CN118203566A - Application of 2-aminoadipic acid in regulating inflammatory response induced by high-fat diet - Google Patents

Abstract

The invention discloses application of 2-amino adipic acid in regulating inflammatory response induced by high-fat diet, solves the problem of chronic inflammation of organisms caused by life style and dietary habit changes at present, and discovers in researches that the 2-amino adipic acid can obviously inhibit expression and secretion of tumor necrosis factor-alpha, promote expression and secretion of transforming growth factor-beta, effectively reduce activation level of nuclear factor kappa B signal channel and finally achieve the technical effect of improving chronic inflammatory state induced by high-fat diet. The invention provides a powerful research platform for treating and preventing inflammatory reaction and predicts that the invention has wide prospect in clinical application.

Description

Application of 2-aminoadipic acid in regulating inflammatory response induced by high-fat diet

Technical Field

The present invention relates to the field of biomedical technology, and in particular to application of 2-aminoadipic acid in regulating inflammatory response induced by a high-fat diet.

Background technique

Chronic inflammation is a series of pathological reactions produced by the body under the influence of persistent internal and external inflammatory factors or local injury stimulation. This pathological state may be a sign of acute disease onset, may also constitute the basis of certain chronic diseases, and may promote the occurrence, malignant evolution, invasion and metastasis of cancer. Empirical studies have shown that chronic inflammation is closely related to the development of many diseases, including atherosclerosis, type 2 diabetes, rheumatoid arthritis, systemic lupus erythematosus, multiple sclerosis, Parkinson's disease, Crohn's disease, colon cancer and liver cancer. Chronic inflammation not only contributes to the development of the disease, but may also aggravate the condition of existing diseases. Therefore, controlling chronic inflammation has become a key strategy for preventing and treating these diseases.

Inflammatory factors refer to a class of protein molecules that trigger and regulate inflammatory responses when the body's tissues are damaged or infected, mainly including cytokines, chemokines and other regulatory factors. These factors promote the progression of inflammation during cell-to-cell interactions, cell migration and cell proliferation, and they play a vital role in protecting the body from infection and injury. However, excessive or long-term inflammatory responses may lead to a variety of diseases. In the upstream of the inflammatory cascade, tumor necrosis factor-α (TNF-α) and transforming growth factor-β (TGF-β) play a core role; treatment against them has achieved clinical results in diseases such as arthritis, psoriasis, and inflammatory bowel disease. Therefore, in order to solve the current lack of products in the inflammatory response induced by a high-fat diet, an effective and promising food, medicine or health product is urgently needed to meet the needs of scientific research and the market.

2-Aminoadipic acid (2-AAA) is selectively toxic to glial cells and can specifically inhibit the activation of astrocytes. Further studies have shown that 2-AAA plays a role in the regulation of glucose homeostasis and lipid metabolism, and is associated with a variety of pathological processes such as atherosclerosis. It can be seen that regulating 2-AAA levels to intervene in chronic inflammation caused by a high-fat diet can provide new research directions and treatment methods for disease treatment.

Summary of the invention

In view of the problems existing in the prior art, the present invention provides the use of 2-aminoadipic acid in regulating the inflammatory response induced by a high-fat diet, and achieves the purpose of regulating the body's inflammatory response by studying the effect of 2-aminoadipic acid on TNF-α, TGF-β levels and NF-κB signaling pathway activity.

To achieve the above object, the present invention provides the following technical solutions:

The present invention has found in the research that 2-AAA can inhibit the production of TNF-α and promote the production of TGF-β, and inhibit the activation of the NF-κB signaling pathway caused by oxidized low-density lipoprotein (ox-LDL), inhibit the inflammatory response, and finally achieve the technical effect of improving the chronic inflammation of the body induced by a high-fat diet. Excessive and abnormal activation of the NF-κB signaling pathway is an important process for the occurrence of inflammatory response, so the reduction of NF-κB pathway activity can be regarded as an important sign of the inhibition of inflammatory response.

The present application provides the use of 2-aminoadipic acid in regulating the inflammatory response induced by a high-fat diet, wherein the 2-aminoadipic acid can reduce the level of TNF-α and increase the level of TGF-β in serum.

Furthermore, the 2-aminoadipic acid can inhibit the activation of the NF-κB signaling pathway induced by ox-LDL, thereby inhibiting the chronic inflammatory response.

Furthermore, the 2-aminoadipic acid can improve the inflammatory response of macrophages caused by high fat.

