Collagen peptide composition for regulating metabolism and losing weight and fat and application thereof

Abstract

The application discloses a collagen peptide composition for regulating metabolism and reducing weight and fat, and application thereof. The composition comprises collagen peptide fermentation liquor and plant enzyme, and the volume ratio of the collagen peptide fermentation liquor and the plant enzyme is 10-15:2-5. The collagen peptide fermentation liquor is obtained by fermenting collagen peptide by Lactobacillus rhamnosus NCU2217. The plant enzyme is obtained by fermenting a plurality of traditional Chinese medicine compositions by Lactobacillus casei. The collagen peptide fermentation liquor is obtained by fermenting collagen peptide by Lactobacillus rhamnosus NCU2217, and the plant enzyme is obtained by fermenting ganoderma lucidum, alisma and medicated leaven in cooperation. Only in this way, the collagen peptide fermentation liquor and the plant enzyme obtained in cooperation have more remarkable effects of regulating lipid metabolism, reducing weight and fat, reducing blood sugar and protecting liver damage.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of biological medicine, and particularly relates to a collagen peptide composition for regulating metabolism and reducing weight and fat and application thereof. BACKGROUND

[0002] With the improvement of life quality, hyperglycemia, hyperlipidemia, obesity and the like gradually threaten human health and show a trend of youth, which has seriously affected the life quality of people. At present, the treatment of obesity, hyperglycemia and hyperlipidemia mainly relies on western medicine, which has limited efficacy and obvious side effects after long-term administration. In view of this situation, the development of safe and effective lipid-lowering, hypoglycemic and weight-reducing drugs has become the focus of current medical research.

[0003] Compared with western medicine, traditional Chinese medicine has the advantages of safety, fewer side effects, no damage to the gastrointestinal tract and little stimulation to the liver and kidney, and thus attracts people's attention. However, traditional Chinese medicine has the problems of slow effect and inconvenient administration. The present application is based on collagen peptide and traditional Chinese medicine, supplemented by biological fermentation technology, and develops a collagen peptide composition which has a significant effect on weight loss, fat reduction, blood glucose reduction and lipid metabolism regulation. SUMMARY

[0004] The present application aims to overcome the deficiencies of the prior art and provide a collagen peptide composition for regulating metabolism and reducing weight and fat and application thereof, so as to solve the problems in the above technical background.

[0005] Chinese patent 201810978527.X (applicant: Nanchang University) discloses lactobacillus rhamnosus NCU2217, which has been preserved in China General Microbiological Culture Collection Center (CGMCC) on April 9, 2018, and the preservation number is CGMCC NO 15573. The lactobacillus rhamnosus NCU2217 used in the present application is from the researchers of Nanchang University.

[0006] In the research and practice process, the team of the present application found that the collagen peptide fermentation liquor obtained by fermentation of lactobacillus rhamnosus NCU2217 and plant enzyme have a significant effect of regulating sugar / lipid metabolism, reducing weight and fat and reducing blood glucose in cooperation. However, the collagen peptide fermentation liquor obtained by fermentation of collagen peptide without fermentation or ordinary lactobacillus rhamnosus and plant enzyme has far insufficient effects of regulating sugar / lipid metabolism, reducing weight and fat, reducing blood glucose and protecting liver damage.

[0007] To achieve the above-mentioned purpose, the present application realizes the following technical scheme:

[0008] In a first aspect, the present application provides a collagen peptide composition for regulating metabolism and reducing weight and fat, comprising a collagen peptide fermentation liquor and a plant enzyme, wherein the volume ratio of the collagen peptide fermentation liquor and the plant enzyme is 10-15:2-5; the collagen peptide fermentation liquor is obtained by fermenting collagen peptide with Lactobacillus rhamnosus; and the plant enzyme is obtained by fermenting a traditional Chinese medicine composition with Lactobacillus casei.

[0009] In the above technical solution, the volume ratio of the collagen peptide fermentation liquor and the plant enzyme is 12:3.

[0010] In the above technical solution, the specific fermentation method of the collagen peptide fermentation liquor is as follows: collagen peptide 3-5% (w / v) is added to water, and Lactobacillus rhamnosus is inoculated, and then fermentation is carried out at 30-42°C for 16-24h to obtain the collagen peptide fermentation liquor.

[0011] In the above technical solution, the molecular weight of the collagen peptide is 2000-6000 Dalton; preferably, the molecular weight of the collagen peptide is 2000-3000 Dalton. Specifically, the collagen peptide is any one of fish collagen peptide, bovine collagen peptide and pig collagen peptide. The main raw material of the fish collagen peptide is fish skin; for example, fish collagen peptide with a molecular weight of 2000 Da is purchased from Hebei Saiyi Biological Technology Co., Ltd.; bovine collagen peptide with a molecular weight of 2000-4000 Da is purchased from Shaanxi Chenming Biological Technology Co., Ltd.; and pig collagen peptide with a molecular weight of 3000 Da is purchased from Shanghai Shucan Industry Co., Ltd.

[0012] In the above technical solution, the Lactobacillus rhamnosus is Lactobacillus rhamnosus NCU2217, which is preserved in the China General Microbiological Culture Collection Center with a preservation number of CGMCC NO 15573.

