CN115486505A - Marine algae polysaccharide health-care beverage with auxiliary blood sugar reducing effect and preparation method thereof - Google Patents

Marine algae polysaccharide health-care beverage with auxiliary blood sugar reducing effect and preparation method thereof Download PDF

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CN115486505A
CN115486505A CN202211148633.8A CN202211148633A CN115486505A CN 115486505 A CN115486505 A CN 115486505A CN 202211148633 A CN202211148633 A CN 202211148633A CN 115486505 A CN115486505 A CN 115486505A
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杜国丰
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Yingkou Institute of Technology
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Abstract

The invention discloses a marine algae polysaccharide health-care beverage with an auxiliary blood sugar reducing effect and a preparation method thereof. Compared with the prior art, the enteromorpha prolifera sulfated polysaccharide, the momordica charantia polysaccharide and the Chinese herbal medicines have the obvious effects of assisting in reducing blood sugar, tonifying qi and nourishing yin, promoting the production of body fluid to quench thirst, tonifying spleen, lung and kidney, resisting fatigue, removing vivotoxin and the like through the synergistic effect of the enteromorpha prolifera sulfated polysaccharide, the momordica charantia polysaccharide and the Chinese herbal medicines, are safe, have no side effect, are convenient to eat, and have better market development and application prospects.

Description

Marine algae polysaccharide health-care beverage with auxiliary blood sugar reducing effect and preparation method thereof
Technical Field
The invention relates to the technical field of health-care beverages, in particular to a marine algae polysaccharide health-care beverage with an auxiliary blood sugar reducing effect and a preparation method thereof.
Background
Enteromorpha is a large-scale natural wild green algae, grows on offshore shoals, and is widely distributed in rock ponds at estuaries and tidal zones. Enteromorpha is considered to be a dual-purpose algae from ancient times, and the Enteromorpha sulfated polysaccharide is a linear polymer extracted from the Enteromorpha, and the main stem of the Enteromorpha sulfated polysaccharide is a D-GlcAp-alpha- (1 → 4) -3-sulfate-L-Rhap-beta- (1 → 4) -D-Xylp-beta- (1 → 4) -3-sulfate-L-Rhap unit. The enteromorpha sulfuric acid polysaccharide has various biological activities, including blood sugar reduction, tumor resistance, gastrointestinal tract protection, anticoagulant property, oxidation resistance and immunoregulation. Therefore, the enteromorpha sulfuric acid polysaccharide has excellent development prospect in the field of health-care food.
The water-soluble momordica polysaccharide is an acidic polysaccharide extracted from momordica charantia pulp, mainly comprises arabinose, galacturonic acid, rhamnose and xylose, and related researches show that the momordica charantia polysaccharide with antioxidant activity can reduce the blood sugar level of diabetic mice, so that the momordica charantia polysaccharide is widely applied to the field of medicines and modern food industry.
Type 2 diabetes (T2 DM), also known as non-insulin dependent diabetes mellitus or adult onset diabetes, is characterized by abnormal insulin secretion, overproduction of glucose by the liver, and insulin resistance in peripheral tissues such as the liver and skeletal muscle. Chronic hyperglycemia in diabetes is associated with long-term damage, dysfunction and failure of various organs, especially the eyes, kidneys, nerves, heart and blood vessels. The risk factors for type 2 diabetes are relatively well known and include age, obesity, lifestyle, dietary patterns and genetic environmental interactions. Most hypoglycemic agents used to treat T2DM have an insulin-dependent mechanism of action, i.e., stimulation of insulin production (e.g., sulfonylureas, glinides, incretins, and dipeptidyl peptidase-4 inhibitors), enhancement of insulin sensitivity (e.g., thiazolidinediones and biguanides), or direct enhancement of endogenous insulin levels (e.g., basal insulin and prandial insulin). As the etiology and pathogenesis of diabetes are not fully elucidated, no medicine can cure diabetes at present, and patients need to take medicine for life to maintain blood sugar balance. In recent years, with the improvement of living standard and health care consciousness, natural bioactive products which are green, environment-friendly and widely available are applied to the development of health care food, and the health care food with the function of assisting in reducing blood sugar is very beneficial to the improvement of the living quality of diabetics and is favored by more and more patients.
