3-indolebutyric acid as a ppar gamma agonist for regulating lipid metabolism in fish

Abstract

The application belongs to the technical field of fish lipid metabolism, and discloses application of 3-indolebutyric acid as a PPAR gamma agonist for regulating fish lipid metabolism. The application discloses that IBA can be used as a safe and effective peroxisome proliferator-activated receptor gamma (PPAR gamma) activator, and can be used for regulating fish peroxisome proliferator-activated receptor signal pathways, insulin signal pathways, glyceride metabolism signal pathways, fatty acid biosynthesis signal pathways, fatty acid elongation signal pathways and other metabolic pathways, promoting absorption and conversion of lipid substances such as triglycerides in fish, promoting production and accumulation of lipid substances such as free fatty acids and polyunsaturated fatty acids in fish, and having the effect of promoting fish growth, and has a wide application prospect.

Description

Technical Field

[0001] This invention belongs to the field of fish lipid metabolism technology, specifically involving the application of 3-indolebutyric acid as a PPARγ agonist to regulate lipid metabolism in fish. Background Technology

[0002] Peroxisome proliferator-activated receptor gamma (PPARγ) is a crucial ligand-dependent transcription factor that influences physiological processes such as lipid metabolism, insulin sensitivity regulation, and adipocyte differentiation. PPARγ activation affects cell differentiation and proliferation, can lower triglyceride levels, regulate high-density lipoprotein (HDL) and low-density lipoprotein (LDL) levels, and is also essential for maintaining glycemic homeostasis, regulating inflammatory responses, and bone formation.

[0003] These functions provide a solid scientific basis for the development of PPARγ activators and point to their potential applications in the treatment of metabolic and inflammatory diseases. Several PPARγ agonists have been discovered and approved for the treatment of metabolic diseases, particularly diabetes, obesity, and related lipid metabolism disorders. Thiazolidinediones (including pioglitazone and rosiglitazone), which are already in use, effectively improve insulin sensitivity and lower blood glucose, and are used clinically to treat type 2 diabetes. In addition, several drugs that can activate PPARγ are under development or in the experimental stage, such as azelastine and atorvastatin. Studies have shown that certain natural metabolites can affect the differentiation process of adipocytes, the synthesis and oxidation of fatty acids, and the regulation of energy balance. For example, ginsenosides can exert their pharmacological effects by regulating glucose and lipid metabolism, and can activate the PPARγ signaling pathway, thereby promoting fatty acid utilization. However, many problems have been found in the clinical use of these drugs. Improper use or overuse of these drugs may lead to side effects such as weight gain, edema and fat accumulation, and even adverse effects on the skeletal and cardiovascular systems. In particular, these side effects may negatively affect the treatment effect in obese patients.

[0004] Therefore, there is an urgent need for a safe and reliable PPARγ agonist to regulate lipid metabolism and growth processes, and to develop novel bioactive agents for obesity and related diseases. Summary of the Invention

[0005] The first aspect of the present invention is to provide the use of 3-indolebutyric acid or its derivatives in the preparation of products that activate PPARγ.

[0006] A second aspect of the present invention aims to provide the use of 3-indolebutyric acid or its derivatives in the preparation of products that promote the expression of the ins, lpl, cel.1, dgat2, lida, elovl6, fads6, gh1 and / or igf2 genes.

[0007] A third aspect of the present invention aims to provide the use of 3-indolebutyric acid or its derivatives in the preparation of products that inhibit the expression of the acaca, fans and / or mstnb genes.

[0008] The fourth aspect of this invention aims to provide the use of 3-indolebutyric acid or its derivatives in the preparation of products that promote fish growth.

[0009] The fifth aspect of this invention aims to provide the use of 3-indolebutyric acid or its derivatives in the preparation of products that promote lipid metabolism.

[0010] The sixth aspect of this invention aims to provide a product.

[0011] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0012] This invention explores the activation effect of IBA on PPARγ in fish, as well as the biological functions and molecular mechanisms of IBA in fish lipid metabolism and individual growth through PPARγ activation. Experimental results show that IBA can regulate the expression of downstream insulin genes, lipid metabolism and growth-related genes by activating PPARγ, thereby affecting the conversion and utilization of lipids, especially triglycerides and polyunsaturated fatty acids, in fish and affecting fish growth.

[0013] A first aspect of the invention provides the use of 3-indolebutyric acid or a derivative thereof in the preparation of products that activate PPARγ.

