CN116617201A - Application of eicosatrienoic acid in regulating body inflammation - Google Patents

Abstract

The invention provides an application of eicosatrienoic acid in regulating body inflammation, and relates to the technical field of biomedicine. Eicosatrienoic acid (cis-11,14,17) (cis-11,14,17-Eicosatrienoic Acid, EA) of the present invention increases intestinal tight junction by regulating the gene expression of intestinal tight junction protein ZO-1 , inhibit white adipose tissue inflammation, and improve hepatic steatosis, and finally achieve the technical effect of improving the chronic inflammation of the body induced by high-fat diet.

Description

Application of eicosatrienoic acid in regulating body inflammation

technical field

The invention relates to the technical field of biomedicine, in particular to the application of eicosatrienoic acid in the treatment and or prevention of chronic inflammation in the body.

Background technique

Inflammation is a pathological condition caused by the body being stimulated by internal and external inflammatory factors and local damage. Inflammation is the basis of many chronic diseases, such as infectious diseases. If the cause is not removed, long-term chronic inflammation plays a role in promoting the induction, promotion, malignant transformation, invasion and metastasis of cancer. In addition, the mortality rate of related diseases caused by inflammation remains high globally, such as autoimmune diseases caused by chronic inflammation, tumors, aging, neurodegenerative or disordered diseases, and pathogens or microbial infections such as bacteria, fungi, parasites, and viruses wait. It can be said that all diseases are related to inflammation and are caused by inflammatory response.

Intestinal tissue is the largest mucosal immune site in the body. The intestinal mucosal epithelial barrier is the first line of defense against the invasion of pathogenic microorganisms and toxic substances in the intestinal tract. It refers to the sum of the structures and functions of the intestinal tract that can prevent certain harmful substances from entering other tissues, organs and blood circulation in the body through the intestinal mucosa. A normal and healthy intestinal mucosal barrier consists of three layers (mucus layer, epithelial glycoglobulin, and surface epithelial cells), and has the ability to block the invasion of pathogenic antigens, separate the intestinal lumen from the outside world, and maintain the ability to absorb nutrients. function. As an important part of the intestinal mucosal barrier, the intercellular junction complex is mainly composed of tight junctions (TJ), gap junctions (GJ), adherence junctions (AJ) and desmosomes. composition. As the most important connection between cells, tight junctions only allow small molecule soluble substances to pass through, and block the passage of toxic macromolecules and microorganisms. This special physiological function enables the normal physiological function of the intestinal barrier to be maintained. Tight junctions were first discovered because they can control permeability through the paracellular pathway. It is mainly composed of four transmembrane proteins (Occludin, Claudins, tricellulin, and JAM) and cytoplasmic proteins such as ZOs, cingulin, AF6, 7H6, rab3B, and symplekin. Molecules and cytoskeleton structures together constitute the tight junction protein complex, and its "zipper-like" anastomotic structure can effectively seal the top of the intercellular space, thereby preventing the invasion of toxic macromolecules. Once the integrity of the intestinal mucosal epithelial barrier is damaged, it will lead to a variety of intestinal diseases such as ulcerative colitis, Crohn's disease, chronic diarrhea and other inflammatory diseases of the body. As an important part of the mechanical barrier in the intestinal mucosal barrier, tight junction proteins have multiple functions. It can not only maintain the epithelial barrier function, block the invasion of toxic macromolecules and microorganisms, but also selectively regulate the entry of small molecules and ions into the body. In addition, tight junction proteins are also involved in the regulation of gene transcription, cell proliferation and differentiation status. As the three most important tight junction proteins, ZO-1 (zonula occludens 1), Occludin and Claudins are important proteins that constitute the integrity of the intestinal mucosal barrier and determine intestinal permeability.

Therefore, there is an urgent need for a food, medicine or health product that regulates chronic inflammation in the body by regulating the expression of intestinal tight junction proteins.

Contents of the invention

In view of the above problems, the present invention provides an application of eicosatrienoic acid in regulating chronic inflammation in the body, by providing a new product that inhibits inflammation in the body by increasing the tightness of the intestinal tract, the product regulates inflammation in the body while , which in turn can improve glucose and lipid metabolism disorders, and finally achieve the purpose of improving the body's metabolism.

In order to achieve the above object, a kind of application of eicosatrienoic acid in regulating chronic inflammation of body, eicosatrienoic acid (cis-11, 14, 17) (cis-11, 14, 17-Eicosatrienoic Acid, EA ) induces and/or increases the gene expression of intestinal tight junction protein ZO-1. Among them, intestinal tight junction protein is an important protein molecule that constitutes the intestinal mucosal barrier and determines intestinal permeability. The decrease of its expression can be regarded as an important sign of impaired intestinal barrier function.

