beta-glucan with the relative molecular mass of more than or equal to 300kDa is more than or equal to 95 percent, the high molecular weight oat beta-glucan extract has the function of relieving the glycolipid metabolic abnormality induced by the food-borne advanced glycation end product, and is used for preparing food, feed or medicine for preventing and/or relieving the glycolipid metabolic abnormality induced by the food-borne advanced glycation end product, and the oat beta-glucan extract is prepared by the following steps:
selecting complete fresh oat, cleaning, airing, crushing, and sieving with a 60-mesh sieve to obtain oat powder; weighing oat powder, adding purified water according to a material-liquid ratio of 1-30 to 1; taking the filtrate, and concentrating the filtrate under reduced pressure to 1/3-1/4 of the volume of the filtrate to obtain concentrated solution; placing the supernatant of the concentrated solution in a centrifuge, centrifuging for 5-10 min at the rotating speed of 2500-3000 Xg, and taking the supernatant; slowly pressurizing the supernatant, injecting the supernatant into a column filled with supramolecular gel NBP5, performing column chromatography, and removing high-molecular organic dye in the extract to obtain colorless oat extract; taking colorless oat extract, and performing pressure filtration through a 300kDa nanofiltration tube to remove all components with the relative molecular mass lower than 300kDa in the extract, thereby obtaining high molecular weight oat beta-glucan extract; vacuum freeze drying the high molecular weight oat beta-glucan extracting solution to obtain high molecular weight oat beta-glucan extract powder; verifying the relative molecular mass distribution of beta-glucan in the obtained extract by adopting GPC; the purity of the oat beta-glucan with high molecular weight in the extract is verified by adopting alpha-amylase and beta-glucanase; and determining the content of high molecular weight beta-glucan in the obtained extract by adopting a dye Congo red colorimetric method.
Example 1
Selecting complete fresh oat, cleaning, airing, crushing, and sieving with a 60-mesh sieve to obtain oat powder; weighing oat powder, adding purified water according to a material-liquid ratio of 1, reflux-extracting at 80 ℃ for 80min, filtering, extracting twice, combining filtrates, and standing; concentrating the filtrate under reduced pressure to 1/3 of the filtrate volume to obtain concentrated solution; placing the supernatant in a centrifuge, centrifuging at 3000 Xg for 5min, and collecting supernatant; slowly pressurizing the supernatant, injecting the supernatant into a column filled with supramolecular gel NBP5, performing column chromatography, and removing high-molecular organic dye in the extract to obtain colorless oat extract; taking colorless oat extract, and performing pressure filtration through a 300kDa nanofiltration tube to remove all components with the relative molecular mass lower than 300kDa in the extract, thereby obtaining high molecular weight oat beta-glucan extract; vacuum freeze drying the high molecular weight oat beta-glucan extracting solution to obtain high molecular weight oat beta-glucan extract powder; determining the relative molecular mass of beta-glucan in the obtained extract by adopting GPC (GPC) to be 366.9kDa; the alpha-amylase and the beta-glucanase are adopted to verify that the high molecular weight oat beta-glucan in the extract can be degraded by the beta-glucanase but can not be degraded by the alpha-amylase; the content of high molecular weight beta-glucan in the obtained extract is 96.1 percent by adopting a dye Congo red colorimetric method.