Fasting plasma glucose (FBS) refers to the blood glucose value detected by blood plasma collected before breakfast after fasting (at least 8-10 hours without any food and drinking water) at night, can reflect the function of islet beta cells, generally represents the secretion function of basal insulin, and is the most common detection index for diabetes. Clinically, blood glucose refers to glucose in the blood. Each individual may have fluctuations in blood glucose levels throughout the day with food intake, activity, etc. Blood glucose levels generally at fasting stateCompared withIs constant. The method of glucose oxidase is adopted in clinical detection, so that the real blood glucose concentration can be specifically detected. Blood glucose concentration remains relatively stable as regulated by the nervous system and hormones. When these adjustments lose their original relative balance, hyperglycemia or hypoglycemia occurs. Therefore, measurement of the fasting blood glucose level can reflect the blood glucose metabolism of the body.
Glucose tolerance refers to the ability of the body to regulate blood glucose concentration. After eating rice, flour staple food or high-sugar food, the normal people are almost completely absorbed by intestinal tracts, so that the blood sugar is increased, the insulin secretion is stimulated, the synthesis of liver glycogen is increased, the decomposition is inhibited, the output of liver glycogen is reduced, and the utilization of glucose by tissues in vivo is increased, therefore, the highest blood sugar after meals does not exceed 10.0mmol/L, and more or less blood sugar is kept in a relatively stable range after eating. This indicates that normal persons have strong glucose tolerance, i.e., normal glucose tolerance. However, in the case of a person with insufficient insulin secretion, the concentration of insulin may exceed 7.8mmol/L and may be equal to or greater than 11.1 mmol/L2 hours after 75g of glucose is orally administered, indicating that the person has impaired glucose tolerance.
The immunohistochemical staining is to utilize the characteristic of combining known antibody and cell antigen specificity, to make the display agent marked on the antibody display certain color through chemical reaction, and to observe with the aid of microscope, fluorescence microscope or electron microscope, so as to achieve the purpose of quantitative and positioning analysis of chemical components in tissue and cell structure.
The insulin receptor, the instr gene, insulin, functions by interacting with the instr on the cell membrane (mainly in hepatocytes, adipocytes and muscle cells) depending on the receptor density and affinity on the cell membrane. Therefore, insR is the first key point of insulin action, and InsR genes are crucial for the study of insulin resistance.
Glucokinase (GCK), also known as hexokinase, is a key enzyme in the regulation of glucose metabolism and insulin secretion, and is the first rate-limiting enzyme in the glycolysis process. It catalyzes the phosphorylation of glucose to glucose-6-phosphate, a process that is a prerequisite for the synthesis of hepatic glycogen. Loss of function or reduced expression of GCK inhibits glucose utilization by the liver and reduces insulin secretion by the pancreatic islet beta cells. Under conditions of high fat diet-induced insulin resistance, GCK is critical for beta cell regeneration. Mutations in the GCK gene may be a cause of diabetes. Therefore, the increase of the expression level of GCK gene leads to a certain control of blood glucose concentration.
China is the origin of Chinese herbal medicine, and Chinese herbal medicine resources occupy monopoly advantages. The intensive exploration, research and summary of Chinese herbal medicines and traditional Chinese medicines by ancient Xianxian make the Chinese herbal medicines be most widely recognized and applied. The chemical components contained in the Chinese herbal medicine are complex, and generally comprise saccharides, amino acids, proteins, oil, wax, enzyme, pigment, vitamins, organic acids, tannin, inorganic salt, volatile oil, alkaloid, glycosides and the like. Each Chinese medicinal herb may contain multiple components. Some of these components have obvious biological activity and are used for medical treatment, and are often called as effective components, such as alkaloids, glycosides, volatile oils, amino acids, etc. The analysis and detection research of modern traditional Chinese medicine and pharmacy discovers that functional food developed by utilizing Chinese herbal medicines contains a large amount of bioactive components, and the functional food can regulate the metabolic function of a human body, improve the immunoregulation capability, remove free radicals, delay senility and the like after being reasonably eaten for a long time. In recent years, chinese herbal medicine compound preparations are increasingly used for preventing and assisting in reducing blood sugar and preventing diabetic complications, but the application effect of the Chinese herbal medicine compound preparation needs to be further improved urgently.
At present, various functional health foods added with Chinese herbal medicine components have the problems of insignificant efficacy, uncertain efficacy components and the like in the eating process, and the maximum level of the efficacy of the traditional Chinese herbal medicine is not exerted. Aiming at the problems, the invention develops a marine algae polysaccharide health-care beverage which utilizes green algae enteromorpha sulfuric acid polysaccharide, bitter gourd polysaccharide and compound plant Chinese herbal medicines and has the auxiliary blood sugar reducing effect, and meets the market demands on marine algae functional foods and Chinese herbal medicine health-care foods.
