CN114631628A - Method for increasing concentration of FGF21 in blood - Google Patents

Abstract

The invention provides a method for increasing the concentration of FGF21 in blood, which comprises n cycles, wherein each cycle has at least one low protein intake period, and n is a positive integer greater than or equal to 1. The method of the present invention is effective in improving glycolipid metabolism in the body by increasing the level of FGF21 in vivo, and can be used for treating or preventing metabolic diseases.

Description

A method for increasing blood FGF21 concentration

technical field

The present invention belongs to the field of nutrition, and more particularly, the present invention relates to a method for increasing blood FGF21 concentration.

Background technique

Fibroblast growth factor 21 (FGF21) is a new member of the FGF family, which is closely related to the regulation of glucose and lipid metabolism. The original study found that the mRNA of FGF21 was mainly expressed in the liver of mice. Later, some scholars found that FGF21 is also expressed in muscle and fat. In addition, FGF21 is also present in beta cells of the pancreas, and at higher levels than in liver and fat.

Recent studies have found that FGF21 can improve the body's glucose and lipid metabolism, and can be used to intervene in metabolic diseases including diabetes, prediabetes, fatty liver, dyslipidemia, etc.

At present, many drug companies are conducting research and development of FGF21, mainly synthesizing FGF21 recombinant protein, or synthesizing FGF21 recombinant protein with longer half-life, and injecting it into the body to achieve the purpose of improving the function of glucose and lipid metabolism. The main disadvantage of these methods is the need for long-term injection of recombinant protein, which is expensive and may have side effects.

Therefore, there is an urgent need in the art to develop an effective, side-effect-free and low-cost method for increasing FGF21 levels in vivo.

SUMMARY OF THE INVENTION

The purpose of the present invention is to provide an effective, non-side effect, and low-cost method for increasing the level of FGF21 in vivo.

In a first aspect of the present invention, there is provided a low protein diet therapy or method of increasing the level (eg concentration) of FGF21 in vivo, said therapy or method comprising n cycles, each cycle at least one low protein intake period, wherein n is a positive integer ≥1.

In another preferred example, n is a positive integer greater than or equal to 2, preferably, n is 2-12, preferably, n is 3-8, more preferably, n is 3-6.

In another preferred embodiment, each cycle further includes at least one high protein intake period.

In another preferred embodiment, each cycle further includes at least one normal protein intake period.

In another preferred embodiment, the low protein intake period (t _low ) is greater than or equal to 1 day, preferably, 2-7 days, more preferably, 2-5 days, more preferably, 2-3 days.

In another preferred embodiment, the high protein intake period (t _high ) is greater than or equal to 1 day, preferably, 2-7 days, more preferably, 2-5 days, more preferably, 3-4 days.

In another preferred embodiment, the normal protein intake period (t _normal ) is greater than or equal to 1 day, preferably, 2-7 days, more preferably, 2-5 days, more preferably, 4-5 days.

In another preferred example, t _low + t _normal ≤ 10 days, preferably, t _low + t _normal is 4-9 days, more preferably, 5-8 days.

In another preferred embodiment, t _low + t _high ≤ 10 days, preferably, t _low + t _high is 4-9 days, more preferably, 5-8 days.

In another preferred embodiment, the therapy or method is aimed at improving the function of FGF21 on glucose and lipid metabolism in the body, for preventing and/or treating metabolic diseases.

In another preferred embodiment, the metabolic disease is selected from the group consisting of diabetes, prediabetes, fatty liver, dyslipidemia, or a combination thereof

In another preferred example, the concentration of low protein intake during the low protein intake period is <7.5%, preferably 1-5%, more preferably 2-5%.

In another preferred embodiment, the high protein refers to I1/I0 ≥ 1.5, preferably ≥ 2, more preferably ≥ 3, wherein I1 is the high protein intake of the individual in one day, and I0 is the normal daily intake of the individual. Protein intake (I0).

In another preferred embodiment, the high protein means that the total protein intake of mammals (eg, humans) in one day is higher than 90 g, preferably higher than 100 g.

In another preferred embodiment, the low protein refers to I2/I0≤50%, preferably≤30%, more preferably,≤25%, more preferably,≤20%, wherein I2 is the low protein of the individual for one day Intake, I0 is the normal protein intake (I0) of the individual in one day.

In another preferred embodiment, the low protein means that the total protein intake of mammals (such as humans) in one day is less than 30 g, preferably less than 25 g.

In another preferred embodiment, each cycle includes at least one low protein intake period.