[0013] In the above technical solution, the inoculation amount of the Lactobacillus rhamnosus is 1×10 6 ~1×10 7 cfu / mL.

[0014] In the above technical solution, the traditional Chinese medicine composition comprises Ganoderma lucidum, Alisma orientalis and Shenqu.

[0015] In the above technical solution, the weight ratio of Ganoderma lucidum, Alisma orientalis and Shenqu is 3:2:2.

[0016] In the above technical solution, the specific preparation method of the plant enzyme is as follows:

[0017] (a) The traditional Chinese medicine materials in the traditional Chinese medicine composition are mixed according to the proportion, and then added to distilled water for extraction. The obtained extract is concentrated to obtain a traditional Chinese medicine concentrate;

[0018] (b) inoculating Lactobacillus casei directly into the Chinese medicine concentrate to perform inoculation fermentation, and obtaining plant enzyme.

[0019] In the technical solution, the inoculation amount of the Lactobacillus casei is 1×10 6 ~1×10 7 cfu / mL.

[0020] In the second aspect, the application further provides application of the collagen peptide composition described above in the preparation of a medicine.

[0021] Further, the medicine is a medicine with the effects of regulating metabolism, reducing weight and fat, and protecting liver damage.

[0022] Compared with the prior art, the application has the following beneficial effects:

[0023] 1. The collagen peptide fermentation liquor and the plant enzyme in the application have the effects of reducing weight and fat, reducing blood sugar, regulating sugar metabolism, lipid metabolism, and protecting liver damage.

[0024] 2. The collagen peptide fermentation liquor in the application is obtained by fermentation of Lactobacillus rhamnosus NCU2217, and the plant enzyme is obtained by synergistic fermentation of Ganoderma lucidum, Alisma orientale and Radix Gastrodiae. Only the collagen peptide fermentation liquor and the plant enzyme obtained by the above method have more remarkable effects of regulating sugar / lipid metabolism, reducing weight and fat, reducing blood sugar, and protecting liver damage. BRIEF DESCRIPTION OF DRAWINGS

[0025] Figure 1 A curve graph of the influence of the collagen peptide composition in the application on the weight of mice;

[0026] Figure 2 A columnar graph of the influence of the collagen peptide composition in the application on the content of TG, TC, LDL-C, HDL-C and LEP in the serum of mice;

[0027] Figure 3 A columnar graph of the influence of the collagen peptide composition in the application on the glycogen content in the liver of rats;

[0028] Figure 4 A columnar graph of the influence of the collagen peptide composition in the application on the content of ALT and AST in the serum of mice;

[0029] Figure 5 A columnar graph of the influence of the collagen peptide composition in the application on the content of SOD and MDA in the liver of mice. DETAILED DESCRIPTION

[0030] Following specific examples are used to illustrate the embodiments of the present application, and other advantages and effects of the present application can be easily understood by those skilled in the art from the disclosure of this specification. The present application can also be implemented or applied by other different embodiments, and various modifications or changes can be made to the details in this specification based on different views and applications without departing from the spirit of the present application. It should be noted that the following examples and features in the examples can be combined with each other without conflict.

[0031] Example 1

[0032] The present example provides a collagen peptide composition for regulating metabolism and reducing weight and fat, comprising a collagen peptide fermentation liquor and a plant enzyme, and the volume ratio of the collagen peptide fermentation liquor and the plant enzyme is 10:5; the collagen peptide fermentation liquor is obtained by fermenting collagen peptide with Lactobacillus rhamnosus; and the plant enzyme is obtained by fermenting a traditional Chinese medicine composition with Lactobacillus casei. The traditional Chinese medicine composition comprises Ganoderma lucidum 3 parts, Alisma orientale 2 parts and Radix Gastrodiae 2 parts by weight. The collagen peptide used is fish collagen peptide. The fish collagen peptide has a molecular weight of 2000 Da and is purchased from Hebei Saiyi Biological Technology Co., Ltd.

[0033] In the present example, the specific fermentation method of the collagen peptide fermentation liquor is as follows: collagen peptide 5% (w / v) is added to water, and Lactobacillus rhamnosus is inoculated, and then the mixture is fermented at 30°C for 24 h to obtain the collagen peptide fermentation liquor. The Lactobacillus rhamnosus is Lactobacillus rhamnosus NCU2217, which is preserved in the China General Microbiological Culture Collection Center, and the preservation number is CGMCC NO 15573; and the inoculation amount of the Lactobacillus rhamnosus is 1×10 6 cfu / mL.

[0034] In the present example, the specific preparation method of the plant enzyme is as follows:

[0035] (a) The traditional Chinese medicine materials in the traditional Chinese medicine composition are mixed in proportion, and then added to 10 times the amount of distilled water for extraction. After 3 times of extraction, the filtrates are combined, and the obtained extract is concentrated to a crude drug concentration of 1.0 g / ml to obtain a traditional Chinese medicine concentrate;

[0036] (b) Lactobacillus casei is directly inoculated into the traditional Chinese medicine concentrate for inoculation and fermentation, and then the mixture is fermented at 37°C for 4 h to obtain the plant enzyme. The Lactobacillus casei is Lactobacillus casei ATCC 393, order number 0176P, purchased from Shenzhen Zike Biological Technology Co., Ltd.; and the inoculation amount of the Lactobacillus casei is 2×10 6 cfu / mL.

