CN106858603B

CN106858603B - New function of chitobiose and application of chitobiose in health-care food

Info

Publication number: CN106858603B
Application number: CN201510923633.4A
Authority: CN
Inventors: 江正强; 吴夏; 闫巧娟; 石玉琴; 王静
Original assignee: China Agricultural University
Current assignee: China Agricultural University
Priority date: 2015-12-14
Filing date: 2015-12-14
Publication date: 2021-04-06
Anticipated expiration: 2035-12-14
Also published as: CN106858603A

Abstract

The present invention relates to a new function of chitobiose and its application in health-care food. The chitobiose is a degradation product of chitin, and is an oligosaccharide formed by connecting 2N-acetylglucosamines by glycosidic bonds. Animal experiments show that the chitobiose has obvious alpha-glucosidase inhibition activity, the IC50 of the chitobiose is 2.0mg/mL, the fasting blood sugar level of type II diabetic mice can be obviously reduced to 6.1mg/mL, the fasting blood sugar value of the chitobiose is obviously lower than that of a control acarbose group by 7.9mg/mL, and the fasting blood sugar value range of the control acarbose group is close to the upper limit of the fasting blood sugar value range of normal healthy mice. Second, chitobiose significantly reduces serum lipid levels and inhibits pancreatic lipid peroxidation levels compared to acarbose. In addition, chitobiose stabilizes the body weight of diabetic mice. The compounding of the chitobiose and the xylo-oligosaccharide also has obvious effect of reducing blood sugar. The chitobiose has good hypoglycemic effect, and can be used for developing safe and effective health-care food for preventing and improving type II diabetes.

Description

New function of chitobiose and application of chitobiose in health-care food

Technical Field

The invention relates to the field of health-care food, in particular to a novel function of chitobiose and application thereof in health-care food. Especially to the function of chitobiose in preventing or improving type II diabetes and the application in health food.

Background

Chitin (Chitin, also known as Chitin, chitosan or Chitin) is a water-insoluble linear polymer formed by connecting N-acetylglucosamine by beta-1, 4-glycosidic bonds, is a renewable biomass with the content inferior to that of cellulose in nature, and is synthesized in the world at up to billions of tons per year.

Chitin is widely found in the carapace of shrimps, crabs and insects, as well as in the cell walls of fungi and plants and some green algae. Chitin is degraded to produce chitooligosaccharides, such as chitooligosaccharides (with a degree of polymerization of 2-10 acetyl-containing monosaccharides) produced by hydrolysis of the beta-1, 4-glycosidic linkages in the chitin chain (Bhattacharya, D., Nagpure, A.and Gupta, R.K., Bacterial chitinases: properties and potential, clinical Reviews in Biotechnology,2007,27(1): 21-28). Common chitooligosaccharides include chitobiose (N-acetylchitobiose), chitotriose (N-acetylchitotriose), and the like.

The preparation method of the chitosan oligosaccharide generally adopts a chemical method (concentrated hydrochloric acid hydrolysis) or an enzymatic method, and compared with the preparation method of the chitosan oligosaccharide by adopting an enzyme (biological enzyme) method, the preparation method is safer; the chitosan oligosaccharide is formed by hydrolyzing chitosan obtained by deacetylation treatment of chitin.

In recent decades, the research on chitin and chitosan has been widely carried out at home and abroad, and the physiological functions of chitin and chitosan have also been reported, for example, the chitin and chitosan are used as raw materials to prepare band-aid, biological membranes and medical sutures for medical treatment, or have the effects of resisting tumors and preventing and treating pathogenic biological infection (Jixiang, Yao soldier, Zhuangyi Qing, etc.. the biological sterilization combination of chitosan and bacillus amyloliquefaciens [ P ]. 201410398390.2 ], Liuhai phoenix, Liu Zhen, Yang Rong, an ultrafine powder traditional Chinese medicine for treating tumors [ P ]. ZL201010589748.1 ]. However, because it is insoluble in water, there is a limitation in development and application, and many scholars obtain chitooligosaccharide or chitosan oligosaccharide by degrading chitin or chitosan, and through a series of experiments, it is found that chitooligosaccharide or chitosan oligosaccharide is not only easy to disperse and be absorbed by the body due to good water solubility, but also has various physiological functions, such as antibiosis, antitumor, plant defense capability improvement, etc. Therefore, the further research and development of the functions of chitosan oligosaccharide and chitosan oligosaccharide arouse the attention at home and abroad.