Further, preferably, eicosatrienoic acid improves fatty degeneration of liver tissue and reduces the level of transaminase in liver tissue.

Further, preferably, eicosatrienoic acid reduces the expression of inflammatory proteins and genes in white adipose tissue; and eicosatrienoic acid reduces the level of lipopolysaccharide in the systemic circulation, wherein the administration of eicosatrienoic acid The amount is 1-1000 mg/kg body weight/day.

The invention also protects a drug, food or health product for regulating body inflammation, which contains eicosatrienoic acid and/or its derivatives.

Further, preferably, the medicine, food or health product contains an effective amount of eicosatrienoic acid and/or its derivatives and acceptable excipients.

The beneficial effects of the present invention are as follows:

The present invention aims at the current situation of frequent chronic inflammation of the body due to lifestyle and eating habits, and provides a new product that inhibits body inflammation by increasing intestinal tightness, eicosatrienoic acid (cis-11, 14, 17 )(cis-11, 14, 17-Eicosatrienoic Acid, EA) can increase intestinal tight junction, inhibit white adipose tissue inflammation, and improve hepatic steatosis by regulating the gene expression of intestinal tight junction protein ZO-1, and finally achieve improvement Technical effects of high-fat diet-induced chronic inflammation in the body.

Description of drawings

Hereinafter, the embodiments of the present invention will be described in conjunction with the accompanying drawings, wherein the "HFD" group represents the high-fat-induced mouse group fed with an equal volume of bovine serum albumin (Bovine Serum Albumin, BSA) saline solution, "HFD+EA" group means the high-fat-induced mouse group fed with EA (dissolved in BSA physiological saline solution) at a dose of 10 mg/kg body weight/day.

Figure 1 shows the detection results of the relative expression level of tight junction protein ZO-1 gene mRNA in the colon tissues of mice in the HFD and HFD+EA groups.

Figure 2 shows the detection results of lipopolysaccharide levels in the serum of mice in the HFD and HFD+EA groups.

A and B in Fig. 3 show the expression levels of each protein in the white adipose tissue of mice in the HFD and HFD+E groups and their digital results, respectively.

Figure 4 shows the detection results of the relative expression levels of each gene mRNA in the white adipose tissue of mice in the HFD and HFD+EA groups.

Figure 5 is the hematoxylin and eosin staining results of the liver tissues of mice in the HFD and HFD+EA groups.

A, B, C and D in Fig. 6 are the detection results of triglyceride, total cholesterol, alanine aminotransferase and aspartate aminotransferase in the livers of mice in the HFD and HFD+EA groups, respectively.

Figure 7 shows the detection results of fasting blood glucose levels of mice in the HFD and HFD+EA groups.

A and B in Fig. 8 show the detection results of glucose tolerance and the area under the glucose tolerance curve of mice in the HFD and HFD+EA groups, respectively.

A and B in Fig. 9 show the detection results of insulin tolerance and the area under the insulin tolerance curve of mice in the HFD and HFD+EA groups, respectively.

Detailed ways

The present invention will be more specifically illustrated on the basis of the following examples. However, the scope of protection of the present invention is not limited to the following examples.

The purpose of the present invention is to provide a new product that can inhibit the inflammation of the body by increasing the tightness of the intestinal tract, aiming at the current situation of chronic inflammation in the body caused by changes in lifestyle and eating habits. While regulating the chronic inflammation of the body, the product can In turn, it can improve the disorder of glucose and lipid metabolism, and finally achieve the purpose of improving the body's metabolism and maintaining the health of the matrix.

Eicosatrienoic acid (cis-11, 14, 17) (cis-11, 14, 17-Eicosatrienoic Acid, EA), the molecular formula is C ₂₀ H ₃₄ O ₂ ; it is a long-chain fatty acid. Its derivatives include but are not limited to fatty acid methyl esters, fatty acid ethyl esters, glycerides, fatty alcohols, fatty alcohol acetates, cholesterol esters; alkanol/enol mesylate, fatty acid soaps, wax esters and the like.