[0014] In some embodiments of the present invention, the 3-indolebutyric acid or its derivatives include 3-indolebutyric acid or a pharmaceutically acceptable salt thereof.

[0015] In some embodiments of the present invention, the pharmaceutically acceptable salt includes at least one of a metal salt, an ammonium salt, a salt formed with an inorganic acid, a salt formed with an organic base, a salt formed with an organic acid, a salt formed with a basic amino acid, and a salt formed with an acidic amino acid.

[0016] In some embodiments of the present invention, the metal salt includes alkali metal salts and alkaline earth metal salts.

[0017] In some embodiments of the present invention, the alkali metal salt includes at least one of sodium salt and potassium salt.

[0018] In some embodiments of the present invention, the alkaline earth metal salt includes at least one of calcium salt, magnesium salt, barium salt, and aluminum salt.

[0019] In some embodiments of the present invention, the salt formed with an organic base includes at least one of the following organic bases: trimethylamine, triethylamine, pyridine, methylpyridine, 2,6-dimethylpyridine, ethanolamine, diethanolamine, triethanolamine, cyclohexylamine, dicyclohexylamine, and N,N'-dibenzylethylenediamine.

[0020] In some embodiments of the present invention, the salt formed with the inorganic acid includes at least one of the following inorganic acids: hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, and phosphoric acid.

[0021] In some embodiments of the present invention, the salt formed with the organic acid includes at least one of the following organic acids: formic acid, acetic acid, trifluoroacetic acid, phthalic acid, fumaric acid, oxalic acid, tartaric acid, maleic acid, citric acid, succinic acid, malic acid, methanesulfonic acid, benzenesulfonic acid, and p-toluenesulfonic acid.

[0022] In some embodiments of the present invention, the salt formed with the basic amino acid includes at least one of the following basic amino acids: arginine, lysine, and ornithine.

[0023] In some embodiments of the present invention, the salt formed with the acidic amino acid includes a salt formed with at least one of the following acidic amino acids: aspartic acid and glutamic acid.

[0024] In some embodiments of the present invention, the product includes a reagent or a drug.

[0025] In some embodiments of the present invention, the product includes pharmaceutically acceptable excipients and / or any one or more other active ingredients.

[0026] In some embodiments of the present invention, the pharmaceutically acceptable excipients include at least one of the following: solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, binders, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, flow aids, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesion agents, integrators, penetration enhancers, pH adjusters, buffers, plasticizers, surfactants, foaming agents, defoamers, thickeners, encapsulating agents, humectants, absorbents, diluents, flocculants and anti-flocculators, filter aids, release inhibitors, and carriers.

[0027] In some embodiments of the present invention, for ease of administration, the active ingredient 3-indolebutyric acid or a pharmaceutically acceptable salt thereof may be processed with one or more pharmaceutically acceptable excipients into a specific dosage form. These excipients may be diluents (e.g., starch, pregelatinized starch, dextrin, sucrose, lactose, mannitol, and microcrystalline cellulose), absorbents (e.g., calcium sulfate, dicalcium phosphate, light magnesium oxide, and calcium carbonate), wetting agents (e.g., water and ethanol), binders (e.g., hydroxypropyl methylcellulose, povidone, starch paste, and syrup), disintegrants (e.g., dry starch, sodium hydroxymethyl starch, low-substituted hydroxypropyl cellulose, effervescent disintegrants, and crospovidone), and lubricants (magnesium stearate, talc, hydrogenated vegetable oil, polyethylene glycol, and micronized powders). The following are examples of agents: silica gel, colorants (such as titanium dioxide, sunset yellow, methylene blue, and pharmaceutical iron oxide), coating materials (such as acrylic resin, hydroxypropyl methylcellulose, and povidone), solvents (such as water for injection, ethanol, propylene glycol, and glycerin), acid-base adjusters (such as hydrochloric acid, lactic acid, sodium hydroxide, tartaric acid, and sodium tartrate), antioxidants (such as sodium sulfite, sodium metabisulfite, and sodium thiosulfate), antibacterial agents (such as phenol, benzyl alcohol, and thimerosal), and isotonic adjusters (such as sodium chloride and glucose).

[0028] In some embodiments of the present invention, the dosage form of the drug includes a gastrointestinal dosage form or a non-gastrointestinal dosage form.