Furthermore, the 2-aminoadipic acid can improve chronic inflammation caused by lipid metabolism disorders.

The present invention also protects a medicine, food or health product for regulating inflammation of the body, comprising 2-aminoadipic acid and/or its derivatives.

Furthermore, the food, medicine or health product comprises an effective amount of 2-aminoadipic acid and/or its derivatives and acceptable excipients.

Beneficial effects achieved by the present invention:

The application of 2-aminoadipic acid provided by the present invention in regulating the inflammatory response induced by a high-fat diet, aims at the current situation of frequent chronic inflammation of the body caused by unhealthy lifestyles and eating habits, and provides a new product for inhibiting inflammation of the body by inhibiting the activation of the NF-κB pathway. 2-AAA can inhibit the activity of the NF-κB signaling pathway, regulate the levels of inflammatory factors TNF-α and TGF-β in the body, thereby inhibiting the inflammatory response of the body, and finally achieving the technical effect of improving chronic inflammation of the body induced by a high-fat diet.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG1 is the results of the detection of serum 2-AAA levels in mice in each treatment group;

FIG2 is the results of the ALT level test in the serum of mice in each treatment group;

FIG3 is the test results of serum AST levels in mice in each treatment group;

FIG4 is the results of the detection of serum CRE levels in mice in each treatment group;

FIG5 is a representative picture of HE staining of liver and kidney of mice in each treatment group;

Figure 6 shows the positive expression of TNF-α and TGF-β in AS plaques of mice in each treatment group;

FIG7 is the test results of serum TNF-α levels in mice in each treatment group;

FIG8 is the test results of serum TGF-β levels in mice of each treatment group;

FIG9 is the detection result of TNF-α secretion by macrophages after 2-AAA treatment;

FIG10 is the detection result of TGF-β secretion by macrophages after 2-AAA treatment;

FIG11 is the mRNA expression level of P65 after 2-AAA treatment;

FIG. 12 shows the expression of proteins related to the NF-κB signaling pathway after 2-AAA treatment.

In the attached drawings, the "Chow" group described in the animal experiment section represents the control group receiving a standard diet, in which the mice not treated with 2-AAA are the "Chow+-" group, and the mice treated with 2-AAA are the "Chow+2-AAA" group. The "Westerndiet" group represents the group of mice receiving a high-fat diet, in which the mice not treated with 2-AAA are the "Western diet+-" group, and the mice treated with 2-AAA are the "Western diet+2-AAA" group. The "Con/Control" group described in the cell experiment section represents the control group, the "2-AAA" group represents the group treated with 2-AAA, the "ox-LDL" group represents the group treated with ox-LDL, and the "Combined" group represents the group treated with both 2-AAA and ox-LDL.

The drawings are only used for illustrative purposes and should not be construed as limitations on this patent. In order to better illustrate this embodiment, some parts of the drawings may be omitted, enlarged or reduced, and do not represent the size of the actual product. For those skilled in the art, it is understandable that some well-known structures and their descriptions in the drawings may be omitted. The same or similar numbers correspond to the same or similar parts. The terms describing the positional relationship in the drawings are only used for illustrative purposes and should not be construed as limitations on this patent.

Detailed ways

The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments in the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention. In addition, the technical solutions between the various embodiments can be combined with each other, but they must be based on the ability of ordinary technicians in this field to implement them. When the combination of technical solutions is contradictory or cannot be implemented, it should be considered that such a combination of technical solutions does not exist and is not within the scope of protection required by the present invention.

As an intermediate product in the lysine metabolic pathway, 2-AAA is mainly produced in the mitochondria of liver cells. Lysine first undergoes a condensation reaction with α-ketoglutarate through l-lysine-ketoglutarate reductase to form saccharin, which is then reduced to 2-aminoadipic acid semialdehyde by saccharin dehydrogenase, and finally converted to 2-AAA by 2-aminoadipic acid semialdehyde dehydrogenase. In the degradation metabolic pathway of 2-AAA, 2-AAA first undergoes a transamination reaction with α-ketoglutarate to form 2-ketoadipic acid, which is then decarboxylated by dehydrogenase to form glutaryl-CoA, and finally metabolized to acetyl-CoA through a series of CoA esters.

The present invention aims to solve the problem of chronic inflammation in the body caused by changes in lifestyle and eating habits. We propose a new product that inhibits the body's inflammatory response by regulating the levels of TNF-α and TGF-β. This product can regulate the body's chronic inflammation caused by lipid metabolism disorders, ultimately improving the body's metabolism and maintaining overall health.