Disclosure of Invention
The invention aims to solve the problem that the effect of the existing functional food is not obvious, and provides a marine algae polysaccharide health-care beverage with the effect of assisting in reducing blood sugar and a preparation method thereof.
In order to achieve the purpose, the invention is implemented according to the following technical scheme:
the invention provides a preparation method of a marine algae polysaccharide health-care beverage with an auxiliary blood sugar reducing effect, which comprises the following steps:
s1, chinese herbal medicine pretreatment: the following Chinese herbal medicines are taken according to parts by weight: 15-20 parts of wolfberry fruit, 10-15 parts of Tiegun yam, 10-20 parts of rhizoma polygonati, 10-15 parts of corn stigma, 5-8 parts of astragalus membranaceus, 3-5 parts of epimedium, 3-5 parts of radix puerariae, 4-6 parts of radix ophiopogonis, 2-3 parts of radix polygonati officinalis and 3-5 parts of rhizoma anemarrhenae, and the raw materials are respectively cleaned by purified water and dried in the air;
s2, crushing: crushing Dioscorea opposita Thunb and fructus Lycii with a crusher, pulverizing the rest Chinese medicinal materials, and sieving with 200 mesh sieve;
s3, chinese herbal medicine leaching: after the Chinese herbal medicines in the step S2 are uniformly mixed, purified water with the mass of 150 times that of the Chinese herbal medicines is added, and the mixture is heated and decocted for 40-60min; filtering to obtain Chinese medicinal extractive solution;
s4, compounding: adding 10-20 parts of green alga enteromorpha sulfuric acid polysaccharide and 10-20 parts of balsam pear polysaccharide into the Chinese herbal medicine leaching liquor, and stirring until the green alga enteromorpha sulfuric acid polysaccharide and the balsam pear polysaccharide are completely dissolved;
s5, centrifuging: centrifuging to remove precipitate to obtain mixed medicinal liquid;
s6, blending: adding xylo-oligosaccharide 1 wt%, citric acid 0.5 wt%, xylitol 0.5 wt%, calcium lactate 1 wt%, and Vc 0.2 wt% into the mixed medicinal liquid, and adjusting pH to 7.0-7.5; homogenizing in a homogenizer;
s7, fine filtration: filtering the homogenized health beverage with 200 mesh duplex filter, and fine filtering with diatom filter to remove impurities and insoluble substances;
s8, sterilization: performing high-temperature instantaneous sterilization on the prepared health-care beverage;
s9, filling: vacuum packaging, and cooling to normal temperature to obtain the final product.
Preferably, in the step S1, the following Chinese herbal medicines are taken according to parts by weight: 15 parts of green alga enteromorpha sulphated polysaccharide, 15 parts of balsam pear polysaccharide, 15 parts of medlar, 12 parts of Tiegun yam, 15 parts of rhizoma polygonati, 12 parts of corn stigma, 6 parts of astragalus membranaceus, 4 parts of epimedium, 4 parts of radix puerariae, 5 parts of radix ophiopogonis, 2.5 parts of polygonatum odoratum and 4 parts of rhizoma anemarrhenae.
Preferably, the centrifugation in step S5 is carried out at 5000r/min for 10-15min.
Preferably, the homogenization pressure of the step S6 is 25MPa, and the temperature is 60-70 ℃.
Preferably, the sterilization temperature of the step S8 is 120-140 ℃ and the time is 4-6S.
Preferably, the vacuum degree of the vacuum filling in the step S9 is 0.04-0.05Mpa, and the filling temperature is 60-75 ℃.
The second purpose of the invention is to provide a marine algae polysaccharide health-care beverage with the auxiliary blood sugar reducing effect, which is prepared by the preparation method.
Compared with the prior art, the enteromorpha prolifera polysaccharide sulfate, the polysaccharide balsam pear and the Chinese herbal medicines have the obvious effects of assisting in reducing blood sugar, tonifying qi and nourishing yin, promoting the production of body fluid to quench thirst, tonifying spleen, lung and kidney, resisting fatigue, removing vivotoxin and the like through the synergistic effect of the enteromorpha prolifera polysaccharide sulfate, the polysaccharide balsam pear and the Chinese herbal medicines, are safe, have no side effect, are convenient to eat, and have better market development and application prospects.
Drawings
FIG. 1 is a graph of the effect of each treatment group on fasting blood glucose levels in diabetic model mice.
Fig. 2 is a graph of the effect of each treatment group on oral glucose tolerance in diabetic model mice.