Diabetes mellitus is largely divided into two major groups, type I and type II, which differ mainly in the presence or absence of insulin dependence/resistance. At present, the diabetes patients mainly have type II diabetes, namely non-insulin dependent diabetes mellitus, and account for more than 90 percent of the total number of patients.

The pathogenesis of type II diabetes has not yet been fully elucidated, and defects in insulin resistance and islet beta cell function are the two basic pathological features and the basis of pathogenesis of type II diabetes. The research on the action and the insulin resistance mechanism of the insulin also becomes the main direction for the research on the mechanism of the type II diabetes. Research shows that insulin resistance only appears in obese patients and partial non-obese patients, while patients with type II diabetes have long-term progressive beta cell injury regardless of obesity, beta cell secretion function is gradually degraded due to natural course of disease, and damage to beta cell function can be aggravated by hyperglycemia toxicity, hyperlipidemia toxicity and the like, and then pancreatic function failure (Liu Yonggui, Jie xing, Wujiang, and the like; research progress of new target drugs for treating type 2diabetes [ J ] modern drugs and clinic [ 2015(2): 222-.

At present, sufficient researches report that chitosan oligosaccharide has the functions of reducing blood pressure, blood sugar and blood fat and adsorbing cholesterol, for example, chitosan oligosaccharide can improve and treat diabetes from multiple mechanisms, such as reducing postprandial and fasting blood sugar, improving oral glucose tolerance, improving insulin sensitivity index and improving insulin atrophy of type II diabetic rats; reducing blood lipid and blood pressure, improving lipid metabolism disorder, and improving the body antioxidant capacity of diabetic mice (Rao, L., Ma, Y., Zhuang, M., Luo, T., Wang, Y. and Hong, A., Chitosan-purified selenium as proteins carriers to improve the in vivo clinical of the polypeptide therapeutic BAY 55-9837 for type 2diabetes mellitis, Int J Nanomedicine,2014,9: 4819-4828; Quqiu, Xiaolin, Wang Xiuwu, research on the hypoglycemic effect of Chitosan chelate chromium on diabetic mice. natural products and development, 2012,24(5): oligosaccharide). However, the research on the anti-diabetic function of chitosan oligosaccharide is very limited.

Katiyar et al (Katiyar D, Singh B, Lall A M, et al, effectiveness of chitosan for the management of diabetes in an infected mouse: a synergistic study with anti-hyper and anti-reactive activity [ J ]. Eur J Pharm Sci.2011,44(4): 534; 543) report that a mixture of chitooligosaccharides can reduce fasting plasma glucose and blood lipid levels in a tetraoxypyrimidine-induced type I diabetic mouse and control their serum and hepatopulmonary peroxidation levels, which study is a mixture of oligosaccharides from chitin (N-acetylglucosamine) to chitosan, and the respective oligosaccharide contents therein are not specified. At present, no literature and patent reports on the anti-II diabetes function of chitosan oligosaccharide exist.

Disclosure of Invention

Aiming at the defects in the prior art, the invention aims to provide a novel function of chitobiose and application thereof in health-care food, and experiments show that the chitobiose has the function of remarkably preventing or improving type II diabetes, and has important significance for developing health-care food containing the chitobiose in the future.

In order to achieve the above purposes, the technical scheme adopted by the invention is as follows:

the novel function of chitobiose is characterized in that: the chitobiose has the activity of obviously inhibiting alpha-glucosidase, thereby exerting the efficacy of reducing the blood sugar level of type II diabetes patients.

Another new function of chitobiose is characterized in that: the chitosan disaccharide and the oligosaccharide are compounded, so that the chitosan disaccharide and the oligosaccharide have good blood sugar reducing effect.

On the basis of the technical scheme, the oligosaccharide can be xylooligosaccharide, and the compounding ratio of the chitobiose to the oligosaccharide is 1: 1-1: 4.