Example 2
Selecting complete fresh oat, cleaning, airing, crushing, and sieving with a 60-mesh sieve to obtain oat powder; weighing oat powder, adding purified water according to a material-liquid ratio of 1 to 40, performing reflux extraction at 80 ℃ for 90min, filtering, extracting twice, combining filtrates, and standing; concentrating the filtrate under reduced pressure to 1/3 of the filtrate volume to obtain concentrated solution; placing the supernatant in a centrifuge, centrifuging at 3000 Xg for 5min, and collecting the supernatant; slowly pressurizing the supernatant, injecting the supernatant into a column filled with supramolecular gel NBP5, performing column chromatography, and removing high-molecular organic dye from the extract to obtain colorless oat extract; taking colorless oat extract, and performing pressure filtration through a 300kDa nanofiltration tube to remove all components with the relative molecular mass lower than 300kDa in the extract, thereby obtaining high molecular weight oat beta-glucan extract; vacuum freeze drying the high molecular weight oat beta-glucan extracting solution to obtain high molecular weight oat beta-glucan extract powder; determining the relative molecular mass of beta-glucan in the obtained extract to be 349.7kDa by adopting GPC; the alpha-amylase and the beta-glucanase are adopted to verify that the high molecular weight oat beta-glucan in the extract can be degraded by the beta-glucanase but can not be degraded by the alpha-amylase; the content of high molecular weight beta-glucan in the obtained extract is 97.7 percent by adopting a dye Congo red colorimetric method.
Example 3
Selecting complete fresh oat, cleaning, airing, crushing, and sieving with a 60-mesh sieve to obtain oat powder; weighing oat powder, adding purified water according to a material-liquid ratio of 1 to 35, performing reflux extraction at 80 ℃ for 100min, filtering, extracting twice, combining filtrates, and standing; concentrating the filtrate under reduced pressure to 1/3 of the filtrate volume to obtain concentrated solution; placing the supernatant in a centrifuge, centrifuging at 2500 Xg for 10min, and collecting supernatant; slowly pressurizing the supernatant, injecting the supernatant into a column filled with supramolecular gel NBP5, performing column chromatography, and removing high-molecular organic dye in the extract to obtain colorless oat extract; taking colorless oat extract, and performing pressure filtration through a 300kDa nanofiltration tube to remove all components with the relative molecular mass lower than 300kDa in the extract, thereby obtaining high molecular weight oat beta-glucan extract; vacuum freeze drying the high molecular weight oat beta-glucan extracting solution to obtain high molecular weight oat beta-glucan extract powder; the relative molecular mass of beta-glucan in the obtained extract is measured by GPC and is 350.2kDa; the alpha-amylase and the beta-glucanase are adopted to verify that the high molecular weight oat beta-glucan in the extract can be degraded by the beta-glucanase but can not be degraded by the alpha-amylase; the content of high molecular weight beta-glucan in the obtained extract is determined to be 98.6 percent by adopting a dye Congo red colorimetric method.
The effect of the high molecular weight oat beta-glucan extract is evaluated by constructing a glycolipid metabolic abnormality model induced by a mouse food-borne late glycation end product, and the model is established by the following steps:
modeling experiment of food-derived advanced glycation end product-induced abnormal glycolipid metabolism: selecting 7-week-old Lepr db/db 28 male mice, 35-40 g weight, lepr in littermate db/m Male mice, 7 mice, served as controls (db/m group), weighed 25-30 g. After 1 week of acclimatization, 7 Lepr cells were randomly selected db/db The male mice are divided into normal (LA) group, food-borne advanced glycation end product induced abnormal glycolipid metabolism model (HA) group, high molecular weight oat beta-glucan extract and food-borne advanced glycation end product co-intake (HOS) group, high molecular weight oat beta-glucan extract and food-borne advanced glycation end product intake (HOA) group every other day, and the LA group and db/m group are given common feed every day, and the other groups are given common feed every other dayThe general feed and food-borne late-stage saccharified end-product feed were given alternately, and the HOS group and the HOA group were added with high molecular weight oat beta-glucan extracts to the hot processed feed and the general feed, respectively, which were rich in food-borne late-stage saccharified end-product, at 7% of the dry weight of the feeds. The weight, food intake and blood sugar level of mice are monitored regularly, after the mice are raised for 8 weeks, fasting blood sugar (FBG) and blood fat level of the mice are measured, if the weight of the mice in HA group is more than or equal to 40g, the fasting blood sugar (FBG) is more than or equal to 400mg/dL, the blood sugar after Oral Glucose Tolerance Test (OGTT) for 15min is more than or equal to 500mg/dL, and the blood sugar after 120min is more than or equal to 450mg/dL, the success of modeling of the glycolipid metabolic abnormality model induced by food-borne late-stage saccharification end products is determined.