FIG. 3 shows the islet tissue morphology of mice in each test group observed with hematoxylin-eosin (H & E) staining light microscope: (a) a normal control group; (b) streptozotocin model group; (c) metformin hydrochloride group; (d) momordica polysaccharide and Chinese herbal medicine; (e) algal polysaccharides group; (f) algal polysaccharide, momordica polysaccharide and a Chinese herbal medicine.
FIG. 4 is a graph showing the effect of each treatment group on the mRNA expression levels of the GCK and InsR genes in the liver of diabetic mice.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. The specific embodiments described herein are merely illustrative of the invention and do not delimit the invention.
The Chinese herbal medicines and test reagents used in the following examples were purchased from the market except for the specific instructions.
Example 1
S1, chinese herbal medicine pretreatment: weighing the following Chinese herbal medicines: 15 g of medlar, 10 g of Tiegun yam, 10 g of rhizoma polygonati, 10 g of corn stigma, 5g of astragalus, 3 g of epimedium, 3 g of kudzu root, 4 g of radix ophiopogonis, 2g of polygonatum and 3 g of rhizoma anemarrhenae, which are respectively washed by purified water and dried in the air;
s2, crushing: crushing Dioscorea opposita Thunb and fructus Lycii with a crusher, pulverizing the rest Chinese medicinal materials, and sieving with 200 mesh sieve;
s3, chinese herbal medicine leaching: after the Chinese herbal medicines in the step S2 are uniformly mixed, 9750 g of purified water is added, and the mixture is heated and decocted for 40min; filtering to obtain Chinese medicinal leaching solution;
s4, compounding: adding 10 g of green alga enteromorpha sulfated polysaccharide and 10 g of balsam pear polysaccharide into the Chinese herbal medicine leaching liquor, and stirring until the green alga enteromorpha sulfated polysaccharide and the balsam pear polysaccharide are completely dissolved;
s5, centrifugation: centrifuging in a centrifuge of 5000r/min for 10min to remove precipitate to obtain mixed medicinal liquid;
s6, blending: adding 98 g of xylo-oligosaccharide, 49 g of citric acid, 49 g of xylitol, 98 g of calcium lactate and 19.6 g of Vc into the mixed liquid medicine, and adjusting the pH value to 7.0 (the pH value is adjusted by using citric acid and sodium citrate which are commonly used in the field of beverages); homogenizing in a homogenizer with pressure of 25MPa and temperature of 60 ℃;
s7, fine filtration: filtering the homogenized health beverage with 200 mesh duplex filter, and fine filtering with diatom filter to remove impurities and insoluble substances;
s8, sterilization: instantly sterilizing the prepared health beverage at 120 deg.C for 6s;
s9, filling: vacuum packaging at vacuum degree of 0.04Mpa and packaging temperature of 75 deg.C, and cooling to normal temperature to obtain marine algae polysaccharide health beverage with auxiliary blood sugar lowering effect.
Example 2
S1, chinese herbal medicine pretreatment: weighing the following Chinese herbal medicines: 20 g of medlar, 15 g of dioscorea opposita thunb, 20 g of rhizoma polygonati, 15 g of corn stigma, 8 g of astragalus membranaceus, 5g of epimedium, 5g of radix puerariae, 6 g of radix ophiopogonis, 3 g of radix polygonati officinalis and 5g of rhizoma anemarrhenae, and the raw materials are respectively washed by purified water and dried in the air;
s2, crushing: crushing rhizoma Dioscoreae and fructus Lycii with a pounder, pulverizing the rest Chinese medicinal materials, and sieving with 200 mesh sieve;
s3, chinese herbal medicine leaching: after the Chinese herbal medicines in the step S2 are uniformly mixed, 15300 g of purified water is added, and the mixture is heated and decocted for 60min; filtering to obtain Chinese medicinal extractive solution;
s4, compounding: adding 20 g of green alga enteromorpha sulfated polysaccharide and 20 g of balsam pear polysaccharide into the Chinese herbal medicine leaching liquor, and stirring until the green alga enteromorpha sulfated polysaccharide and the balsam pear polysaccharide are completely dissolved;
s5, centrifugation: centrifuging in a centrifugal machine of 5000r/min for 15min to remove precipitate to obtain mixed medicinal liquid;
s6, blending: adding 154 g of xylo-oligosaccharide, 77 g of citric acid, 77 g of xylitol, 154 g of calcium lactate and 30.8 g of Vc into the mixed liquid medicine, and adjusting the pH value to 7.5 (the pH value is adjusted by selecting citric acid and sodium citrate commonly used in the field of beverages); homogenizing in a homogenizer with pressure of 25MPa and temperature of 70 deg.C;
s7, fine filtration: filtering the homogenized health beverage with 200 mesh duplex filter, and fine filtering with diatom filter to remove impurities and insoluble substances;
s8, sterilization: instantly sterilizing the blended health beverage at 140 deg.C for 4s;
s9, filling: vacuum filling is carried out, the vacuum degree is 0.05Mpa, the filling temperature is 60 ℃, and the marine algae polysaccharide health drink with the auxiliary blood sugar reducing effect is obtained after cooling to the normal temperature.