Application of chitobiose in preparing health food for treating type II diabetes is provided.

Application of chitobiose and oligosaccharide in preparing health food for type II diabetes is provided.

On the basis of the technical scheme, the chitobiose has the functions of inhibiting the activity of alpha-glucosidase and reducing blood sugar.

On the basis of the technical scheme, the dosage form of the health food is tablets, capsules, oral liquid or sublingual spray.

On the basis of the technical scheme, the mass percent of the chitobiose in the health-care food is 10-20%.

The novel function of the chitobiose is to make sure that the chitobiose can obviously reduce fasting blood glucose, improve oral glucose tolerance and obviously reduce blood lipid level and pancreatic lipid peroxidation level. In addition, the chitobiose can also cooperate with xylooligosaccharide to play a good role in reducing blood sugar.

Detailed Description

The novel functions of chitobiose are: the chitobiose can obviously inhibit the activity of alpha-glucosidase and has good function of reducing blood sugar. Therefore, the chitobiose can be used for preventing or improving type II diabetes, and is proved by the following experimental results:

the chitobiose can obviously reduce fasting blood glucose of diabetic mice, improve oral glucose tolerance, and obviously reduce blood lipid level and pancreatic lipid peroxidation level of the mice;

the chitobiose can cooperate with xylooligosaccharide to play a good role in reducing blood sugar;

after a diabetic (volunteer) takes the health-care product (tablet, capsule, oral liquid and the like) containing the chitobiose for 3 months, the fasting blood sugar of the diabetic is obviously reduced.

The chitobiose used in the invention has high safety and can be developed into health-care food for effectively preventing or improving type II diabetes.

The application of the chitobiose in the health-care food is that according to the method for preparing different health-care foods (functional foods), the chitobiose (accounting for 10-20% by mass percent) is added with ingredients to be prepared into tablets, capsules, oral liquid or sublingual spray. The ingredients (additives) include, but are not limited to: excipient, disintegrant, adhesive and solubilizer. These dosage forms are prepared according to methods well known to those skilled in the art and will not be described in detail.

The invention is further illustrated by the following examples:

example 1: alpha-glucosidase inhibitory Activity of Chitosan disaccharide

An alpha-glucosidase inhibitory activity of chitobiose (prepared enzymatically, Yang S, Fu X, Yan Q, et al cloning, expression, purification and application of a novel protease from a thermophilic marine bacterium, bacterium Paenibacillus, Food Chem,2016,192: 1041-.

Briefly described as follows:

chitobiose with the concentration of 1, 2, 3, 4, 5, 6mg/mL respectively, acarbose with the concentration of 0.1, 0.2, 0.3, 0.4, 0.5, 0.6mg/mL respectively, and alpha-glucosidase solution with the concentration of 0.1U/mL (alpha-glucosidase, EC 3.2.1.20, from Saccharomyces cerevisiae, cat.no. G5003-100UN, Sigma-Aldrich, Inc., St.Louis, MO, USA) are mixed and reacted, then p-nitrophenyl alpha-D-glucoside solution with the concentration of 0.5mmol/L is added as a substrate (4-N-trophenol-alpha-D-glucopyranoside, alpha-pNPG, cat.405. N1377-1G, Sigma-rich, Inc, St.Louis, MO, USA) and the absorbance value is recorded as A nm after the reaction is terminated.

Calculating the inhibition rate of samples with different concentrations on alpha-glucosidase according to the following formula: inhibition ratio (%) ═ a_control–A_sample)/A_controlX 100% where A_controlAbsorbance value of 405nm for blank solution without sample added. The calculation method of IC50 is as follows: the logarithm of the concentration value of the sample with the base 10 was plotted as the abscissa and the inhibition ratio was plotted as the ordinate, and a logarithmic curve of the inhibition ratio with the concentration was plotted to obtain a logarithmic value of the concentration corresponding to a value of y equal to 0.5 (inhibition ratio 50%), and the concentration of the sample was calculated and obtained, i.e., the IC50 value of the sample.