[0029] In some embodiments of the present invention, the gastrointestinal dosage form includes at least one of powder, tablet, granule, capsule, sustained-release, solution, dry suspension, effervescent tablet, emulsion, suspension, syrup, drops, and chewable tablet.

[0030] In some embodiments of the present invention, the gastrointestinal dosage forms include, but are not limited to, enteric-coated tablets, coated tablets, film-coated tablets, sugar-coated tablets, dispersible tablets, sucking tablets, chewable tablets, effervescent tablets, scratch tablets, sustained-release and controlled-release dosage forms, sustained-release tablets, sustained-release coated tablets, controlled-release tablets, orally disintegrating tablets, lozenges, and oral patches.

[0031] In some embodiments of the present invention, the non-gastrointestinal drug delivery dosage form includes at least one of injection dosage form, respiratory dosage form, skin dosage form, mucosal dosage form, and cavity dosage form.

[0032] In some embodiments of the present invention, the injectable dosage forms include, but are not limited to, injection solutions, solutions for injection, injection solutions for intravenous infusion, suspensions for injection, sterile powders for injection, intravenous injections, water injections, emulsions for injection, powder injections, injections, sterile powder injections, lyophilized powder injections, etc.

[0033] In some embodiments of the present invention, the effective concentration of 3-indolebutyric acid or its derivatives in the product is 0.2 to 5 μmol / mL.

[0034] A second aspect of the invention provides the use of 3-indolebutyric acid or a derivative thereof in the preparation of products that promote the expression of the ins, lpl, cel.1, dgat2, lida, elovl6, fads6, gh1 and / or igf2 genes.

[0035] In some embodiments of the present invention, the 3-indolebutyric acid or its derivatives promote the expression of the ins, lpl, cel.1, dgat2, lida, elovl6, fads6, gh1 and / or igf2 genes in vitro.

[0036] In some embodiments of the present invention, the 3-indolebutyric acid or its derivatives include 3-indolebutyric acid or a pharmaceutically acceptable salt thereof.

[0037] In some embodiments of the present invention, the pharmaceutically acceptable salt includes at least one of a metal salt, an ammonium salt, a salt formed with an inorganic acid, a salt formed with an organic base, a salt formed with an organic acid, a salt formed with a basic amino acid, and a salt formed with an acidic amino acid.

[0038] A third aspect of the invention provides the use of 3-indolebutyric acid or a derivative thereof in the preparation of products that inhibit the expression of the acaca, fans and / or mstnb genes.

[0039] In some embodiments of the present invention, the 3-indolebutyric acid or its derivatives inhibit the expression of the acaca, fans and / or mstnb genes in vitro.

[0040] In some embodiments of the present invention, the 3-indolebutyric acid or its derivatives include 3-indolebutyric acid or a pharmaceutically acceptable salt thereof.

[0041] In some embodiments of the present invention, the pharmaceutically acceptable salt includes at least one of a metal salt, an ammonium salt, a salt formed with an inorganic acid, a salt formed with an organic base, a salt formed with an organic acid, a salt formed with a basic amino acid, and a salt formed with an acidic amino acid.

[0042] A fourth aspect of the invention provides the use of 3-indolebutyric acid or a derivative thereof in the preparation of products that promote fish growth.

[0043] In some embodiments of the present invention, the 3-indolebutyric acid or its derivatives include 3-indolebutyric acid or a pharmaceutically acceptable salt thereof.

[0044] In some embodiments of the present invention, the pharmaceutically acceptable salt includes at least one of a metal salt, an ammonium salt, a salt formed with an inorganic acid, a salt formed with an organic base, a salt formed with an organic acid, a salt formed with a basic amino acid, and a salt formed with an acidic amino acid.

[0045] In some embodiments of the present invention, the fish includes zebrafish.

[0046] In some embodiments of the present invention, the 3-indolebutyric acid or its derivatives promote fish growth by regulating the expression of growth-promoting genes (gh1 and igf2 genes).

[0047] A fifth aspect of the invention provides the use of 3-indolebutyric acid or a derivative thereof in the preparation of products that promote lipid metabolism.

[0048] In some embodiments of the present invention, the 3-indolebutyric acid or its derivatives include 3-indolebutyric acid or a pharmaceutically acceptable salt thereof.

[0049] In some embodiments of the present invention, the pharmaceutically acceptable salt includes at least one of a metal salt, an ammonium salt, a salt formed with an inorganic acid, a salt formed with an organic base, a salt formed with an organic acid, a salt formed with a basic amino acid, and a salt formed with an acidic amino acid.