The "Chow" group described in the following examples represents a control group receiving a standard diet, in which mice not treated with 2-AAA are the "Chow+-" group, and mice treated with 2-AAA are the "Chow+2-AAA" group. The "Western diet" group represents a group of mice receiving a high-fat diet, in which mice not treated with 2-AAA are the "Western diet+-" group, and mice treated with 2-AAA are the "Western diet+2-AAA" group. The "Con/Control" group described in the cell experiment section represents a control group, a "2-AAA" group represents a group treated with 2-AAA, a "ox-LDL" group represents a group treated with ox-LDL, and a "Combined" group represents a group treated with both 2-AAA and ox-LDL.

Example 1: Construction of a high-fat diet-induced atherosclerosis mouse model.

ApoE ^-/- mice (40) aged 6-8 weeks were purchased and randomly divided into a Chow group and a Western diet group. The Chow group was given a standard diet for 12 weeks, and the Western diet group was given a high-fat diet for 12 weeks.

The two groups of mice were treated by adding 2-AAA to their drinking water.

In this embodiment, 2-AAA was added to drinking water at a concentration of 2.2 g/L, and each group of mice had free access to drinking water. The mice were killed after 12 weeks of feeding with different diets, and samples were collected. After collecting the samples, the levels of 2-AAA, alanine aminotransferase (ALT), aspartate aminotransferase (AST), and creatinine (CRE) in serum were detected, and pathological experiments of atherosclerotic plaques in the liver, kidney, and aortic root were performed, as well as serum TNF-α and TGF-β levels were detected, and the results are shown in Figures 1 to 8.

FIG1 is the results of the detection of serum 2-AAA levels in mice in each treatment group.

The results showed that the addition of 2-AAA to drinking water significantly increased the 2-AAA content in serum of mice receiving a standard diet or a high-fat diet, indicating the effective administration of 2-AAA and the successful establishment of the model (*P<0.05, **P<0.01 compared with the corresponding control group).

Figure 2 shows the results of serum ALT level detection in mice of each treatment group. The results show that the intake of 2-AAA did not cause changes in serum ALT levels.

Figure 3 shows the results of serum AST level testing in mice of each treatment group. The results show that 2-AAA intake did not cause changes in serum AST levels.

Figure 4 shows the results of serum CRE level detection in mice of each treatment group. The results show that the intake of 2-AAA did not cause changes in serum CRE levels.

Figure 5 is a representative picture of HE staining of the liver and kidney of mice in each treatment group. By observing the pathological characteristics of the liver and kidney of mice in each treatment group, it was found that there was no obvious cell degeneration and inflammatory response in the liver and kidney tissues after taking 2-AAA.

In summary, combined with the results of Figures 2 to 5, it is preliminarily shown that the metabolism of 2-AAA in vivo has no obvious damaging effects on the liver and kidneys of mice, and exogenous 2-AAA treatment has been proven to be safe in in vivo experiments.

FIG6 shows the positive expression of TNF-α and TGF-β in AS plaques of mice in each treatment group.

The effect of 2-AAA on the expression levels of TNF-α and TGF-β in AS plaques of ApoE-/- mice was evaluated by immunohistochemistry. The experimental results showed that compared with the high-fat diet group, after consuming 2-AAA, the percentage of TNF-α positive area in AS plaques of ApoE-/- mice was significantly reduced, while the percentage of TGF-β positive area was significantly increased. This indicates that in the inflammatory environment of the body induced by a high-fat diet, 2-AAA has the effect of inhibiting the expression of TNF-α and promoting the expression of TGF-β, thereby inhibiting the inflammatory response (**P<0.01, ***P<0.001 compared with the corresponding control group).

Figure 7 is the test results of serum TNF-α levels in mice of each treatment group. The results showed that compared with the high-fat diet group, the level of inflammatory factor TNF-α in serum was significantly decreased after consuming 2-AAA (***P<0.001 compared with the corresponding control group).

Figure 8 is the test results of serum TGF-β levels in mice of each treatment group. The results showed that compared with the high-fat diet group, the level of anti-inflammatory factor TGF-β in serum was significantly increased after consuming 2-AAA (*P<0.05 compared with the corresponding control group).

The combined results of Figures 7-8 indicate that 2-AAA can affect the inflammatory level of ApoE-/- mice fed a high-fat diet.