The experimental results are as follows: the IC50 values of the inhibitory action of acarbose and chitobiose on the activity of alpha-glucosidase are 0.26mg/mL and 2.0mg/mL respectively, which indicates that chitobiose has obvious alpha-glucosidase inhibitory activity.

Example 2: preventive and therapeutic effects of chitobiose on type II diabetic mice

One, materials and methods:

1. drugs and reagents

Acarbose, chitobiose, from example 1.

Streptomycin, cat.no. v900890, Sigma-Aldrich, inc., st.louis, MO, usa.

Alloxan, cat.no. a7413, Sigma-Aldrich, inc., st.louis, MO, usa.

2. Animal feeding and administration

ICR (institute of Cancer research) male mice weighing 18-20 g were randomly divided into the following groups:

1. in the healthy control group, the number of patients,

2. the control group of the model was,

3. positive control group (acarbose group),

4. chitobiose group, specifically included three dose groups: low dose 25 mg/kg. d, medium dose 50 mg/kg. d, high dose 100 mg/kg. d,

the above total 6 groups, each group containing 20.

The healthy control group is fed with common feed, and the other groups are fed with high-fat and high-sugar feed. After being fed for 31d, the mice of other groups are subjected to diabetes modeling except for a healthy control group, and the method is shown in three methods for establishing a diabetes mouse model (Zhuxiaoying, Xiaofeng, Liwen, Shijuan, Douging, Huanglingling, Zhang Shu Tu, practical diabetes journal, 2012(03):14-15), and the fasting blood sugar values of the 2d and the 5d are respectively measured, wherein the blood sugar value is higher than 7.8mmol/L and is a success model (the molding rate is more than or equal to 75%);

the healthy control group and the model control group are perfused with normal saline (0.9% NaCl)10 mL/kg.d; positive control group gavage acarbose 100mg/kg d;

a chitobiose low-dose group, a chitobiose medium-dose group and a chitobiose high-dose group, wherein the chitobiose is gavage at 25 mg/kg.d, 50 mg/kg.d and 100 mg/kg.d respectively;

each group of mice was gavaged continuously for 4 weeks, during which time the mice were free to drink and eat food, food intake was recorded daily, body weight was recorded once a week and fasting blood glucose was measured. After the end of the last gavage, the oral glucose tolerance of the mice was determined. Blood is collected from orbit after normal feeding for 3d, serum is separated, and serum triglyceride, total cholesterol and high density lipoprotein cholesterol levels are measured. Mice were sacrificed by cervical dislocation and pancreatic tissue was isolated. Lipid peroxidation levels in pancreatic tissue were determined.

3. Determination of fasting plasma glucose

All group mice had fasting blood glucose measured once a week from the beginning of gavage treatment. Before measurement, fasting is carried out for 12 hours without water supply, blood is taken after tail breaking, and the blood sugar value is measured by a Roche glucometer.

4. Determination of oral glucose tolerance

After the gavage treatment of all the mice of all groups is completed for four weeks, the mice are fasted for 12 hours without water prohibition, the mice are gavaged by 20 percent glucose solution (2g/kg), blood is taken after the tail is broken, and the blood sugar is measured by a Roche glucometer respectively at 0min, 30min, 60min and 120min after the glucose injection. And calculating the area under the curve (AUC) according to the formula:

AUC(mmol.h/L)＝X1/2+X2+X3+X4/2

wherein X1, X2, X3 and X4 represent blood glucose levels of 0, 0.5, 1 and 2h, respectively.

5. Serum lipid metabolism level detection

After the oral glucose tolerance test is normally fed for 3 days, blood is taken from the orbit of the mouse, serum is separated, and the serum Triglyceride (TG) and the Total Cholesterol (TC) are measured by using a full-automatic biochemical analyzer. The content of high density lipoprotein cholesterol (HDL-C) in serum is determined by an enzymatic colorimetric method.

6. Determination of superoxide dismutase and malondialdehyde content in pancreas homogenate supernatant

Pancreatic tissue was homogenized and the levels of Superoxide Dismutase (SOD) and Malondialdehyde (MDA) in the supernatant of the pancreatic homogenate were determined as required by reference to kit instructions (tokyo makino). The protein concentration of the supernatant of the pancreatic tissue homogenate was measured using the Bradford method with bovine serum albumin as a standard solution (Bradford M M.A rapid and sensitive method for the quantification of the microorganisms of protein digestion [ J ]. Anal biochem.1976,72: 248. 254.).