In another preferred embodiment, each cycle includes at least one low protein intake period and at least one high protein intake period.

In another preferred embodiment, each cycle includes at least one low protein intake period and at least one normal protein intake period.

In another preferred embodiment, the therapy includes therapy of metabolic diseases.

In another preferred embodiment, the therapy includes increasing the level of FGF21 in the blood and improving the metabolism of glucose and lipids in the body, so as to prevent and treat metabolic diseases.

In another preferred embodiment, the metabolic disease is selected from the group consisting of diabetes, prediabetes, fatty liver, dyslipidemia, or a combination thereof.

In another preferred embodiment, the therapy or method is non-therapeutic and non-diagnostic.

The second aspect of the present invention provides the use of a low-protein diet for preparing a composition or a medicine for increasing the level of FGF21 in the body and improving the metabolism of glucose and lipids in the body through low-protein food therapy, thereby treating Metabolic disease.

In another preferred embodiment, the composition or medicine is also used for one or more purposes selected from the following group:

(a) increase the level of FGF21 in the blood;

(b) increase insulin sensitivity;

(c) Improve fatty liver;

(d) increase the basal metabolic rate;

(e) Improve dyslipidemia;

(f) Weight loss.

In another preferred example, the composition includes a food composition and a pharmaceutical composition.

In another preferred embodiment, the composition includes dietary supplements.

In another preferred example, the composition further includes a high-protein meal.

In another preferred embodiment, the composition further includes a normal protein meal.

In another preferred example, the composition includes a low-protein meal and a high-protein meal.

In another preferred embodiment, the composition includes a low-protein meal and a normal protein meal.

In another preferred embodiment, the composition includes n groups of compositions, wherein n≥1 (preferably, n is 2-12, preferably, n is 3-8, more preferably, n is 3- 6), and each group included a low-protein diet.

In another preferred embodiment, the low protein concentration in the low protein meal is <7.5%, preferably 1-5%, more preferably 2-5%.

In another preferred embodiment, the high-protein diet refers to I1'/I0'≥1.5, preferably ≥2, more preferably ≥3, wherein I1' is the protein content in the high-protein diet, and I0' is The protein content of a normal diet.

In another preferred example, the high-protein diet refers to the total protein intake of mammals (such as humans) in one day is higher than 90g (preferably 90g-120g), preferably higher than 100g (preferably 100g-150g) .

In another preferred embodiment, the low-protein diet refers to I2'/I0'≤50%, preferably≤30%, more preferably,≤25%, more preferably,≤20%, wherein I2' is low The protein content in the protein diet, I0' is the protein content in the normal diet.

In another preferred embodiment, the low-protein diet refers to that the total protein intake of mammals (such as humans) in one day is less than 30g (preferably 25-30g), preferably less than 25g (preferably 20-25g) .

The third aspect of the present invention provides the use of a product for preparing a composition or a medicament for increasing the level of FGF21 in the body through low-protein food therapy, improving the metabolism of glucose and lipids in the body, thereby treating metabolic disorders disease, wherein the product comprises a low protein product.

In another preferred embodiment, the product also includes a high-protein product.

In another preferred embodiment, the product further includes a normal protein product.

In another preferred embodiment, the products include low-protein products and high-protein products.

In another preferred embodiment, the products include low protein products and normal protein products.

In another preferred embodiment, the product includes n groups of compositions, wherein n≥1 (preferably, n is 2-12, preferably, n is 3-8, more preferably, n is 3-6 ), and each group included low protein products.

In another preferred embodiment, the groups also include high-protein products.

In another preferred embodiment, the groups also include normal protein products.

In another preferred embodiment, the high-protein product refers to I1"/I0"≥1.5, preferably ≥2, more preferably ≥3, wherein I1" is the protein content in the high-protein product, and I0" is Protein content in normal products.

In another preferred embodiment, the high protein product refers to the protein content in the product >90g (preferably 90g-120g), preferably >100g (preferably 100g-150g).

In another preferred embodiment, the low protein product refers to I2"/I0"≤50%, preferably≤30%, more preferably,≤25%, more preferably,≤20%, wherein I2" is low The protein content in the protein product, I0" is the protein content in the normal product.

In another preferred embodiment, the low-protein product refers to the protein content in the product <30g (preferably 25-30g), preferably <25g (preferably 20-25g).

In another preferred embodiment, the low protein concentration in the low protein product is <7.5%, preferably 1-5%, more preferably 2-5%.

A fourth aspect of the present invention provides a product, comprising n groups of products, wherein n≥1 (preferably, n is 2-12, preferably, n is 3-8, more preferably, n is 3-6 ), and each group included low protein products.