[0037] Example 2

[0038] The embodiment provides a collagen peptide composition for regulating metabolism and reducing weight and fat, which comprises a collagen peptide fermentation liquor and a plant enzyme, and the volume ratio of the collagen peptide fermentation liquor and the plant enzyme is 12:3; the collagen peptide fermentation liquor is obtained by fermenting collagen peptide with Lactobacillus rhamnosus NCU2217; and the plant enzyme is obtained by fermenting a plurality of traditional Chinese medicine compositions with Lactobacillus casei. The traditional Chinese medicine compositions contain Ganoderma lucidum 3 parts, Alisma orientale 2 parts and Radix Ginseng 2 parts in terms of weight fraction. The collagen peptide is fish collagen peptide, which has a molecular weight of 2000 Da and is purchased from Hebei Saiyi Biotechnology Co., Ltd.

[0039] In the embodiment, the specific fermentation method of the collagen peptide fermentation liquor is as follows: collagen peptide 4% (w / v) is added to water, and Lactobacillus rhamnosus is inoculated, and then fermentation is carried out at 37 ℃ for 20 h to obtain the collagen peptide fermentation liquor. The Lactobacillus rhamnosus is Lactobacillus rhamnosus NCU2217, which is preserved in the China General Microbiological Culture Collection Center, and the preservation number is CGMCC NO 15573; and the inoculation amount of the Lactobacillus rhamnosus is 3×10 6 cfu / mL.

[0040] In the embodiment, the specific preparation method of the plant enzyme is as follows:

[0041] (a) the traditional Chinese medicine materials of the traditional Chinese medicine composition are mixed according to the proportion, and then added to 10 times the amount of distilled water for extraction, and the filtrates are combined after three times of extraction, and the obtained extraction liquid is concentrated to a crude drug concentration of 1.0 g / ml to obtain a traditional Chinese medicine concentrated solution;

[0042] (b) Lactobacillus casei is directly inoculated and fermented in the traditional Chinese medicine concentrated solution, and then fermentation is carried out at 37 ℃ for 4 h to obtain the plant enzyme. The Lactobacillus casei is Lactobacillus casei ATCC 393, order number 0176P, purchased from Shenzhen Zike Biological Technology Co., Ltd.; and the inoculation amount of the Lactobacillus casei is 1×10 6 cfu / mL.

[0043] Embodiment 3

[0044] The embodiment provides a collagen peptide composition for regulating metabolism and reducing weight and fat, which comprises a collagen peptide fermentation liquor and a plant enzyme, and the volume ratio of the collagen peptide fermentation liquor and the plant enzyme is 15:2; the collagen peptide fermentation liquor is obtained by fermenting collagen peptide with Lactobacillus rhamnosus; and the plant enzyme is obtained by fermenting a plurality of traditional Chinese medicine compositions with Lactobacillus casei. The traditional Chinese medicine compositions contain Ganoderma lucidum 3 parts, Alisma orientale 2 parts and Radix Ginseng 2 parts in terms of weight fraction. The collagen peptide is fish collagen peptide, which has a molecular weight of 2000 Da and is purchased from Hebei Saiyi Biotechnology Co., Ltd.

[0045] In this embodiment, the specific fermentation method of the collagen peptide fermentation liquor is as follows: 3% (w / v) of collagen peptide is added to water, and Lactobacillus rhamnosus is inoculated, and then the mixture is fermented at 42°C for 16 h to obtain the collagen peptide fermentation liquor. The Lactobacillus rhamnosus is Lactobacillus rhamnosus NCU2217, which is preserved in the China General Microbiological Culture Collection Center, and the preservation number is CGMCC NO 15573; the inoculation amount of the Lactobacillus rhamnosus is 1×10 7 cfu / mL.

[0046] In this embodiment, the specific preparation method of the plant enzyme is as follows:

[0047] (a) The traditional Chinese medicine materials of the traditional Chinese medicine composition are mixed according to the proportion, and then added to 10 times of distilled water for extraction. After 3 times of extraction, the filtrates are combined, and the obtained extract is concentrated to a crude drug concentration of 1.0 g / ml to obtain a traditional Chinese medicine concentrate;

[0048] (b) Lactobacillus casei is directly inoculated into the traditional Chinese medicine concentrate for inoculation and fermentation, and then the mixture is fermented at 37°C for 4 h to obtain the plant enzyme. The Lactobacillus casei is Lactobacillus casei ATCC 393, order number 0176P, which is purchased from Shenzhen Zike Biological Technology Co., Ltd.; the inoculation amount of the Lactobacillus casei is 1×10 7 cfu / mL.

[0049] Example 4

[0050] This embodiment is similar to example 2, and the difference is that the collagen peptide is replaced by bovine collagen peptide. The bovine collagen peptide has a molecular weight of 2000-4000 Da, which is purchased from Shaanxi Chenming Biological Technology Co., Ltd.