SOD activity is expressed as U/mg prot, and MDA content is expressed as nmol/mg prot.

7. Statistical method

The data results obtained by the test are analyzed by using SAS software to carry out One-Way ANOVA, and Tukey multiple test is used for determining the significance difference between the data, wherein the significance level is p < 0.05.

Second, experimental results

1. Effect of chitobiose on body weight in diabetic mice

Body weights were weighed weekly and recorded during the administration period of the mice, and the results are shown in table 1, with no water deprivation or fasting prior to weighing.

TABLE 1 diabetic mice groups weight cycle Change

As can be seen from the body weight data recorded in Table 1, the body weight of mice in the three dose groups of chitobiose 25, 50 and 100 mg/kg. d remained substantially constant over the course of 4 weeks of administration. The weight of the positive control (acarbose) group slightly increased, which indicates that the weight increase of the mice can be controlled by the chitobiose within the range of the gavage dosage of 25-100 mg/kg-d.

2. Effect of Chitosabiose on fasting plasma glucose in diabetic mice

After the mice are fasted for 12 hours without water supply, the mice are subjected to tail breaking and blood sampling, and the blood sugar value is measured by a Roche glucometer. Blood glucose levels were measured and recorded before and weekly after dosing (see table 2).

TABLE 2 fasting blood glucose level cycle Change in groups of diabetic mice

The blood glucose levels in the positive control (acarbose) group were decreased by comparison of the fasting blood glucose levels before administration and before dissection in Table 2. Secondly, the blood sugar value of each dose group of the chitobiose is obviously reduced, wherein the blood sugar reduction effect of the dose group of 100 mg/kg.d is most obvious, the blood sugar reduction degree is better than that of a positive control group, and the blood sugar reduction degree is close to the upper limit of the fasting blood sugar range of a normal mouse (the fasting blood sugar of the normal mouse is less than 6.1 mmol/L).

3. Effect of Chitosabiose on oral glucose tolerance in diabetic mice

After the administration period of each group of diabetic mice was completed, fasting was not performed for 12 hours, oral glucose tolerance test was performed, and blood glucose values at 0min, 30min, 60min and 120min after gastric gavage were measured and recorded (see table 3).

TABLE 3 blood glucose value variation in oral glucose tolerance test of each group of diabetic mice

From the results of the experiment shown in Table 3, it was found that the blood glucose level of each group of mice increased 30min after the administration of the gastric glucose solution, and then gradually decreased with the lapse of time. The area under the curve (AUC) of the chitobiose 50 and 100 mg/kg.d dose groups is smaller than that of the acarbose of the positive control group, which indicates that the body regulates the sugar metabolism level better than that of the acarbose group.

4. Effect of chitobiose on serum lipid metabolism in diabetic mice

Blood was collected from the orbit of the mouse, and serum was separated to measure the contents of Triglyceride (TG), Total Cholesterol (TC) and high density lipoprotein cholesterol (HDL-C) in the serum (see Table 4).

As can be seen from Table 4, the normal group and the model group have significant differences in TG, CHO and HDL-C contents, the TG and CHO contents of the model group are significantly greater than those of the normal group, and the HDL-C content is less than that of the normal group, so that the lipid metabolism disorder characteristics of the diabetic mice are met. The lipid-lowering effect of each group of chitobiose was mainly shown by lowering serum total Triglyceride (TG) level, and then the serum HDL-C content was significantly increased compared with that of the model group, while the serum total Cholesterol (CHO) level of each group was not significantly different from that of the model group except for the 100 mg/kg-d dose group. In each dose group of chitobiose, 100 mg/kg. d dose groups were close to normal mice in TG and CHO, therefore, 100 mg/kg. d is the best dose of chitobiose for treating diabetic mice.

TABLE 4 serum lipid metabolism levels in groups of diabetic mice

5. Effect of chitobiose on pancreatic lipid peroxidation levels in diabetic mice

Mouse pancreatic lipid peroxidation levels are shown in table 5.