In another preferred embodiment, the groups also include high-protein products.

In another preferred embodiment, the groups also include normal protein products.

In another preferred embodiment, the groups include low-protein products and high-protein products.

In another preferred embodiment, the groups include low protein products and normal protein products.

In another preferred embodiment, the product includes dietary supplements.

In another preferred embodiment, the low protein concentration in the low protein product is <7.5%, preferably 1-5%, more preferably 2-5%.

It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described in the following (eg, the embodiments) can be combined with each other to constitute new or preferred technical solutions. Due to space limitations, it is not repeated here.

Description of drawings

Figure 1 shows that an intermittent low-protein diet can significantly increase blood FGF21 levels in mice with type 1 diabetes.

Among them, mice were grouped: STZ-treated mice were divided into 7 groups. The first group ate ordinary food freely every day; the other 6 groups of mice ate freely mouse food with different protein concentrations for the first 3 days of the week, and the latter 4 days for ordinary food freely. food.

Figure 2 shows that intermittent low protein (5%) diet can significantly increase FGF21 levels in the blood of type 2 diabetic mice.

Among them, WT: mice ate ordinary food ad libitum every day; CTRL: db/db mice ate ordinary food ad libitum every day; HPD: mice ate low-protein food for the first three days of the week, and high-protein food for the last 4 days; NC: Mice were fed a low-protein diet for the first 3 days of the week and a normal diet for the last 4 days.

Figure 3 shows that an intermittent low-protein (5%) diet can significantly improve muscle insulin sensitivity in type 2 diabetic mice.

Among them, 1: db/db mice ate ordinary food ad libitum every day; 2: db/db mice ate low-protein food for the first three days of the week, and high-protein food for the last 4 days; 3: db/db mice every day Eat low-protein food for the first 3 days of the week and regular food for the next 4 days.

Detailed ways

After extensive and in-depth research, the inventors have unexpectedly discovered for the first time that intermittent use of a dietary regimen containing low-protein foods can effectively improve insulin sensitivity by increasing the level of FGF21 in the body (such as blood), thereby effectively treating metabolic disorders. disease, and the method of the present invention has no side effects and low cost. On this basis, the present inventors have completed the present invention.

low protein diet

In the present invention, low-protein food therapy refers to using low-protein diet in one cycle to improve the body's glucose and lipid metabolism by increasing the level of FGF21 in the body (eg, blood).

In the present invention, the low protein diet may include at least one low protein intake period.

In a preferred embodiment, the low protein food regimen includes at least one low protein intake period, at least one high protein intake period.

In a preferred embodiment, the low protein food regimen includes at least one low protein intake period, at least one normal protein intake period.

low protein meal

In the present invention, low-protein diet refers to more than 50% of the normal recommended amount of diet (the daily recommended amount of protein for human body is 60g, then the daily intake needs to be lower than 30g, preferably lower than 25g).

high protein meal

In the present invention, high protein diet refers to more than 50% of the normal recommended amount of the diet (the daily recommended amount of protein for a human body is 60 g, and the daily intake needs to exceed 90 g).

normal protein meal

In the present invention, normal protein diet refers to the recommended amount of normal diet (the daily recommended amount of protein for human body is 60 g).

Dietary Supplements

The present invention also provides a dietary supplement for alleviating or treating various metabolic diseases, which contains a low-protein product.

The main advantages of the present invention include:

(1) The present invention finds for the first time that the low-protein food therapy of the present invention can effectively treat metabolic diseases by increasing the level of FGF21 in the body (eg, blood), and the method of the present invention has no side effects and low cost.

(2) The present invention finds for the first time that the low-protein food therapy of the present invention can improve insulin sensitivity.

(3) The present invention finds for the first time that for the STZ-induced type 1 diabetes mouse model, intermittent use of a diet with a protein concentration lower than 7.5% can significantly increase the concentration of FGF21 in the blood.

(4) The present invention finds for the first time that, for the mouse model of type 2 diabetes, intermittent use of a low-protein diet can significantly increase the concentration of FGF21 in the blood.

(5) The present invention finds for the first time that, in a mouse model of type 2 diabetes, intermittent use of a low-protein diet increases the insulin sensitivity of mouse skeletal muscle.

(6) The present invention provides a non-drug dietary method for increasing FGF21 in the body, which can be used to improve glucose and lipid metabolism in the body, and to interfere with metabolic diseases including diabetes, prediabetes, fatty liver, dyslipidemia, and the like.