Example 3
S1, chinese herbal medicine pretreatment: weighing the following Chinese herbal medicines: 15 g of medlar, 12 g of dioscorea opposita thunb, 15 g of rhizoma polygonati, 12 g of corn stigma, 6 g of astragalus membranaceus, 4 g of epimedium, 4 g of radix puerariae, 5g of radix ophiopogonis, 2.5 g of polygonatum odoratum and 4 g of rhizoma anemarrhenae, and the medlar, the rhizoma dioscoreae nipponicae thunb and the rhizoma polygonati are respectively washed by purified water and dried;
s2, crushing: crushing rhizoma Dioscoreae and fructus Lycii with a pounder, pulverizing the rest Chinese medicinal materials, and sieving with 200 mesh sieve;
s3, chinese herbal medicine extraction: after the Chinese herbal medicines in the step S2 are uniformly mixed, purified water with the mass of 150 times that of the Chinese herbal medicines is added, and the mixture is heated and decocted for 50min; filtering to obtain Chinese medicinal extractive solution;
s4, compounding: adding 15 g of green alga enteromorpha sulfated polysaccharide and 15 g of balsam pear polysaccharide into the Chinese herbal medicine leaching liquor, and stirring until the green alga enteromorpha sulfated polysaccharide and the balsam pear polysaccharide are completely dissolved;
s5, centrifugation: centrifuging in a centrifuge of 5000r/min for 12min to remove precipitate to obtain mixed medicinal liquid;
s6, blending: adding 119 g of xylooligosaccharide, 59.5 g of citric acid, 59.5 g of xylitol, 119 g of calcium lactate and 23.8 g of Vc into the mixed liquid medicine, and adjusting the pH value to 7.2 (the pH value is adjusted by using citric acid and sodium citrate which are commonly used in the field of beverages); homogenizing in a homogenizer with pressure of 25MPa and temperature of 65 deg.C;
s7, fine filtration: filtering the homogenized health beverage with 200 mesh duplex filter, and fine filtering with diatom filter to remove impurities and insoluble substances;
s8, sterilization: instantly sterilizing the blended health beverage at 130 deg.C for 5s;
s9, filling: vacuum filling at 70 deg.C under 0.045Mpa, and cooling to normal temperature to obtain marine algae polysaccharide health beverage with auxiliary blood sugar lowering effect.
Further, in order to verify that the health drink prepared by the present invention has the auxiliary hypoglycemic effect, the hypoglycemic effect evaluation of the marine algae polysaccharide health drink (the health drink prepared in example 3) was performed by the mouse animal test as follows.