The "HFD" group described in the following examples represents the high-fat-induced mouse group fed with an equal volume of bovine serum albumin (Bovine SerumAlbumin, BSA) physiological saline solution, and the "HFD+EA" group represents the mouse group induced by 10 mg/ The high-fat-induced mouse group was fed with EA (dissolved in BSA physiological saline solution) at a dose of kg body weight/day. In the present invention, the HFD group and the HFD+EA group are statistically analyzed, and P<0.05 means that the difference between the two groups is statistically significant. Unless otherwise specified, the C57BL/6 mice used in the following examples were purchased from the Victorian Lihua Experimental Animal Center. All the reagents used were of analytical grade and commercially available from formal channels. The eicosatrienoic acid (cis-11, 14, 17) (cis-11, 14, 17-Eicosatrienoic Acid, EA) used was purchased from Shanghai Anpu Experiment Technology Co., Ltd.

It should be noted that in the specific implementation process, the high-fat-induced hyperlipidemia induced by EA (dissolved in BSA physiological saline solution) was fed with a dose of 1 mg/kg body weight/day to HFD and HFD+EA mice respectively. Group of mice; And, the group of mice induced by the high fat of EA (dissolved in BSA physiological saline solution) at the dosage of 1000mg/kg body weight/day; The experimental conclusion obtained is identical with the obtained conclusion of embodiment 1 .

Example 1

A high-fat diet-induced obesity mouse model was constructed.

20 4-week-old C57BL/6 mice were purchased and randomly divided into HFD group (10 mice) and HFD+EA group (10 mice), high-fat diet-induced obesity model.

The above two groups of mice were treated by intragastric administration.

The mice in the HFD+EA group were fed with eicosatrienoic acid (cis-11, 14, 17) (cis-11, 14, 17-Eicosatrienoic Acid, EA) daily at a dose of 10 mg/kg body weight/day, At the same time, the mice in the HFD group were fed with an equal volume of BSA saline solution. Mice were sacrificed after 17 weeks and samples were collected. During this period, weight was weighed every week, glucose tolerance was detected at week 15, insulin tolerance was detected after a week of rest, and triglyceride, total cholesterol, alanine aminotransferase and aspartate aminotransferase contents in liver tissue were detected after collecting samples, and Liver tissue staining experiments, white adipose tissue Western blot experiments and PCR experiments were performed. The results are shown in Figure 1-9. Among them, the "HFD" group represents the high-fat diet-induced mouse group fed with an equal volume of bovine serum albumin (Bovine SerumAlbumin, BSA) saline solution, and the "HFD+EA" group represents a dose of 10 mg/kg body weight/day The high-fat-induced mouse group fed with EA (dissolved in BSA saline solution).

Figure 1 shows the detection results of the relative expression level of tight junction protein ZO-1 gene mRNA in the colon tissues of mice in the HFD and HFD+EA groups. The results showed that daily administration of EA to mice fed a high-fat diet could significantly increase the gene expression of intestinal tight junction protein ZO-1, which indicated that EA treatment could increase intestinal tight junction (see Figure 1, "*" means p<0.05 ).

Fig. 2 is the detection result of lipopolysaccharide content in serum of HFD and HFD+EA two groups of mice. The results showed that daily administration of EA to mice fed a high-fat diet could significantly reduce the level of lipopolysaccharide in the circulation (see Figure 2, "***" means p<0.001).

Figure 3 A and B respectively show the expression levels of each protein in the white adipose tissue of mice in the HFD and HFD+EA groups and their data results. Inflammatory proteins include, but are not limited to, the pro-inflammatory protein NF-κB protein and the anti-inflammatory protein IκB-α. The results in Figure 3 show that EA treatment can reduce the expression of P-JNK protein by inhibiting the expression of TLR4 protein in white adipose tissue, thereby inhibiting the expression of pro-inflammatory protein NF-κB protein and promoting the expression of anti-inflammatory protein IκB-α. These data indicate that EA treatment can improve White adipose tissue inflammation (see Figure 3, "*" indicates p<0.05).

Figure 4 shows the detection results of the relative expression levels of each gene mRNA in the white adipose tissue of mice in the HFD and HFD+EA groups. The results showed that EA treatment significantly reduced the mRNA expression of pro-inflammatory cytokines MCP-1 gene and TNF-α gene in white adipose tissue (see Figure 4, "*" means p<0.05, "**" means p<0.01).

Figure 5 is the results of hematoxylin and eosin staining of the liver tissue sections of mice in the HFD and HFD+EA groups. The results showed that there was obvious fatty degeneration in the liver tissue of the mice fed the high-fat diet, and daily gavage of EA could improve the fatty degeneration of the liver tissue of the mice fed the high-fat diet (see FIG. 5 ).