[0051] Example 5

[0052] This embodiment is similar to example 2, and the difference is that the collagen peptide is replaced by pigskin collagen peptide. The pigskin collagen peptide has a molecular weight of about 3000 Da, which is purchased from Shanghai Shucan Industry Co., Ltd.

[0053] Comparative example 1

[0054] This embodiment is similar to example 2, and the difference is that the collagen peptide fermentation liquor is replaced by Lactobacillus rhamnosus NCU2217 bacterial liquid, and the concentration of Lactobacillus rhamnosus NCU2217 in the Lactobacillus rhamnosus NCU2217 bacterial liquid is 1×10 7 cfu / mL.

[0055] Comparative example 2

[0056] This embodiment is similar to example 2, and the difference is that the collagen peptide fermentation liquor is replaced by 4% (w / v) collagen peptide aqueous solution.

[0057] Comparative Example 3

[0058] This example is similar to Example 2, except that the fermentation bacteria used in the collagen peptide fermentation broth is different, and Lactobacillus rhamnosus NCU2217 is replaced by Lactobacillus rhamnosus, which is Lactobacillus rhamnosus ATCC53103 purchased from Shenzhen Zike Biological Technology Co., Ltd.

[0059] Comparative Example 4

[0060] This example is similar to Example 2, except that the traditional Chinese medicine composition used in the plant enzyme is directly fermented without adding Lactobacillus casei. Specifically, the specific preparation method of the plant enzyme is as follows:

[0061] (a) The traditional Chinese medicine materials of the traditional Chinese medicine composition are mixed according to the proportion, then added to 10 times of distilled water for extraction, and after 3 times of extraction, the filtrate is combined, and the obtained extraction liquid is concentrated to a crude drug concentration of 1.0 g / ml, to obtain a traditional Chinese medicine concentrate;

[0062] (b) The traditional Chinese medicine concentrate is fermented at 37°C for 4h to obtain a plant enzyme.

[0063] Comparative Example 5

[0064] This example is similar to Example 2, except that the traditional Chinese medicine composition used in the plant enzyme removes Ganoderma lucidum.

[0065] Comparative Example 6

[0066] This example is similar to Example 2, except that the traditional Chinese medicine composition used in the plant enzyme removes Alisma orientalis.

[0067] Comparative Example 7

[0068] This example is similar to Example 2, except that the traditional Chinese medicine composition used in the plant enzyme removes Shenqu.

[0069] I. Functionality research of collagen peptide composition for weight loss and regulation of lipid metabolism

[0070] 1.1 Construction of obese mouse model

[0071] Mice: Kunming mice, male, body weight 20±2g, purchased from Jiangsu Ailinfeng Biological Technology Co., Ltd.

[0072] Construction method: all experimental mice were fed with basic feed, and after adaptive feeding for 1 week, they were randomly divided into basic feed group (6) and high-fat feed group (150). The basic feed, also called SPF level mouse maintenance feed, was purchased from Liaoning Changsheng Biotechnology Co., Ltd.; the high-fat feed formula: basic feed 79.5%, lard 10%, egg yolk powder 10%, cholesterol 0.5%. The basic feed group as the normal group continued to be fed with basic feed, and the high-fat feed group as the model group was fed with high-fat feed. The mice were weighed once a week during the modeling period, and the change of body weight of the mice was observed during each week of feeding. When the obesity degree was more than 20%, the obese mouse model was successfully established.

[0073]

[0074] 1.2 Test grouping and administration

[0075] From the obese mice successfully modeled, 96 were randomly divided into 16 groups according to body weight, 1 model group, 1 positive control group, 14 treatment groups (Examples 1-5, Comparative Examples 1-7 groups, collagen peptide fermentation liquor and plant enzyme), to ensure that there is no significant difference in body weight between groups. The basic feed group is the normal group. After grouping, enter the experimental period, the experimental period is 6 weeks, the model group, the positive control group and the treatment group continue to be fed with high-fat feed, the blank group continues to be fed with basic feed, and the food intake is recorded every day. The experimental mice were given different doses of test samples by gavage at 8:30 am every day. The normal group and the model group were given physiological saline 10 mL·Kg -1 (bw / d); the positive control group was given orlistat 54.6 mg·Kg -1 (bw / d); the treatment group was given the composition obtained in Examples 1-5 and Comparative Examples 1-7, respectively, diluted 10 times with distilled water, and given by gavage at a dose of 10 mL·Kg -1 (bw / d). The collagen peptide fermentation liquor and plant enzyme obtained in Example 2 were diluted according to the amount in Example 2 (collagen peptide fermentation liquor 10*12 / 15 times dilution, plant enzyme 10*3 / 15 times dilution), and given by gavage at a dose of 10 mL·Kg -1 (bw / d).

[0076] 1.3 Detection index

[0077] After the experimental period, the mice were fasted for more than 12 h the day before being killed, and the water was not controlled during the fasting period. The body weight and body length of the mice were measured before being killed, and the mice were anesthetized, blood was taken from the eye, and then the mice were decapitated; the testes and perirenal fat of the mice were obtained by laparotomy.