Male Kunming mice (22 + -2 g in body weight) were randomly divided into 10 normal control groups and 60 model groups. The control group is fed with normal feed, the model group is fed with high-sugar and high-fat feed for 4 weeks continuously, after 4 weeks, the model group is injected with streptozotocin according to the dosage of 30mg/kg, the fasting blood glucose level of the mouse is measured after 5 days, the blood glucose value reaches more than 11.1mmol/L, and the model building success of the hyperglycemic mouse model is proved. The hyperglycemic mice were divided into 5 groups (streptozotocin model group, metformin hydrochloride group, charantin + Chinese herbal medicine, algal polysaccharide group, algal polysaccharide + charantin + Chinese herbal medicine), 10 mice each. The metformin hydrochloride group is intragastrically filled with 500mg/kg metformin hydrochloride, the momordica polysaccharide + Chinese herbal medicine, the algal polysaccharide group, the algal polysaccharide + momordica polysaccharide + Chinese herbal medicine are intragastrically filled with 500mg/kg momordica polysaccharide + Chinese herbal medicine, the algal polysaccharide + momordica polysaccharide + Chinese herbal medicine aqueous solution respectively, the intragastrically filled distilled water is filled for 1 time every day, and the equivalent amount of intragastrically filled distilled water is filled in a normal control group and a streptozotocin model group. The intervention cycle was 4 weeks, with the following tests during and after the intervention: (1) On the day of intervention, and on the 14 th and 28 th days after the intervention (after the intervention is finished), blood is collected from the tail vein of the mouse after fasting for 12 hours, and fasting blood glucose is measured by a glucometer; (2) After the intervention is finished, each group of mice is perfused with 50% glucose solution according to 2g/kg, and the blood sugar values after 0.5h, 1h and 2h of perfusion before the perfusion are measured; (3) Taking mouse pancreatic gland tissues, placing the mouse pancreatic gland tissues in 8% paraformaldehyde solution for fixing for 24 hours, embedding the mouse pancreatic gland tissues in conventional paraffin, continuously slicing the mouse pancreatic gland tissues to the thickness of 6 microns, taking 1 wax tape every 20 pieces, mounting the wax tape on a glass slide coated with APES (advanced plasma enhanced imaging System), performing immunohistochemical staining by utilizing hematoxylin-eosin (H & E), and observing the shapes of islet cells by using a light microscope; (4) Taking liver tissues of each group of mice, homogenizing under an ice bath environment, extracting total RNA by using a kit of TaKaRa company, and detecting the mRNA levels of GCK and InsR in the liver tissues of each group of mice by combining a reverse transcriptase kit with a real-time fluorescent quantitative PCR technology.
The results of the fasting blood glucose levels (FBS) in mice by the gavage test are shown in FIG. 1. As can be seen from the figure, before the gavage intervention (day 0), the fasting blood glucose of the streptozotocin model group, the metformin hydrochloride group, the momordica polysaccharide + Chinese herbal medicine group, the algal polysaccharide + momordica polysaccharide + Chinese herbal medicine group is obviously higher than that of the normal control group (P < 0.05). After 2 weeks (14 days) of gavage intervention, FBS levels declined in each intervention group with a descending trend in order: the metformin hydrochloride group is greater than algal polysaccharide, momordica charantia polysaccharide and a Chinese herbal medicine group, the algal polysaccharide group is greater than momordica charantia polysaccharide and the Chinese herbal medicine group. After 4 weeks (28 days) of the gavage intervention, the FBS levels of the metformin hydrochloride group, the momordica polysaccharide + Chinese herbal medicine group, the algal polysaccharide group and the algal polysaccharide + momordica polysaccharide + Chinese herbal medicine group were significantly decreased (P < 0.05) compared to those before the gavage intervention (day 0), and the FBS levels of the algal polysaccharide + momordica polysaccharide + Chinese herbal medicine group were equivalent to those of the metformin hydrochloride group (FBS were 6.77mmol/L and 6.25mmol/L, respectively).
The results of the Oral Glucose Tolerance Test (OGTT) by gavage test are shown in fig. 2. As can be seen from the figure, the blood glucose of each group of mice reached a peak value within 1 hour after glucose injection, and blood glucose was substantially recovered within 2 hours. Compared with the streptozotocin model group, the metformin hydrochloride group, the momordica polysaccharide and the Chinese herbal medicine, the algal polysaccharide group and the algal polysaccharide and the momordica polysaccharide and the Chinese herbal medicine have quicker recovery of blood sugar, but the glucose tolerance of the algal polysaccharide, the momordica polysaccharide and the Chinese herbal medicine is almost the same as that of the metformin hydrochloride group, and the recovery of the blood sugar is faster (P is less than 0.05) compared with the momordica polysaccharide and the Chinese herbal medicine.
The results of immunohistochemical staining of pancreatic tissues of mice of each experimental group by hematoxylin-eosin (H & E) and observation of islet cell morphology by light microscopy are shown in fig. 3. Pancreatic acinar and islet cells of the control group were intact (fig. 3 a); the islets of mice in the streptozotocin model group are distributed dispersedly, the volume is obviously reduced, most of the islet cells are atrophied, the staining is light, the number of the cells is small, the cytoplasm is obviously reduced, the cell nucleus is greatly condensed, and the lymphocytes widely permeate and invade the islets (figure 3 b); compared with the streptozotocin model group, the positive control metformin hydrochloride group has the advantages that although the shape of the pancreatic islets is still irregular, the number of internal cells is obviously increased; the nuclear condensation phenomenon was not well restored, the nuclear staining was deeper, but the cytoplasmic distribution was still uneven (fig. 3 c); the shapes of the islets of the diabetes model mice after the gavage dry prognosis of the momordica polysaccharide, the Chinese herbal medicine, the algal polysaccharide and the algal polysaccharide, the momordica polysaccharide and the Chinese herbal medicine are all changed to be normal, but the differences exist among the groups (figures 3d-3 f); the algal polysaccharide group (fig. 3 e) has more regular islet edges, the necrotic cells in the islets are recovered, the number of the cells is obviously increased, but the cells are arranged irregularly, the phenomenon of nuclear condensation is relieved, the cytoplasm is distributed more uniformly, and the staining is lighter; the islet cells of algal polysaccharide + momordica polysaccharide + Chinese herbal medicine were significantly increased, with essentially normal morphology and structure, homogeneous cytoplasm, high number, clear nucleoli, and occasionally small amount of lymphocyte infiltration (fig. 3 f).