Oral glucose tolerance (OGTT) test: for Lepr at week 8 of modeling experiment db/db Carrying out oral glucose tolerance (OGTT) test on male mice, after the mice are fasted and are not forbidden to be watered for 12 hours, intragastrically infusing glucose according to the amount of 1g/kg body weight, detecting blood glucose values respectively at 0min, 30min, 60min, 90min and 120min, making a blood glucose-time change curve graph, and calculating the area under the curve (AUC); at the end of an experiment, all mice are fasted for 12 hours without water prohibition, weighed, anesthetized by injecting 50mg/kg of pentobarbital sodium into the abdominal cavity, anticoagulated blood is collected from the heart, centrifugal separation is carried out for 10min at 3500 Xg, supernatant is taken and split charged, and the mixture is stored at-80 ℃ to be tested; and (3) detecting glycosylated hemoglobin (GHbA 1 c), fasting Blood Glucose (FBG) and Fasting Insulin (FINS) according to the kit specification, and weighing the weight of the liver, the kidney, the abdominal fat tissue, the perirenal fat tissue, the epididymal fat tissue and the mesenteric fat tissue after dissection.
The obtained oat beta-glucan extract with high molecular weight can remarkably inhibit the abnormal increase of body weight and blood sugar of the glycolipid metabolism abnormal model animal induced by the food-derived advanced glycation end product (figure 1 and figure 2A), remarkably relieve the impaired oral glucose tolerance (OGTT) (figure 2B-C), remarkably inhibit insulin resistance (figure 3A) and remarkably inhibit the fat accumulation in abdominal fat tissue, perirenal fat tissue, epididymal fat tissue and mesenteric fat tissue of the glycolipid metabolism abnormal model animal induced by the food-derived advanced glycation end product (figure 4) when being taken together with the food-derived advanced glycation end product and taken every other day with the food-derived advanced glycation end product. Has the effects of preventing and relieving abnormal glycolipid metabolism induced by food-derived advanced glycation end products, and can be used as food, feed or raw materials of human and animal medicines.
The results show that compared with the normal group (LA) mice, the body weight of the model group (HA) animals abnormally increases by 15%, the oral glucose tolerance is reduced by 22%, the contents of the glycosylated hemoglobin, the fasting blood glucose and the fasting insulin are respectively and obviously increased by 21%, 27% and 45%, and the weights of the abdominal cavity adipose tissues, the perirenal adipose tissues and the epididymal adipose tissues are respectively increased by 29%, 18% and 33%. The high molecular weight oat beta-glucan extract can be taken together with the food-derived advanced glycation end product (HOS group) and taken every other day with the food-derived advanced glycation end product (HOA group) to remarkably relieve the weight gain, wherein the average weight of mice in the HOS group and the HOA group is respectively reduced by 25 percent and 19 percent, the impaired degree of oral glucose tolerance can be relieved, and the increase of glycated hemoglobin, fasting blood glucose, fasting insulin, insulin resistance index and insulin sensitivity index can be remarkably inhibited; reduce the weight of abdominal adipose tissue, perirenal adipose tissue, epididymal adipose tissue and mesenteric adipose tissue.
The high molecular weight oat beta-glucan extract is used as a food, feed or pharmaceutical raw material for preventing and/or relieving glycolipid metabolism abnormality.
The amount of oat beta-glucan extract with high molecular weight to be used for preparing food is converted according to the following method: the required intake of oat beta-glucan extract per day was 7% of the dry weight of the feed, i.e. 490mg, calculated as 40g of the weight of one mouse, and the daily intake of mice was 12.25g/kg of body weight. The daily intake of human is 31.85mg/kg body weight according to the conversion coefficient of a body surface area method, and the dosage of the oat beta-glucan extract food is 0.3-5 g per day in terms of the weight of a child to an adult (10-150 kg).