The present invention will be further described below in conjunction with specific embodiments. It should be understood that these examples are only used to illustrate the present invention and not to limit the scope of the present invention. The experimental method of unreceipted specific conditions in the following examples, usually according to routine conditions such as J. Sambrook and other editors, Molecular Cloning Experiment Guide, Science Press, described in 2002 (New York:Cold Spring Harbor Laboratory Press, 2002) conditions, or as recommended by the manufacturer. Percentages and parts are weight percentages and parts unless otherwise specified.

The materials and reagents used in the examples are all commercially available products unless otherwise specified.

general method

Materials and Methods:

1.1 Materials: 8-week-old male C57BL/6 mice and db/db mice were purchased from Slack Laboratory Animal Co., Ltd.; STZ was purchased from Sigma Company; FGF21 detection kit was purchased from R&D Company.

1.2 Methods

1.2.1 STZ to build a type 1 diabetes model: C57 mice fasted for 5 hours were injected with STZ (40 mg/kg body weight) for 5 consecutive days. After one week, the blood glucose of the mice was observed to rise significantly, and the model was successfully established;

1.2.2 db/db mice to construct a type 2 diabetes model: db/db mice purchased from the company at the age of 5-6 weeks were fed with common chow in the mouse room for about 10 weeks to detect the blood sugar level. When the average blood sugar level was higher than 11.1 mmol/l, nutritional intervention was started.

1.2.3 Experimental scheme:

Experiment 1: 15 8-week-old male C57BL/6 mice, 5 in each group, were divided into three groups. The mice in the first group ate ordinary food ad libitum every day; the mice in the second group ate low-protein (5%) food for the first three days of the week and normal food for the last 4 days; the mice in the third group were fed with medium protein for the first three days of the week Food content (20%), followed by regular food for the next 4 days; repeated weekly for a total of 5 weeks.

Experiment 2: A total of 18 10-week-old male db/db mice, 6 in each group, and 6 WT control mice, were divided into four groups. The first group of WT mice ate ordinary food ad libitum every day; the second to fourth groups were db/db mice. Mice in the CTRL group also ate ordinary food ad libitum every day. The third group of mice ate low-protein food for the first three days of the week and high-protein food for the last 4 days; the fourth group of mice ate low-protein food for the first 3 days of the week and normal food for the last 4 days; repeated every week for a total of Lasts 4 weeks.

1.3 Rat food formula

1.3.1 Rat diets with different protein concentrations (for the type 1 diabetes mouse model) and 40% high protein feed were purchased from Shanghai Fanbo Biotechnology Co., Ltd. The main nutrients are casein, corn starch, sucrose, maltodextrin, cellulose, soybean oil, vitamin and mineral mixture, etc.

1.3.2 Low-protein mouse food formula (for type 2 diabetes mouse model): self-designed, as follows:

Test results: protein 5%, fat 12.3%, total sugar 26.7%.

1.3.3 Normal rat food formula: 20.5% normal rat food was purchased from Shanghai Proton Biotechnology Co., Ltd. The main nutrients are fish meal, wheat, corn, bran, vitamins, minerals, amino acids, etc.

Example 1 Intermittent low-protein diet can significantly increase the level of FGF21 in the blood of type 1 diabetic mice

The 8-week-old C57 male mice were first constructed to build a type 1 diabetes model: STZ injected (40 mg/kg body weight) after fasting for 5 hours was administered for 5 consecutive days. After a week, the blood sugar of the mice was observed to rise significantly, and the model was successfully established; the control group was small. Rats were given free access to normal food every day; other groups were given free access to mouse food with different protein concentrations (including 0%, 2.5%, 5%, 7.5%, 10%, and 20%) on the first 3 days of the week, and normal rats on the last 4 days. grain. After 5 consecutive weeks of dietary intervention, we sacrificed the STZ-modeled type 1 diabetic mice, collected blood, separated serum, and detected the changes in serum FGF21 level with ELISA kit. Serum FGF21 levels were found to be significantly elevated in mice fed intermittently with 0%, 2.5% and 5% low protein concentration rat chow after 5 weeks of dietary intervention (Figure 1). Therefore, this result suggests that intermittent use of a diet below 7.5% protein concentration can significantly increase blood FGF21 levels in mice.