[0078] 1.31 Body weight: each group of mice free to eat and drink water, gavage period each week weighing, continuous 6 weeks, statistical results are shown in Table 1 and Figure 1

[0079] 1.32 Lee's index: at the end of the experiment, record the body weight (g) of the mice before death, measure the body length (cm), the body length is the distance from the tip of the nose to the anus before dissection, calculate Lee's index according to the following formula, the calculation results are shown in Table 2;

[0080]

[0081] 1.33 Fat coefficient: after the mice were killed, the testes and the fat around the kidneys were taken out from the abdominal cavity of the mice and placed in pre-cooled physiological saline. After removing the blood and impurities and absorbing with filter paper, the weight was measured, and the fat coefficient of the mice was measured, that is, the total fat mass (g) of testis fat and kidney fat compared with the body weight (g). The fat coefficient was calculated according to the following formula, and the calculation results are shown in Table 2.

[0082]

[0083] 1.34 After the last administration, the mice were fasted for 12h, and then enucleated and bled after anesthesia. The centrifuge tube containing the mouse blood was placed in the centrifuge, set at 3500r / min speed, and centrifuged at 4℃ for 15min. The supernatant was aspirated for testing the content of triglyceride (TG), cholesterol (TC), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), and leptin (LEP) in the serum of the mice, and the results are shown in Table 3 and Figure 2 The TG, TC, LDL-C, HDL-C, and LEP kits were purchased from Nanjing Jiancheng Biological Engineering Research Institute Co., Ltd.

[0084] Table 1

[0085]

[0086] Table 1 and Figure 1 ​It can be seen that the obesity degree of the model group mice is greater than 20% before gavage, indicating that the obese mouse model is successfully constructed; the body weight of the normal group mice has always shown a growth trend, and the body weight of the example 1-3 and the positive control group is significantly reduced compared with the model group at the 4th and 6th weeks. Compared with the collagen peptide fermentation liquor and the plant enzyme group, the body weight of example 2 is significantly reduced, indicating that the synergistic effect of the collagen peptide fermentation liquor and the plant enzyme on the body weight of the mice plays a role in weight loss, and the effect is better than that of using them alone; compared with the comparative examples 1-7, the body weight of example 2 is significantly reduced, indicating that the collagen peptide fermentation liquor and the plant enzyme together play a role in weight loss of the mice, and only the collagen peptide fermentation liquor fermented by lactobacillus rhamnosus NCU2217 and the plant enzyme fermented by ganoderma lucidum, alisma and radix ginseng are used, and the synergistic effect of the collagen peptide fermentation liquor and the plant enzyme is more significant.

[0087] Table 2

[0088]

[0089] As can be seen from Table 2, compared with the model group, the Lee's index and the fat coefficient of examples 1-3 and the positive control group are significantly reduced. Compared with the collagen peptide fermentation liquor and the plant enzyme group, the Lee's index and the fat coefficient of example 2 are significantly reduced, indicating that the synergistic effect of the collagen peptide fermentation liquor and the plant enzyme can significantly reduce visceral adiposity, and the effect is more obvious when they are used together than when they are used alone; compared with the comparative examples 1-7, the Lee's index and the fat coefficient of example 2 are significantly reduced, indicating that the collagen peptide fermentation liquor and the plant enzyme together can reduce visceral adiposity, and only the collagen peptide fermentation liquor fermented by lactobacillus rhamnosus NCU2217 and the plant enzyme fermented by ganoderma lucidum, alisma and radix ginseng are used, and the synergistic effect of the collagen peptide fermentation liquor and the plant enzyme is more significant.

[0090] As can be seen from the above, the collagen peptide composition obtained by the present application has a significant weight loss and fat loss effect by the synergistic effect of the collagen peptide fermentation liquor and the plant enzyme.

[0091] Table 3

[0092]

[0093] As can be seen from Table 3 and Figure 2It can be known that, compared with the normal group, the TG, TC, LDL-C and LEP of the model group were significantly increased, and the HLDL-C was significantly decreased; compared with the model group, the TG, TC, LDL-C and LEP of the example 1-3 and the positive control group were significantly increased, and the HLDL-C was significantly decreased. Compared with the collagen peptide fermentation liquor and the plant enzyme group, the TG, TC, LDL-C and LEP of the example 2 were significantly increased, and the HLDL-C was significantly decreased, which indicated that the synergistic effect of the collagen peptide fermentation liquor and the plant enzyme had the effect of regulating lipid metabolism and improving the disorder of lipid metabolism of the body, and the effect was more obvious when they were used together; compared with the comparative example 1-7, the TG, TC, LDL-C and LEP of the example 2 were significantly increased, and the HLDL-C was significantly decreased, which indicated that the collagen peptide fermentation liquor and the plant enzyme together could regulate lipid metabolism, and only the collagen peptide fermentation liquor fermented by Lactobacillus rhamnosus NCU2217 and the plant enzyme synergistically fermented by Ganoderma lucidum, Alisma orientale and Radix Menispermi could have more significant effect of regulating lipid metabolism. Therefore, the collagen peptide composition obtained by the present application has the effect of regulating lipid metabolism and improving the disorder of lipid metabolism of the body by the synergistic effect of the collagen peptide fermentation liquor and the plant enzyme.