[0050] In some embodiments of the present invention, the 3-indolebutyric acid or its derivatives promote lipid metabolism by activating PPARγ, which regulates the main regulators of the insulin signaling pathway and overall insulin sensitivity.

[0051] A sixth aspect of the invention provides a product comprising 3-indolebutyric acid or a derivative thereof, and a PPARγ agonist.

[0052] In some embodiments of the present invention, the PPARγ agonist includes at least one of rosiglitazone, pioglitazone, azelastine, atorvastatin, ginsenosides, benzoates, and phenylacetates.

[0053] In some embodiments of the present invention, the product includes a drug.

[0054] In some embodiments of the present invention, the product can be used to regulate lipid metabolism and fish growth.

[0055] The beneficial effects of this invention are:

[0056] This invention discloses for the first time that IBA can be used as a safe and effective peroxisome proliferator-activated receptor γ (PPARγ) activator. It can be used to regulate metabolic pathways such as the peroxisome proliferator-activated receptor signaling pathway, insulin signaling pathway, glycerol metabolism signaling pathway, fatty acid biosynthesis signaling pathway, and fatty acid elongation signaling pathway in fish. It promotes the absorption and conversion of lipids such as triglycerides in fish, and promotes the production and accumulation of lipids such as free fatty acids and polyunsaturated fatty acids in fish. It has the effect of promoting fish growth and has broad application prospects. Attached Figure Description

[0057] Figure 1 The image shows the predicted 3D structure of the PPARγ protein, where (a) is a schematic diagram of the 3D structure and (b) is the prediction alignment error.

[0058] Figure 2 The results show the molecular docking of PPARγ protein and IBA, where (a) is the molecular model of IBA and (b) is the docking of PPARγ protein and IBA at the docking site with the lowest molecular docking energy.

[0059] Figure 3 The results show the safety assessment of IBA. A represents the culture status of zebrafish in the control group; B represents the culture status of zebrafish in the 0.2 μmol / mL IBA addition group; C represents the culture status of zebrafish in the 0.4 μmol / mL IBA addition group; D represents the culture status of zebrafish in the 0.8 μmol / mL IBA addition group; E represents the culture status of zebrafish in the 1 μmol / mL IBA addition group; and F shows that the mortality rate of zebrafish in the IBA addition group and the control group was similar.

[0060] Figure 4 Oil Red O staining results for zebrafish in different groups are shown. A to E are the Oil Red O staining results for zebrafish in the NC group, Add IBA group, Add agonist group, Add antagonist group, and Add IBA and agonist group, respectively.

[0061] Figure 5 To quantitatively analyze the results of Oil Red O staining and PUFA detection in each group of zebrafish using ImageJ software, (a) represents the relative area of ​​Oil Red, (b) represents the average optical density, and (c) represents the relative PUFA content. The letters in the dendrogram in the figure indicate significance (i.e., the same letter indicates no significant difference, while different letters indicate a significant difference).

[0062] Figure 6The pparγ(A) and ins(B) genes in zebrafish from different treatment groups are quantified. The letters in the dendrogram in the figure indicate significance (i.e., the same letter indicates no significant difference, while different letters indicate a significant difference).

[0063] Figure 7 The results of gene quantification for lpl (A), cel.1 (B) and dgat2 (C) in zebrafish from different treatment groups are shown in the figure. The letters in the dendrogram indicate significance (i.e., the same letter indicates no significant difference, while different letters indicate a significant difference).

[0064] Figure 8 The quantitative results of the lidea (A), acaca (B), and fans (C) genes in zebrafish from different treatment groups are shown in the figure. The letters in the dendrogram indicate significance (i.e., the same letter indicates no significant difference, while different letters indicate a significant difference).

[0065] Figure 9 The results of gene quantification of elovl6(A) and fads6(B) in zebrafish from different treatment groups are shown in the figure. The letters in the tree diagram indicate significance (i.e., the same letter indicates no significant difference, and different letters indicate significant difference).

[0066] Figure 10 The results of gene quantification for mstnb(A), gh1(B) and igf2(C) in zebrafish from different treatment groups are shown in the figure. The letters in the dendrogram indicate significance (i.e., the same letter indicates no significant difference, while different letters indicate significant difference). Detailed Implementation

[0067] The present invention will be further described in detail below through specific embodiments.