Based on abnormal weight increase and impaired oral glucose tolerance of animals in the food-borne advanced glycation end product-induced glycolipid metabolic abnormality model group, the glycation hemoglobin, the fasting blood glucose, the fasting insulin, the insulin resistance index and the insulin sensitivity index are obviously increased; the weight of abdominal adipose tissue, perirenal adipose tissue, epididymal adipose tissue and mesenteric adipose tissue increased dramatically. The supplement mode of the high molecular weight oat beta-glucan extract which is taken together with the food-derived advanced glycation end product and taken every other day with the food-derived advanced glycation end product can obviously inhibit the abnormal increase of the body weight and the blood sugar of the glycolipid metabolic abnormality model animal induced by the food-derived advanced glycation end product, obviously relieve the impaired oral glucose tolerance, obviously inhibit the insulin resistance and obviously inhibit the fat in the abdominal fat tissue, the perirenal fat tissue, the epididymis fat tissue and the mesenteric fat tissue of the glycolipid metabolic abnormality model animal induced by the food-derived advanced glycation end product.
Claims (10)
1. The application of the high-molecular-weight oat beta-glucan extract is characterized in that the relative molecular mass of the oat beta-glucan extract is more than or equal to 300kDa, the purity of the oat beta-glucan extract is more than or equal to 95 percent, and the application refers to the application of the high-molecular-weight oat beta-glucan extract in relieving glycolipid metabolic abnormality induced by food-borne advanced glycation end products.
2. Use according to claim 1, characterized in that said high molecular weight oat β -glucan extract is prepared by:
(1) Extracting oat powder with water at 70-90 ℃, filtering, concentrating and centrifuging the extracting solution to obtain supernatant;
(2) Performing column chromatography on the supernatant obtained in the step (1) to obtain a colorless oat extract;
(3) And (3) taking the colorless oat extract obtained in the step (2), performing pressure filtration through a 300kDa nanofiltration tube, and drying to obtain the high-molecular-weight oat beta-glucan extract.
3. Use according to claim 2, wherein the oat flour in step (1) is obtained by: selecting oat, cleaning, drying in the air, crushing, and sieving with a 60-mesh sieve to obtain oat powder.
4. The use according to claim 2, wherein the mass ratio of the oat flour to water in step (1) is 1.
5. The use according to claim 2, wherein in step (1), the filtrate is concentrated to 1/3 to 1/4 of the original volume; centrifuging for 5-10 min at the rotating speed of 2500-3000 Xg.
6. The use according to claim 2, wherein the column chromatography in step (2) is a supramolecular gel column NBP5.
7. Use according to claim 1, wherein the high molecular weight oat β -glucan extract is used in the preparation of food, feed or medicament for preventing and/or alleviating food-borne advanced glycation end-product-induced dysglycolipid metabolism.
8. A food having an effect of preventing and/or alleviating glycolipid metabolic abnormalities induced by food-borne late glycation end products, characterized by comprising a high molecular weight oat β -glucan extract, the intake of which is 0.3-5 g/day.
9. A feed with the effect of preventing and/or relieving glycolipid metabolic abnormality induced by food-borne late-stage saccharification end products is characterized by comprising a high-molecular-weight oat beta-glucan extract, wherein the addition amount of the high-molecular-weight oat beta-glucan extract in the feed is 1% -10% of the dry weight of the feed.
10. A medicine with the effect of preventing and/or relieving glycolipid metabolic abnormality induced by food-borne late-stage saccharification end products is characterized by comprising a high-molecular-weight oat beta-glucan extract, wherein the content of the high-molecular-weight oat beta-glucan extract is 20% -50% of the content of medicine components.
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