[0094] II. Function research of collagen peptide composition on regulating sugar metabolism

[0095] 2.1 Grouping and administration: After the purchased SD rats (purchased from Hunan Slaik Jingda Experimental Animal Co., Ltd.) were fed with sterile laboratory general feed for 6 days, all the rats were weighed, fasted for 4 h, and blood was taken from the tail vein, and the fasting blood glucose was measured, then the rats were intragastrically administered with 250 mg / kg of glucose, and the blood glucose values at 0.5 h and 2 h were measured. 90 rats with normal blood glucose were randomly divided into 15 groups according to the blood glucose value, which were 1 blank group, 1 model group, 1 positive control group and 12 treatment groups (example 2, examples 4-5, comparative examples 1-7, collagen peptide fermentation liquor and plant enzyme). The blank group was not treated, the model group was administered with physiological saline at a dose of 10 mL·kg -1 (bw / d); the positive control group was administered with metformin at a dose of 107 mg·kg -1 (bw / d), and the treatment groups were administered with the compositions obtained in examples 2, examples 4-5 and comparative examples 1-7, which were diluted 10 times with distilled water, and then administered at a dose of 10 mL·kg -1 (bw / d). 2.2 Measurement of blood glucose value: the blood glucose value was measured at 0.5 h and 2 h after the administration. -1(bw / d) dose gavage. Collagen peptide fermentation broth and plant enzyme are obtained in Example 2, which are diluted according to the amount of Example 2 (collagen peptide fermentation broth 10*12 / 15 times dilution, plant enzyme 10*3 / 15 times dilution), respectively, 10 mL·kg -1 (bw / d) dose gavage.

[0096] Continuous gavage for 33 days. From the 1st day to the 7th day of gavage, SD rats were fed with ordinary feed, and from the 8th day, rats except the blank group were fed with high-energy feed for 26 days. Ordinary feed, also known as SPF level large mouse maintenance feed, was purchased from Liaoning Changsheng Biotechnology Co., Ltd.; High-energy feed formula: ordinary feed 52.6%, lard 10%, white sugar 15%, protein powder 15%, casein 5%, cholesterol 1.2%, sodium cholate 0.2%, calcium bicarbonate 0.6, stone powder 0.4%.

[0097] 2.2 Establishment of type 2 diabetes rat model: on the 29th day of gavage, rats in each group except the blank group were fasted but not watered for 24 hours, and then intraperitoneally injected with pyrimidines at a dose of 107mg·kg -1 , and then continued to be fed with high-energy feed for 5 days.

[0098] 2.3 Detection index

[0099] 2.3.1 Determination of fasting blood glucose of experimental rats: on the 28th day (before modeling) and the 32nd day (after modeling) of gavage, all experimental rats were fasted for 4 hours, and the tail vein blood was taken to measure the fasting blood glucose value. The results are shown in Table 4.

[0100] 2.3.2 Determination of liver glycogen After the rats were sacrificed, the liver was separated by opening the abdominal cavity, a small piece of liver tissue was taken, and the liver glycogen content was measured according to the instruction manual of the liver glycogen determination kit (product A043-1-1, Nanjing Jiancheng Biological Engineering Research Institute Co., Ltd.). The results are shown in Table 5 and Figure 3 .

[0101] Table 4

[0102]

[0103] From Table 4, before modeling (28 days), the fasting blood glucose of the blank group, the model group, the positive control group and each treatment group fluctuated within the normal range, and there was no significant difference between groups; after modeling (32 days), the fasting blood glucose of the model group, the positive control group and each treatment group showed an upward trend, compared with before modeling (28 days), the blood glucose of the model group increased the most, and the average fasting blood glucose was as high as 16.76 mmol / L, and the blood glucose of the model group was significantly higher than that of the blank group, indicating that the modeling of diabetic rats was successful. After modeling, the fasting blood glucose of each treatment group was lower than that of the model group; compared with the collagen peptide fermentation liquor and the plant enzyme group, the fasting blood glucose of Example 2 was significantly reduced, indicating that the synergistic effect of the collagen peptide fermentation liquor and the plant enzyme could significantly reduce blood glucose, and the effect of combined use was more obvious than that of single use; compared with Comparative Examples 1-7, the fasting blood glucose of Example 2 was significantly reduced, indicating that the collagen peptide fermentation liquor and the plant enzyme together could reduce blood glucose, and only the collagen peptide fermentation liquor fermented by Lactobacillus rhamnosus NCU2217 and the plant enzyme synergistically fermented by Ganoderma lucidum, Alisma orientale and Radix Menyanthis could significantly reduce blood glucose. Therefore, the collagen peptide composition obtained by the application has the effect of reducing blood glucose by synergistic effect of collagen peptide fermentation liquor and plant enzyme.