The mRNA expression level of GCK and InsR in the liver tissue of each group of mice is detected by a molecular biology real-time fluorescent quantitative PCR technology. The results are shown in fig. 4, after 28 days of gastric lavage treatment, compared with the streptozotocin model group, mRNA expression levels of the intervention groups instr are increased averagely, wherein the increase levels of the positive control metformin hydrochloride group and the combination of algal polysaccharide + momordica polysaccharide + Chinese herbal medicine are more significant (P is less than 0.05). Meanwhile, the mRNA expression level of GCK of each intervention group is greatly increased (P < 0.05), wherein the increase level of a positive control metformin hydrochloride group and the seaweed polysaccharide, the momordica polysaccharide and the Chinese herbal medicine group reaches a very significant level (P < 0.01), the difference between the two groups has no statistical significance, but the difference between the seaweed polysaccharide, the momordica polysaccharide and the Chinese herbal medicine group, the momordica polysaccharide, the Chinese herbal medicine group and the seaweed polysaccharide group is significant (P < 0.05).
In conclusion, the invention has the advantages that the green alga enteromorpha sulphated polysaccharide, the balsam pear polysaccharide and a plurality of Chinese herbal medicines are scientifically combined, the synergistic effect among the algal polysaccharide, the balsam pear polysaccharide and the Chinese herbal medicines can effectively assist the effects of reducing blood sugar, tonifying qi and nourishing yin, promoting the production of body fluid to quench thirst, tonifying spleen, lung and kidney, resisting fatigue, removing vivotoxin and the like, and the green alga enteromorpha sulphated polysaccharide-balsam pear polysaccharide-Chinese herbal medicine is safe, free of side effect and convenient to eat. Different Chinese herbal medicines have different properties and tastes, the effects of the Chinese herbal medicines are greatly different, the Chinese herbal medicines have complex interaction relationship after being combined, the Chinese herbal medicines are closely matched, and the monarch, minister, assistant and guide relationships are substantially changed by different matching ratios, so that the relationship of the composite Chinese herbal medicine is substantially changed. According to the application, a complex interaction relationship exists among green alga enteromorpha sulphated polysaccharide, momordica charantia polysaccharide, chinese wolfberry, dioscorea opposita, rhizoma polygonati, corn stigma, astragalus membranaceus, epimedium herb, radix puerariae, radix ophiopogonis, radix polygonati officinalis, rhizoma anemarrhenae and other Chinese herbal medicines, and an unexpected technical effect is obtained through reasonable proportioning, so that the relationship of the marine alga polysaccharide health drink is substantially changed, and compared with blood sugar values (6.77 mmol/L) of a streptozotocin model group, a momordica charantia polysaccharide and a Chinese herbal medicine group and blood sugar values (23.14 mmol/L, 10.25mmol/L and 8.75 mmol/L) of a streptozotocin model group, a momordica charantia polysaccharide and a Chinese herbal medicine group and a seaweed polysaccharide group after the marine alga polysaccharide health drink is fed for 4 weeks, the blood sugar level is respectively reduced by 70.74%, 33.95% and 22.63%, and the product has better auxiliary blood sugar reduction activity; and the test result shows that the change of the raw material composition or the proportion of the marine algae polysaccharide health-care beverage can reduce the blood sugar reducing capability (P < 0.01) of the marine algae polysaccharide health-care beverage, and the synergistic effect among the green algae enteromorpha sulfated polysaccharide, the balsam pear polysaccharide and the Chinese herbal medicine plays a role in obviously improving the functional activity of the marine algae polysaccharide health-care beverage. As can be known from a 28-day gastric lavage test of a mouse, the physical sign and behavior of the mouse are normal, the body state is healthy, and the biochemical indexes of blood and blood do not change abnormally, so that the marine algae polysaccharide health drink is determined to belong to nontoxic foods, and people can safely and reassurantly eat the marine algae polysaccharide health drink.