TABLE 5 lipid peroxidation levels of pancreatic glands in groups of diabetic mice

Note: "mg prot" in a unit means the unit of T-SOD activity (U) or the amount of MDA (nmol) per mg protein.

As shown in Table 5, the activity of SOD in the pancreas was greatly reduced as compared with that of normal mice in the diabetic model, which was about 40% of that of healthy mice. The production of lipid peroxidation product MDA is obviously increased and is about 2 times of that of normal mice. Through the gastric lavage treatment, the activity of the positive control acarbose group SOD is obviously improved, and compared with a model group containing MDA, the positive control acarbose group SOD has no obvious difference. In each dose group of the chitobiose, SOD activity is greatly increased in 50 mg/kg-d dose groups and SOD content is obviously reduced, which shows that the effect of reducing lipid peroxidation level is better than that of acarbose group.

Example 3: hypoglycemic effect compounded by chitobiose and xylo-oligosaccharide

The diabetic mice were modeled as described in example 2. The complex formulation of chitobiose and xylooligosaccharide (Biochemical reagent, Shandong dragon biological science and technology Co., Ltd.) comprises three groups: chitosan disaccharide (25, 50, 100 mg/kg. d) with three doses is respectively compounded with xylo-oligosaccharide (100 mg/kg. d) (compounding ratio: 1:4, 1:2, 1:1), the xylo-oligosaccharide (100 mg/kg. d) and the chitobiose (100 mg/kg. d) are used as controls, and a healthy control group, a model control group and a positive control group (acarbose group) are simultaneously established, wherein the total number of the groups is 8, and each group is 20.

The mice of each group were gavaged continuously for 4 weeks, during which time the mice were free to drink and eat food, food intake was recorded daily, body weight was recorded once a week and fasting blood glucose was measured. By comparing the fasting blood glucose values before and after the gavage treatment, the blood glucose values of the positive control group and each gavage treatment group are obviously reduced except that the fasting blood glucose of the mice of the model control group is not obviously changed (10.1-11.3 mmol/L).

When the xylo-oligosaccharide (100, 200 and 400mg/kg multiplied by d) is used for gastric perfusion alone, the fasting blood glucose of the mice is reduced by 17 percent (from 10.2mmol/L to 8.4mmol/L), and after the xylo-oligosaccharide is compounded with the chitobiose (50mg/kg multiplied by d), the fasting blood glucose of the treated type II diabetes mice is reduced from 10.5mmol/L to 7.2mmol/L and is reduced by 31 percent, which shows that the chitobiose and the xylo-oligosaccharide have better blood glucose reducing effect after being compounded, and the respective treatment dose of the chitobiose and the oligosaccharide can be reduced.

Example 4: tablet and capsule preparation

According to the specific meter of the body surface area of the human body and the animal body in Xutaiyun pharmacological experiment methodology, the oral dose of 769.2mg/d, which is the therapeutic dose, of the mouse gavage dose of 100mg/kg d relative to the human body (taking 70kg of body weight as an example) can be obtained through conversion. Examples 4-5 are all treated orally at this dose.

Tablets and capsules of chitobiose were prepared with reference to the tablet and capsule process of acarbose.

The tablet formula comprises: 100g of chitobiose, 85g of microcrystalline cellulose, 19g of sodium carboxymethyl starch, 20g of low-substituted hydroxypropyl cellulose, 36g of crospovidone, 1g of micropowder silica gel and 0.5g of magnesium stearate, and 1000 tablets are prepared (Xiexiang, Liaoxiangru, Cheng, and the like. powder direct tabletting method is used for preparing acarbose tablets [ J ]. Chinese pharmacist, 2011,14(9): 1276-one 1278, Liuliu Xiongyang, Xiexiang, Zhongjian. preparation and dissolution rate determination [ J ]. Chinese pharmacist, 2011,14(11): 1612-one 1614, Sun Heying, Chenchunkui, Duckweed, acarbose sustained-release tablets [ J ]. Hebei institute of Industrial medicine, 2008,25(1): 12-14.).