[0068] It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention.

[0069] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer shall apply. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased commercially.

[0070] The features and performance of the present invention will be further described in detail below with reference to embodiments.

[0071] Example 1: Molecular docking between PPARγ and IBA

[0072] Previous experiments have shown that 3-indolebutyric acid (IBA, molecular formula C) 12 H 13 NO2 (CAS No. 133-32-4) exerts its function through peroxisome β-oxidation, and activation of peroxisome proliferator-activated receptors (PPARs) promotes fatty acid β-oxidation within the peroxisome. Peroxisome β-oxidation acts as an intracellular fatty acid sensor, regulating lipolysis. These mechanisms suggest a possible link between IBA and PPARγ, and their potential role in regulating lipid metabolism. This embodiment investigates the existence of a link between IBA and PPARγ through molecular docking.

[0073] 1. Prediction of PPARγ protein structure

[0074] The sequence of the zebrafish PPARγ gene (SEQ ID NO:1) was obtained from NCBI (https: / / www.ncbi.nlm.nih.gov / ), and the amino acid sequence of the zebrafish PPARγ protein (SEQ ID NO:2) was obtained from the UniProt database (https: / / www.uniprot.org / ). The structure of the PPARγ protein was predicted using AlphaFold 3 (https: / / alphafoldserver.com / ), and a pdb format file was obtained.

[0075] PPARγ gene sequence:

[0076]

[0077] The amino acid sequence of the PPARγ protein:

[0078] (SEQ ID NO:2).

[0079] 2. Molecular docking between PPARγ and IBA

[0080] IBA's .mol ​​format file was obtained from ChemSpider (https: / / www.chemspider.com / ). Molecular docking between PPARγ and IBA was performed using Autodock (https: / / autodock.scripps.edu / ), and the docking energy between the two molecules was obtained. Post-docking visualization and processing were then performed using Pymol (https: / / pymol.org / ).

[0081] The results are as follows Figures 1-2 As shown, Figure 1 The predicted 3D structure of the PPARγ protein. Figure 2 (a) shows the molecular model of IBA, and Table 1 lists the 10 docking sites with the lowest molecular docking energies. Figure 2Image (b) shows the docking of PPARγ protein and IBA at the docking site with the lowest molecular docking energy. It can be observed that there are both hydrophobic and hydrogen bond interactions between PPARγ protein and IBA. The lowest docking energy between them is -6.17 kcal / mol, indicating the existence of a possible binding site between PPARγ protein and IBA.

[0082] Example 2: Safety Testing of IBA

[0083] Fifty healthy, germ-free zebrafish, 6 days post-fertilization (6 dpf), were randomly divided into four groups: IBA supplementation groups (0.2 μmol / mL, 0.4 μmol / mL, 0.8 μmol / mL, and 1.0 μmol / mL) and a control group (NC), with 10 zebrafish in each group. The IBA supplementation groups received a certain amount of IBA to achieve a concentration of 0.2 μmol / mL in the water, while the NC group received the same volume of physiological saline. The zebrafish were cultured continuously at 28°C and observed for 10 days, and mortality was recorded.

[0084] The results are as follows Figure 3 As shown, Figure 3 Figures A through E represent the culture status of the fish. The mortality rate of zebrafish was similar in the 0.2 μmol / mL group and the control group, with no significant difference (P = 0.4806). Figure 3 The concentration of F indicates that IBA is relatively safe at this concentration.

[0085] Example 3: Activation effect of IBA on PPARγ

[0086] After boiling the eggs, the egg yolks were mixed with sterile water at a ratio of 1:10 (m / m) to obtain a sterile egg yolk solution. IBA, the PPARG agonist rosiglitazone, and the PPARG inhibitor GW9662, dissolved in dimethyl sulfoxide (DMSO), were mixed with the egg yolk solution and shaken to mix thoroughly. The concentration of each additive in the mixture was 20 μmol / mL. This mixture was used as the stock solution for each additive and added to the zebrafish aquaculture water in proportion. The final concentration of the corresponding additive in the zebrafish aquaculture water for each additive group was 2 μmol / mL.