[0104] Table 5

[0105]

[0106] From Table 5 and Figure 3 Compared with the blank group, the liver glycogen content of the model group was significantly reduced, indicating that the model group of rats had glucose metabolism disorder. Compared with the model group, the liver glycogen content of the positive control group and each treatment group was significantly increased, and the positive control group and each treatment group could regulate the balance of glucose metabolism. Compared with the collagen peptide fermentation liquor and the plant enzyme group, the liver glycogen content of Example 2 was significantly increased, indicating that the synergistic effect of the collagen peptide fermentation liquor and the plant enzyme could significantly regulate the balance of glucose metabolism, and the effect of combined use was more obvious than that of single use. Compared with Comparative Examples 1-7, the liver glycogen content of Example 2 was significantly increased, indicating that the collagen peptide fermentation liquor and the plant enzyme together could significantly regulate the balance of glucose metabolism, and only the collagen peptide fermentation liquor fermented by Lactobacillus rhamnosus NCU2217 and the plant enzyme synergistically fermented by Ganoderma lucidum, Alisma orientale and Radix Menyanthis could significantly reduce the effect. Therefore, continuous intragastric administration of the collagen peptide composition obtained by the application improves the ability of rats to handle high blood glucose, which helps to regulate glucose metabolism.

[0107] III. Protective effect of collagen peptide composition on acute liver injury of mice

[0108] 3.1 Experimental grouping and administration: Eight-week-old male mice C57BL / 6 were purchased from Changzhou Cavens Experimental Animal Co., Ltd.; 78 mice were randomly divided into 13 groups, namely 1 blank group, 1 model group, 1 positive control group (bifendate), 10 treatment groups (Example 2, Comparative Examples 1-7 groups, collagen peptide fermentation liquor and plant enzyme). The blank group and the model group were given physiological saline by gavage, and the gavage dose was: physiological saline 10 mL·kg -1 (bw / d); the positive control group was given bifendate drop pills by gavage at a dose of: 200 mg·kg -1 (bw / d), and was given by gavage at a dose of 10 mL·kg -1 (bw / d); the treatment groups were given the compositions obtained in Example 2 and Comparative Examples 1-7 by gavage at a dose of 10 mL·kg -1 (bw / d) after being diluted 10 times with distilled water. The collagen peptide fermentation liquor and the plant enzyme obtained in Example 2 were diluted according to the amount in Example 2 (the collagen peptide fermentation liquor was diluted 10*12 / 15 times, and the plant enzyme was diluted 10*3 / 15 times), and were given by gavage at a dose of 10 mL·kg -1 (bw / d). Bifendate drop pills were provided by Beijing Union Pharmaceutical Factory.

[0109] Except that the normal group and the model group were given physiological saline by gavage, the rest of the groups were given drugs by gavage for 10 days, 1 hour after the last administration, the normal group was injected intraperitoneally with olive oil, and the other groups of mice were injected intraperitoneally with 0.3% CCl4 olive oil solution 10 ml / kg, and were fasted but not watered. After 24 hours, the weight was measured, the blood was taken from the eye orbit after anesthesia, and the mice were sacrificed after dissection to take the liver.

[0110] 3.2 Detection index

[0111] 3.2.1 Before the mice were sacrificed, the weight of all mice in each model group was measured, all mice were sacrificed after anesthesia and blood collection, and the liver was taken out, washed with 4℃ physiological saline, absorbed with filter paper, and the liver weight was measured. After collecting the data, the liver coefficient of each group of mice was calculated, and the results are shown in Table 6.

[0112] Liver coefficient = liver weight (g) / body weight (g) × 100%

[0113] 3.2.2 Biochemical index

[0114] The mouse blood obtained by the eye orbit blood collection method was placed in a 4 mL coagulation tube, centrifuged at 25℃, 3500 rpm for 15 min, and the supernatant was taken to obtain serum. The mouse serum ALT and AST were measured using the corresponding kit according to the instruction method, and the results are shown in Table 6 and Figure 4The same part of a small piece of liver tissue was taken, and pre-cooled normal saline was added at 1:9. After homogenization, the supernatant obtained by centrifugation at 4°C and 9500 rpm for 20 min was 10% liver tissue homogenate, and the SOD and MDA contents in the liver tissue were determined according to the corresponding kit operation instructions, and the results are shown in Table 7 and Figure 5 The alanine aminotransferase (ALT) test kit, aspartate aminotransferase (AST) test kit, malondialdehyde (MDA) kit, and superoxide dismutase (SOD) kit were all purchased from Shanghai Blue Sky Bio-technology Co., Ltd.

[0115] Table 6

[0116]

[0117] Table 7

[0118]

[0119] It can be seen from Table 6 and Figure 4 Compared with the blank group, the liver index of the model group mice was significantly increased, and the serum ALT and AST contents of the model group mice were significantly increased, indicating that the liver of the model group mice was damaged. Compared with the model group, the liver index, ALT and AST contents of each treatment group were decreased, and the liver index, serum ALT and AST contents of the positive control group and Example 2 group mice were significantly decreased. Compared with the collagen peptide fermentation liquor and plant enzyme group, the liver index, serum ALT and AST contents of the mice in Example 2 group were significantly decreased, indicating that the synergistic effect of collagen peptide fermentation liquor and plant enzyme could significantly reduce the liver index, serum ALT and AST contents of mice, and the effect of combined use was more obvious than that of single use; compared with Comparative Examples 1-7, the liver index, serum ALT and AST contents of mice in Example 2 group were significantly decreased, indicating that collagen peptide fermentation liquor and plant enzyme together could significantly reduce the liver index, serum ALT and AST contents of mice, and only the collagen peptide fermentation liquor fermented by Lactobacillus rhamnosus NCU2217 and the plant enzyme synergistically fermented by Ganoderma lucidum, Alisma orientale and Radix Solimarii could have a more significant synergistic effect of collagen peptide fermentation liquor and plant enzyme.