Example 2 Intermittent low-protein (5%) diet can significantly increase blood FGF21 levels in type 2 diabetic mice

Next, we tested this finding in another diabetes model. We used db/db mice with type 2 diabetes. The 5-6 weeks old db/db mice purchased from the company were fed with ordinary chow in the mouse room until about 10 weeks to detect the blood sugar level. When the average blood sugar level was higher than 11.1mmol/l, nutritional intervention was started. The C57 mice in the WT group ate ordinary food ad libitum every day; the CTRL group were db/db mice, and the mice in the CTRL group also ate ordinary food ad libitum every day; the mice in the HPD group were db/db mice, and ate low protein for the first three days of the week The mice in the NC group were also db/db mice, and were fed with low-protein food for the first 3 days of the week and normal food for the last 4 days. The mice were sacrificed after 4 consecutive weeks of dietary intervention, and the blood was collected to separate the serum to detect the changes of FGF21 levels in serum by ELISA kit. We found that after 4 weeks of dietary intervention, the serum levels of FGF21 in mice in the intermittent low-protein group were significantly higher than those in the WT and CTRL control groups (Figure 2). Therefore, this result revealed that intermittent low-protein diet can significantly increase the level of FGF21 in the blood of mice.

Example 3 Increased Skeletal Muscle Insulin Sensitivity Using an Intermittent Low-Protein Diet in Type 2 Diabetic Mice

Increased levels of FGF21 are known to improve insulin sensitivity in peripheral tissues. We then examined the insulin sensitivity of skeletal muscle in db/db mice after nutritional intervention. The 5-6 weeks old db/db mice purchased from the company were fed with ordinary chow in the mouse room until about 10 weeks to detect the blood sugar level. When the average blood sugar level was higher than 11.1mmol/l, nutritional intervention was started. The first group (group 1) was the control group of mice, and they ate ordinary food ad libitum every day; the second group of mice (group 2) ate low-protein food for the first 3 days of the week and high-protein food for the last 4 days; Rats (group 3) were fed with low-protein food for the first 3 days of the week, and normal food for the last 4 days. Mice were sacrificed after 4 consecutive weeks of dietary intervention. Before sacrifice, mice were injected intraperitoneally with 4 U of insulin to stimulate insulin signaling. After that, protein was extracted from mouse skeletal muscle, and Western Blot experiment was performed to detect the content of phosphorylated and total AKT protein in muscle tissue. It was found that after 4 weeks of intermittent low-protein dietary intervention, the content of phosphorylated AKT in muscle tissue of mice was significantly up-regulated (Figure 3). Since ATK is one of the most critical proteins in the insulin signaling pathway, the results suggested that an intermittent low-protein diet could improve insulin sensitivity in mice.

All documents mentioned herein are incorporated by reference in this application as if each document were individually incorporated by reference. In addition, it should be understood that after reading the above teaching content of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of the present application.

Claims (10)

What is claimed is: 1. A low-protein food therapy or a method for increasing the level of FGF21 in vivo, wherein the therapy or method comprises n cycles, each cycle having at least one low-protein intake period, wherein n is a positive integer ≥ 1. 2. The therapy or method according to claim 1, wherein n is a positive integer ≥ 2, preferably, n is 2-12, preferably, n is 3-8, more preferably, n 3-6. 3. The therapy or method of claim 1, wherein each cycle further comprises at least one period of high protein intake. 4. The therapy or method of claim 1, wherein each cycle further comprises at least one period of normal protein intake. 5. The therapy or method according to claim 1, wherein the low protein intake period (t _low ) is greater than or equal to 1 day, preferably 2-7 days, more preferably 2-5 days, More preferably, 2-3 days. 6. The therapy of claim 3, wherein the high protein intake period (t _high ) is greater than or equal to 1 day, preferably, 2-7 days, more preferably, 2-5 days, more preferably ground, 3-4 days. 7. The therapy or method according to claim 1, wherein the concentration of low protein ingested during the low protein intake period is <7.5%, preferably 1-5%, more preferably 2 -5%. 8. The purposes of a low-protein meal, characterized in that, for the preparation of a composition or a drug, the composition or drug is used to improve the level of FGF21 in vivo by low-protein food therapy, improve the function of glucose and lipid metabolism in the body, and thereby treat metabolism sexually transmitted diseases. 9. Use of a product, characterized in that, for the preparation of a composition or a medicament, the composition or medicament is used to increase the level of FGF21 in vivo by low-protein food therapy, improve the metabolism of glucose and lipids in the body, and thereby treat metabolic diseases , wherein the product includes a low-protein product. 10. A product, characterized by comprising n groups of products, wherein n≥1 (preferably, n is 2-12, preferably, n is 3-8, more preferably, n is 3-6) , and each group included low-protein products.

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