[0087] One hundred and twenty healthy, germ-free zebrafish, six days post-fertilization (6 dpf), were randomly divided into six groups: control (NC), IBA-added group (Add IBA), PPARG agonist-added group (Add agonist), PPARG inhibitor GW9662-added group (Add antagonist), and IBA and GW9662 mixed-feeding group (Add IBA and agonist). Zebrafish were collected as needed five hours after feeding for Oil Red O staining, polyunsaturated fatty acid (PUFA) determination, and gene quantification. Primers used for gene quantification are shown in Table 1.

[0088] Table 1. Quantitative analysis of genes and their primers

[0089]

[0090] Oil Red O staining results of zebrafish from different groups are as follows: Figure 4 As shown. NC group ( Figure 4 (A), Add IBA group ( Figure 4 (B) and Add agonist ( Figure 4 The staining results of the gastrointestinal tract of zebrafish in group C were quite similar, significantly different from those in the Add antagonist group. Figure 4 (D) and Add IBA and agonist group ( Figure 4 The stained area and color of the gastrointestinal tract of zebrafish (E) were larger. Add antagonist group ( Figure 4 (D) The staining area of ​​the gastrointestinal tract of zebrafish was very small and the color was very light. However, the addition of IBA could effectively restore the staining of the gastrointestinal tract of zebrafish with added PPARG inhibitors. Figure 4 (E). The Oil Red O staining of each group of zebrafish was quantitatively analyzed using ImageJ software. The results are as follows: Figure 5 As shown in (a) and (b), there were no significant differences in Oil Red staining area and mean optical density between the Add IBA group and the Addagonist group, but both were larger than those in the NC group. Adding IBA significantly increased the Oil Red staining area and mean optical density of zebrafish treated with the inhibitor. These results indicate that adding IBA can promote the absorption of triglycerides in zebrafish.

[0091] Polyunsaturated fatty acids (PUFAs) in zebrafish from different treatment groups were detected using a fish polyunsaturated fatty acid ELISA kit (Shanghai Huabang Biotechnology). Results are as follows: Figure 5As shown in (c), the zebrafish in the agonist group had the highest PUFA content, followed by the zebrafish in the IBA-treated group. The zebrafish in the inhibitor group had the lowest PUFA content, and the addition of IBA could increase the PUFA content in the zebrafish fed the inhibitor to some extent. The PUFA content detection results indicate that the addition of IBA can promote the production of PUFA in zebrafish.

[0092] PPARγ plays a central regulatory role in lipid and glucose metabolism, and is a major regulator of insulin signaling pathways and overall insulin sensitivity. Insulin encoded by the ins gene plays a crucial role in promoting lipid uptake and lipogenesis. Gene quantification results showed that the effects of IBA supplementation on zebrafish lipid metabolism and growth genes were similar to those of PPARγ agonist supplementation. IBA promoted the expression of both pparγ and ins genes, consistent with the effects of treatment with the PPARγ agonist rosiglitazone. Treatment with PPARγ inhibitors reduced the expression of both pparγ and ins genes. The combined use of IBA and PPARγ inhibitors not only restored the inhibitory effect of PPARγ inhibitors on the pparγ gene but also, to some extent, promoted pparγ gene expression. Figure 6 These results demonstrate that IBA effectively activates PPARγ and regulates the insulin signaling pathway.

[0093] The lpl gene in the triglyceride metabolism pathway catalyzes the hydrolysis of triglycerides in chylomicrons and very low-density lipoproteins, producing fatty acids and diacylglycerols for tissue use. The cel.1 gene participates in the hydrolysis and absorption of dietary fats, cholesterol esters, and fat-soluble vitamin cholesterol, while the dgat2 gene catalyzes the reaction of acyl-CoA with diacylglycerols to produce triglycerides. Experimental results showed that IBA promoted the expression of lpl, cel.1, and dgat2 genes, consistent with the expression trend after treatment with the PPARγ agonist rosiglitazone. IBA had a stronger promoting effect on lpl expression than rosiglitazone. PPARγ inhibitor treatment reduced the expression of lpl, cel.1, and dgat2 genes. The combined use of IBA and PPARγ inhibitors could, to some extent, restore the inhibitory effect of PPARγ inhibitors on lpl, cel.1, and dgat2 genes; however, the dgat2 gene still showed a significantly upregulated expression trend after the combined use. Figure 7 These results indicate that IBA can promote the absorption and conversion of lipids in the intestine, which is consistent with the results of Oil Red staining.