[0120] It can be seen from Table 7 and Figure 5It can be seen that compared with the blank group, the SOD activity in the liver of the model group mice was significantly reduced, indicating that the liver of all model group mice was damaged by oxidation; the MDA content in the liver of the model group mice was significantly increased, indicating that all model group mice had severe lipid peroxidation in the liver. Compared with the model group, the SOD activity of each treatment group was restored, and the MDA content was decreased, and the SOD activity of the positive control group and the liver of the mice in Example 2 was significantly increased, and the MDA content was significantly decreased. Compared with the collagen peptide fermentation liquor and plant enzyme group, the SOD activity of the liver of the mice in Example 2 was significantly increased, and the MDA content was significantly decreased, indicating that the synergistic effect of collagen peptide fermentation liquor and plant enzyme could significantly increase the SOD activity and reduce the MDA content in the liver of the mice, and the effect of combined use was more obvious than that of single use; compared with Comparative Examples 1-7, the SOD activity of the liver of the mice in Example 2 was significantly increased, and the MDA content was significantly decreased, indicating that the collagen peptide fermentation liquor and the plant enzyme together could significantly increase the SOD activity and reduce the MDA content in the liver of the mice, and only the collagen peptide fermentation liquor fermented by Lactobacillus rhamnosus NCU2217 and the plant enzyme synergistically fermented by Ganoderma lucidum, Alisma orientale and Radix Alcoholyciae were used, and the synergistic effect of collagen peptide fermentation liquor and plant enzyme was more significant.

[0121] In summary, the collagen peptide composition obtained by the present application can significantly alleviate the liver enlargement of acute liver injury model mice, reduce the content of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in the serum of the model mice, and relieve the degree of liver damage. The activity of antioxidant enzyme system was determined, and it was found that the collagen peptide composition obtained by the present application can significantly improve the level of SOD in the liver of acute liver injury model mice and reduce the content of MDA, thereby relieving the oxidative stress of the liver of the model mice.

[0122] The above-described embodiments only express the specific implementation of the present application, which is described in more detail and in more detail, but it cannot be understood as limiting the scope of the patent of the present application. It should be noted that for ordinary skilled in the art, without departing from the concept of the present application, a number of modifications and improvements can be made, which are within the scope of protection of the present application.

Claims

1. A collagen peptide composition for regulating metabolism and promoting weight loss and fat reduction, characterized in that, The collagen peptide fermentation liquor and the plant enzyme have a volume ratio of 10-15:2-5; the collagen peptide fermentation liquor is obtained by fermenting collagen peptide with Lactobacillus rhamnosus; the plant enzyme is obtained by fermenting a traditional Chinese medicine composition with Lactobacillus casei; the specific fermentation method of the collagen peptide fermentation liquor is as follows: 3-5% (w / v) of collagen peptide is added to water, and Lactobacillus rhamnosus is inoculated, and then fermentation is carried out at 30-42 ℃ for 16-24 h to obtain the collagen peptide fermentation liquor; the Lactobacillus rhamnosus is Lactobacillus rhamnosus NCU2217, which is preserved in the China General Microbiological Culture Collection Center with a preservation number of CGMCC NO 15573; the traditional Chinese medicine composition is composed of Ganoderma lucidum, Alisma orientale and Radix Gastrodiae; the weight ratio of Ganoderma lucidum, Alisma orientale and Radix Gastrodiae is 3:2:2; The specific preparation method of the plant enzyme is as follows: (a) the traditional Chinese medicine materials in the traditional Chinese medicine composition are mixed according to the proportion, and then added to distilled water for extraction; the obtained extract is concentrated to obtain a traditional Chinese medicine concentrate; (b) Lactobacillus casei is directly inoculated into the traditional Chinese medicine concentrate for inoculation fermentation to obtain the plant enzyme.

2. The collagen peptide composition for regulating metabolism and reducing weight and fat according to claim 1, characterized in that, The volume ratio of the collagen peptide fermentation liquor and the plant enzyme is 12:

3. 3.The collagen peptide composition for regulating metabolism and reducing weight and fat according to claim 1, characterized in that, The inoculation amount of the Lactobacillus rhamnosus is 1 x 10 6 ~1 x 10 7 cfu / mL. 4.The collagen peptide composition for regulating metabolism and reducing weight and fat according to claim 1, characterized in that, The inoculation amount of the Lactobacillus casei is 1 x 10 6 1 x 10 7 cfu / mL.

5. Use of the collagen peptide composition of claim 1 in the manufacture of a medicament, characterized in that, The medicine is a medicine with the effects of regulating metabolism and reducing weight and fat.

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