Wherein microcrystalline cellulose is used as filler, sodium carboxymethyl starch, low-substituted hydroxypropyl cellulose and crospovidone peppermint oil are used as disintegrating agents, aerosil is used as glidant, and magnesium stearate is used as lubricant. Each tablet contains 100mg of chitobiose. The oral preparation is orally taken by adults (70kg of body weight) three times a day, and 2-3 tablets are taken each time.

The capsule formula comprises: 200g of chitobiose, 20g of micropowder silica gel, 380g of dextrin and 400g of starch, which are fully mixed and encapsulated into 1000 capsules (Pengyong pine, Weichengzhi, content of acarbose capsules and related substances [ J ] determined by HPLC, journal of Waisi pharmacy, 2004(01): 58-59; Wangjingfeng, Hushiwei, Changliang, and the like.

Each capsule contains 1g of chitobiose 200mg (20%). The oral preparation is orally taken by an adult (70kg of body weight) twice a day, and 1-2 granules are taken each time.

10-15 type II diabetic patients and obese healthy volunteers (without clinical symptoms of type II diabetes) were collected, oral chitobiose tablets were administered at regular intervals according to the dose of the chitobiose tablet in example 4, fasting blood glucose was measured every week, and blood glucose changes were measured within 3 months. (Lvskitin, an oral liquid and sublingual spray for treating diabetes [ P ].2014101501055.2014.08.27 ], Dingzheng, Shijinsong, Sundafeng, etc., a dietary formula containing chitosan oligosaccharide Dunaliella salina powder for diabetic patients [ P ].201510179903.5.2015.07.15 ].

The change result of fasting blood glucose of each volunteer within 3 months is monitored, and the chitobiose reduces the fasting blood glucose value of the type II diabetes patients to different degrees (from 11.3-15.6 mmol/L to 7.5-8.2 mmol/L after treatment); in addition, the fasting blood sugar of the obese healthy subject is reduced to 6.5-7.3 mmol/L after treatment from 7.1-8.3 mmol/L before the chitosan tablet is orally taken.

Example 5: oral liquid and sublingual spray

The formula of the oral liquid and the sublingual spray comprises: 20g of chitobiose and a proper amount of honey, adding distilled water to 1000mL, uniformly mixing, sterilizing and filling, wherein each 10mL contains 200mg of chitobiose.

The oral liquid is orally taken by an adult (70kg of body weight) twice a day, and 1-2 pills are taken each time. The sublingual spray is filled into a spray bottle, and is sprayed 3 times to 6 times per day (Lvskitin, an oral liquid for treating diabetes and sublingual spray [ P ].2014101501055.2014.08.27 ]). The two solutions can be used alone or in combination to enhance therapeutic effect (gastrointestinal tract and oral mucosa absorption).

In example 4, fasting glucose changes were monitored in 10-12 type II diabetics and obese healthy volunteers 3 months before and after oral liquid treatment. The result shows that the chitobiose oral liquid also has obvious function of reducing blood sugar.

The invention discloses a function of chitobiose in preventing or improving type II diabetes and application thereof in health-care food. The foregoing is a representative embodiment of the present invention, and is not intended to limit the present invention in any way, and any person skilled in the art may make changes or modifications to the equivalent embodiment using the above-identified matters. However, any simple modification, equivalent change and modification of the above embodiments according to the technical essence of the present invention will still fall within the protection scope of the technical solution of the present invention without departing from the technical solution of the present invention.

Those not described in detail in this specification are within the skill of the art.

Claims

1. The application of the chitobiose in preparing the health food for type II diabetes is characterized in that: the chitobiose and xylooligosaccharide are compounded for reducing blood sugar, and the compounding ratio of the chitobiose to the xylooligosaccharide is 1: 1-1: 4.

2. The use of chitobiose of claim 1 in the preparation of a health food for type II diabetes, wherein: the chitobiose has effects of inhibiting alpha-glucosidase activity and reducing blood sugar.

3. The use of chitobiose of claim 1 in the preparation of a health food for type II diabetes, wherein: the dosage form of the health food is tablets, capsules, oral liquid or sublingual spray;

the health food comprises 10-20% of chitobiose by mass.