[0094] The *lipea* gene expression product, as a key rate-limiting enzyme in lipolysis, primarily functions to catalyze the breakdown of triglycerides stored in adipose tissue, releasing free fatty acids, including long-chain fatty acids, saturated fatty acids, and unsaturated fatty acids. These free fatty acids are an important energy source for the body. The *acaca* and *fasn* genes (signaling pathways for fatty acid biosynthesis) play a role in the conversion of long-chain fatty acids from malonyl-CoA. Experimental results showed that IBA had activating and inhibitory effects on the expression of the *lipea*, *acaca*, and *fans* genes, respectively, consistent with the expression trends after treatment with the PPARγ agonist rosiglitazone. Furthermore, IBA had a stronger promoting effect on *lipea* gene expression than rosiglitazone. Treatment with PPARγ inhibitors upregulated the expression of the *acaca* and *fans* genes. The combined use of IBA and PPARγ inhibitors not only inhibited the effect of PPARγ inhibitors on the expression of the *acaca* and *fans* genes but also significantly downregulated their expression. Figure 8 These results indicate that the addition of IBA reduces the production of long-chain fatty acids from malonyl-CoA, but increases the breakdown and conversion of triglycerides in adipose tissue, releasing free fatty acids that can provide energy for the body.

[0095] The elovl6 gene (fatty acid elongation signaling pathway) uses malonyl-CoA as a substrate and participates in the elongation of saturated and monounsaturated fatty acids. The fads6 gene (unsaturated fatty acid biosynthesis signaling pathway) has a desaturation function, converting these fatty acids into polyunsaturated fatty acids. The mstnb gene (cytokine-cytokine receptor interaction signaling pathway) has a negative regulatory effect on growth, while gh1 (growth hormone synthesis, secretion, and action signaling pathway) and IGF2 (MAPK signaling pathway) are important growth-promoting genes. Experimental results showed that IBA promoted the expression of both elovl6 and fads6 genes, consistent with the expression trend after treatment with the PPARγ agonist rosiglitazone, and the promoting effect of IBA on elovl6 gene expression was significantly higher than that of rosiglitazone. The combined use of IBA and a PPARγ inhibitor significantly inhibited the effect of the PPARγ inhibitor on the expression of elovl6 and fads6 genes, restoring their expression to normal levels. Figure 9 These results indicate that the addition of IBA can promote the production of saturated and unsaturated fatty acids, and ultimately promote the formation of PUFAs, which is consistent with the results of PUFA content detection.

[0096] IBA and the PPARγ agonist rosiglitazone had similar effects on the expression of mstnb, gh1, and igf2 genes; both inhibited mstnb gene expression and promoted gh1 and igf2 gene expression. There was no significant difference in the inhibitory effect of IBA and rosiglitazone on the mstnb gene, but the promoting effect of IBA on gh1 and igf2 gene expression was significantly lower than that of rosiglitazone. The combined use of IBA and a PPARγ inhibitor significantly affected the effect of the PPARγ inhibitor on the expression of mstnb and gh1 genes. Figure 10 These results indicate that adding IBA has a certain impact on growth, specifically, it can promote growth.

[0097] In summary, IBA is a safe and effective peroxisome proliferator-activated receptor gamma activator that can regulate the insulin signaling pathway in zebrafish, promote the absorption and conversion of lipids such as triglycerides, and promote the production and accumulation of lipids such as free fatty acids and polyunsaturated fatty acids, thus promoting the growth of zebrafish.

[0098] The embodiments of the present invention have been described in detail above with reference to the accompanying drawings. However, the present invention is not limited to the above embodiments, and various changes can be made within the scope of knowledge possessed by those skilled in the art without departing from the spirit of the present invention. Furthermore, the embodiments of the present invention and the features thereof can be combined with each other unless otherwise specified.

Claims

1,3-Indolebutyric acid or a pharmaceutically acceptable salt thereof in the preparation of products that promote lipid metabolism in fish; The promotion of lipid metabolism in fish refers to promoting the absorption and conversion of lipids in fish, and promoting the production and accumulation of lipids.

2. The application according to claim 1, characterized in that, The pharmaceutically acceptable salts include at least one of the following: metal salts, ammonium salts, salts formed with inorganic acids, salts formed with organic bases, salts formed with organic acids, salts formed with basic amino acids, and salts formed with acidic amino acids.

3. The application according to claim 2, characterized in that, The product also includes pharmaceutically acceptable excipients and / or any one or more active ingredients that activate PPARγ.

Images