A, B, C and D in Fig. 6 are the detection results of triglyceride, total cholesterol, alanine aminotransferase and aspartate aminotransferase in the livers of mice in the HFD and HFD+EA groups, respectively. The results showed that mice fed a high-fat diet could significantly reduce triglycerides, middle cholesterol, alanine aminotransferase and aspartate aminotransferase levels in liver tissue by intragastric administration of EA every day, which indicated that EA treatment could significantly improve liver lipid metabolism (see Figure 6, "*" means p<0.05, "**" means p<0.01).

Figure 7 shows the detection results of fasting blood glucose levels of mice in the HFD and HFD+EA groups. The results showed that daily gavage of EA in mice fed high-fat diet could significantly reduce fasting blood glucose level (see Figure 7, "*" means p<0.05).

A and B in Fig. 8 show the detection results of glucose tolerance and the area under the glucose tolerance curve of mice in the HFD and HFD+EA groups, respectively. The results showed that daily administration of EA to mice fed a high-fat diet could significantly reduce blood glucose levels at 15 minutes, 90 minutes and 120 minutes after glucose injection, and significantly reduce the area under the curve of the glucose tolerance test. These results prove that EA treatment can significantly improve Glucose tolerance (see Figure 8, "*" indicates p<0.05).

A and B in Fig. 9 show the detection results of insulin tolerance and the area under the insulin tolerance curve of mice in the HFD and HFD+EA groups, respectively. The results showed that daily administration of EA to mice fed a high-fat diet could significantly improve insulin tolerance and significantly reduce the area under the insulin tolerance curve. These results proved that EA treatment could significantly improve insulin sensitivity (see Figure 9, "*" indicates p<0.05, "**" means p<0.01).

As shown in Figures 1 to 9, the eicosatrienoic acid of the present invention can increase the gene expression of the intestinal tight junction protein ZO-1 by regulating the expression of the intestinal tight junction protein ZO-1, and can improve glucose and lipid metabolism disorders while regulating chronic inflammation in the body. To achieve the purpose of improving the body's metabolism and maintaining the health of the matrix. Wherein, the glucose and lipid metabolism disorder includes abnormal glucose and lipid metabolism caused by abnormal glucose and lipid metabolism, overweight, obesity, hyperlipidemia, hypertriglyceridemia, diabetes, fatty liver, atherosclerosis, hyperuricemia, high Blood pressure, coronary heart disease, cardiovascular and cerebrovascular diseases, kidney disease, polycystic ovary syndrome, glucagonemia, necrotic migratory erythema, glucagon tumors and other metabolic diseases, hypertensive heart disease, valvular Heart disease, alcoholic cardiomyopathy, cardiovascular complications of diabetes, or nerve damage diseases such as Alzheimer's disease or Parkinson's disease, polycystic ovary syndrome, etc.

In summary, Figures 1-9 confirm that the long-chain fatty acid - eicosatrienoic acid (cis-11, 14, 17) (cis-11, 14, 17-Eicosatrienoic Acid, EA) can regulate intestinal The gene expression of tight junction protein ZO-1 increases the intestinal tight junction, reduces the level of lipopolysaccharide in the systemic circulation, thereby inhibiting white adipose tissue inflammation, and improving liver lipid metabolism, thereby reducing the body's fasting blood sugar, improving the body's glucose tolerance and increasing the body's insulin Sensitivity, ultimately improving high-fat diet-induced glucose and lipid metabolism disorders. .

Among the various experimental supplies involved in this article (including but not limited to: chemical reagents, biological products, cells, organisms, instruments, etc.), for those that are special or not easy to obtain, the manufacturer, References or detailed preparation methods; without special instructions, are all routine experimental supplies, which can be easily obtained by various means (such as purchase, self-preparation, etc.) before the date of application.

Although, the present invention has been described in detail with general description, specific implementation and test above, but on the basis of the present invention, some modifications or improvements can be made to it, which will be obvious to those skilled in the art . Therefore, the modifications and improvements made on the basis of not departing from the spirit of the present invention all belong to the protection scope of the present invention.

Claims (4)

1. Use of eicosatrienoic acid for modulating inflammation in an organism, wherein eicosatrienoic acid induces and/or increases gene expression of the intestinal claudin ZO-1.

2. The use of eicosatrienoic acid as claimed in claim 1 to regulate inflammation in the body, wherein eicosatrienoic acid improves liver tissue steatosis and reduces the level of transaminase in liver tissue.

3. The use of eicosatrienoic acid as claimed in claim 1 to regulate inflammation in the body, wherein eicosatrienoic acid reduces the expression of inflammatory proteins and genes in white adipose tissue.

4. A medicament, food or health product for regulating inflammation of the body, characterized in that it comprises eicosatrienoic acid and/or its derivatives as claimed in any one of claims 1 to 3.