Embodiment 2:

RAW264.7 macrophages were cultured and divided into Con/Control group, 2-AAA group, ox-LDL group and Combined group. The Con/Control group was used as the control, the 2-AAA group was treated with 2-AAA, the ox-LDL group was treated with ox-LDL, and the Combined group was treated with both 2-AAA and ox-LDL. The concentration of ox-LDL was 100 μg/mL; the concentration of 2-AAA was 200 mM. After treatment, the levels of TNF-α and TGF-β in the cell supernatant of each group were detected; the mRNA expression level of the P65 gene in each group of cells was detected, and the expression level of proteins related to the NF-κB signaling pathway in the cells was detected. The results are shown in Figures 9 to 12.

FIG. 9 is the detection result of TNF-α secretion by macrophages after 2-AAA treatment.

The results showed that ox-LDL could stimulate macrophages to secrete more TNF-α. Furthermore, compared with the ox-LDL group, in the Combined group, 2-AAA could inhibit the secretion of TNF-α induced by ox-LDL (##P＜0.01, ###P<0.001 compared with the ox-LDL group).

FIG. 10 is the detection result of TGF-β secretion by macrophages after 2-AAA treatment.

The results showed that ox-LDL treatment significantly reduced the secretion of TGF-β by macrophages. Furthermore, compared with the ox-LDL group, in the Combined group, 2-AAA could enhance the secretion of TGF-β by macrophages (###P<0.001 compared with the ox-LDL group).

FIG. 11 shows the mRNA expression level of P65 after 2-AAA treatment.

The results showed that compared with the control group, the mRNA expression level of the inflammation-related gene P65 was significantly reduced after treatment with 200mM 2-AAA. This indicates that 2-AAA can act on P65, a key transcription factor in the NF-κB signaling pathway (***P<0.001 compared with the control group).

FIG. 12 shows the expression of proteins related to the NF-κB signaling pathway after 2-AAA treatment.

The results showed that compared with the ox-LDL group, after adding 2-AAA, the expression level of P-IKB increased and the expression of P65 decreased significantly (##P＜0.01, ###P<0.001 compared with the ox-LDL group). Combined with the results of Figures 11 and 12, it is shown that 2-AAA can act on the NF-κB signaling pathway, a key signaling pathway for inflammation.

In summary, Figures 1 to 12 confirm that 2-AAA can reduce the secretion of TNF-α, promote the secretion of TGF-β, and inhibit the activation of the NF-κB signaling pathway, thereby inhibiting the high-fat-induced inflammatory response.

Finally, it should be emphasized that although we have described the technical solution of the present invention in detail in the preferred arrangement scheme, this is only used to illustrate the technical solution, not to limit it. Professionals in the technical field should understand that the technical solution of the present invention can be modified or replaced by equivalents. As long as it does not deviate from the core spirit and scope of the technical solution of the present invention, any improvement of the present invention, replacement of raw materials, addition of auxiliary components or selection of specific methods should be regarded as the content within the protection scope and disclosure scope of the present invention.

Claims (6)

1. An application of 2-aminoadipic acid in regulating inflammatory response induced by a high-fat diet, characterized in that the 2-aminoadipic acid is used to inhibit the production of tumor necrosis factor-α and promote transforming growth factor-β in serum. 2. The use of 2-aminoadipic acid in regulating the inflammatory response induced by a high-fat diet according to claim 1, characterized in that the 2-aminoadipic acid is used to inhibit the activation of the nuclear factor κB signaling pathway caused by oxidized low-density lipoprotein. 3. The use of 2-aminoadipic acid in regulating the inflammatory response induced by a high-fat diet according to claim 2, characterized in that the 2-aminoadipic acid is used to improve chronic inflammation caused by lipid metabolism disorders. 4. The use of 2-aminoadipic acid in regulating the inflammatory response induced by a high-fat diet according to claim 2, characterized in that the 2-aminoadipic acid is used to improve the inflammatory response of macrophages caused by high fat. 5. A food, medicine or health product for regulating chronic inflammation caused by a high-fat diet, characterized in that it contains 2-aminoadipic acid and/or its derivatives according to any one of claims 1 to 4. 6. The food, medicine or health product for regulating chronic inflammation caused by a high-fat diet according to claim 5, characterized in that the food, medicine or health product comprises an effective amount of 2-aminoadipic acid and/or its derivatives and auxiliary materials.