The technical solution of the present invention is not limited to the limitations of the above specific embodiments, and all technical modifications made according to the technical solution of the present invention fall within the protection scope of the present invention.

Claims (7)

1. A preparation method of a marine algae polysaccharide health-care beverage with an auxiliary blood sugar reducing effect is characterized by comprising the following steps:
s1, chinese herbal medicine pretreatment: the following Chinese herbal medicines are taken according to parts by weight: 15-20 parts of wolfberry fruit, 10-15 parts of dioscorea opposita thunb, 10-20 parts of rhizoma polygonati, 10-15 parts of corn stigma, 5-8 parts of astragalus membranaceus, 3-5 parts of epimedium herb, 3-5 parts of radix puerariae, 4-6 parts of radix ophiopogonis, 2-3 parts of polygonatum odoratum and 3-5 parts of rhizoma anemarrhenae, and the raw materials are respectively cleaned by purified water and dried in the air;
s2, crushing: crushing rhizoma Dioscoreae and fructus Lycii with a pounder, pulverizing the rest Chinese medicinal materials, and sieving with 200 mesh sieve;
s3, chinese herbal medicine leaching: after the Chinese herbal medicines in the step S2 are uniformly mixed, purified water with the mass 150 times that of the Chinese herbal medicines is added, and the mixture is heated and decocted for 40-60min; filtering to obtain Chinese medicinal leaching solution;
s4, compounding: adding 10-20 parts of green alga enteromorpha sulfuric acid polysaccharide and 10-20 parts of balsam pear polysaccharide into the Chinese herbal medicine leaching liquor, and stirring until the green alga enteromorpha sulfuric acid polysaccharide and the balsam pear polysaccharide are completely dissolved;
s5, centrifugation: centrifuging to remove precipitate to obtain mixed medicinal liquid;
s6, blending: adding xylo-oligosaccharide 1 wt%, citric acid 0.5 wt%, xylitol 0.5 wt%, calcium lactate 1 wt%, and Vc 0.2 wt% into the mixed medicinal liquid, and adjusting pH to 7.0-7.5; homogenizing in a homogenizer;
s7, fine filtration: filtering the homogenized health beverage with 200 mesh duplex filter, and fine filtering with diatom filter to remove impurities and insoluble substances;
s8, sterilization: performing high-temperature instantaneous sterilization on the prepared health-care beverage;
s9, filling: vacuum filling is adopted, and the marine algae polysaccharide health-care beverage with the auxiliary blood sugar reducing effect is obtained after cooling to normal temperature.
2. The preparation method of the marine algae polysaccharide health drink with the auxiliary hypoglycemic effect according to claim 1, wherein in the step S1, the following Chinese herbal medicines are taken according to parts by weight: 15 parts of green alga enteromorpha sulphated polysaccharide, 15 parts of balsam pear polysaccharide, 15 parts of medlar, 12 parts of iron stick yam, 15 parts of rhizoma polygonati, 12 parts of corn stigma, 6 parts of astragalus membranaceus, 4 parts of epimedium, 4 parts of radix puerariae, 5 parts of radix ophiopogonis, 2.5 parts of polygonatum odoratum and 4 parts of rhizoma anemarrhenae.
3. The preparation method of the marine algae polysaccharide health beverage with the auxiliary hypoglycemic effect according to claim 1, wherein the preparation method comprises the following steps: and in the step S5, centrifuging at 5000r/min for 10-15min.
4. The preparation method of the marine algae polysaccharide health drink with the auxiliary hypoglycemic effect according to claim 1, wherein the preparation method comprises the following steps: the homogenizing pressure of the step S6 is 25MPa, and the temperature is 60-70 ℃.
5. The preparation method of the marine algae polysaccharide health beverage with the auxiliary hypoglycemic effect according to claim 1, wherein the preparation method comprises the following steps: the sterilization temperature of the step S8 is 120-140 ℃, and the time is 4-6S.
6. The preparation method of the marine algae polysaccharide health beverage with the auxiliary hypoglycemic effect according to claim 1, wherein the preparation method comprises the following steps: the vacuum degree of the vacuum filling in the step S9 is 0.04-0.05Mpa, and the filling temperature is 60-75 ℃.
7. A marine algae polysaccharide health beverage with auxiliary blood sugar lowering effect prepared by the preparation method of any one of claims 1-6.
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