Application of 3-indolebutyric acid as PPAR gamma agonist to regulation and control of fish lipid metabolism

Abstract

The invention belongs to the technical field of fish lipid metabolism, discloses application of 3-indolebutyric acid as a PPAR (peroxisome proliferator activated receptor) gamma agonist to regulation and control of fish lipid metabolism, and discloses that IBA can be used as a safe and effective PPAR gamma activator for the first time. The compound can be used for adjusting metabolic pathways such as a fish peroxisome proliferator-activated receptor signal pathway, an insulin signal pathway, a glyceride metabolism signal pathway, a biosynthesis signal pathway of fatty acid and a fatty acid extension signal pathway, and promoting absorption and conversion of fish to lipid substances such as triglyceride. The feed additive can promote the generation and accumulation of fat substances such as free fatty acids and polyunsaturated fatty acids of fishes, has the effect of promoting the growth of fishes, and has a wide application prospect.

Description

Technical Field

[0001] The invention belongs to the technical field of fish lipid metabolism, and particularly relates to application of 3-indolebutyric acid as a PPARγ agonist to regulate fish lipid metabolism. Background Art

[0002] Peroxisome proliferator-activated receptor γ (PPARγ) is a very important ligand-dependent transcription factor that affects physiological processes such as lipid metabolism, insulin sensitivity regulation, and adipocyte differentiation. The activation of PPARγ affects cell differentiation and proliferation, can reduce triglycerides, and regulate the content of high-density lipoprotein and low-density lipoprotein. It is also crucial for maintaining blood sugar homeostasis, regulating inflammatory responses, and bone formation.

[0003] These functions provide a solid scientific basis for the development of PPARG activators and point out their potential applications in the treatment of metabolic and inflammatory diseases. A variety of PPARγ agonists have been discovered and approved for the treatment of metabolic diseases, especially for the treatment of diabetes, obesity and related fat metabolism disorders. Thiazolidinediones (including pioglitazone and rosiglitazone) that have been used can effectively improve insulin sensitivity and lower blood sugar, and have been used in the clinical treatment of type 2 diabetes. In addition, there are some drugs that can activate PPARγ that are under development or in the experimental stage, such as Azevi and atorvastatin. Studies have shown that certain natural metabolites can affect the differentiation process of adipocytes, the synthesis and oxidation of fatty acids, the differentiation of adipocytes, and the regulation of energy balance. For example, ginsenosides can exert their pharmacological effects by regulating sugar and lipid metabolism, and can activate the PPARγ signaling pathway, thereby promoting the utilization of fatty acids. However, many problems have been found in the clinical use of the above-mentioned drugs. Improper use or excessive use of these drugs may cause side effects such as weight gain, edema, and fat accumulation, and even adverse effects on the bones and cardiovascular system. Especially in obese patients, these side effects may have a negative impact on the treatment effect.

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

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

[0006] The 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 ins, lpl, cel.1, dgat2, lipea, elovl6, fads6, gh1 and / or igf2 genes.

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

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

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

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

[0011] In order to achieve the above object, the technical solution adopted by the present invention is:

[0012] The present invention explores the activation effect of IBA on fish PPARγ, as well as the biological function and molecular mechanism of IBA in fish lipid metabolism and individual growth by activating PPARγ. The 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 lipid substances, especially triglycerides and polyunsaturated fatty acids, by fish, and affecting the growth of fish.

[0013] The first aspect of the present invention provides the use of 3-indolebutyric acid or its derivatives in the preparation of products for activating PPARγ.

[0014] In some embodiments of the present invention, the 3-indolebutyric acid or its derivative includes 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 an alkali metal salt or an alkaline earth metal salt.

[0017] In some embodiments of the present invention, the alkali metal salt includes at least one of a sodium salt and a 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 the organic base includes a salt formed with the following organic bases: at least one of trimethylamine, triethylamine, pyridine, picoline, 2,6-lutidine, ethanolamine, diethanolamine, triethanolamine, cyclohexylamine, dicyclohexylamine, and N,N'-dibenzylethylenediamine.

[0020] In some embodiments of the present invention, the salt formed with an inorganic acid includes a salt formed with the following inorganic acids: at least one of 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 a salt formed with the following organic acids: at least one of 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 a basic amino acid includes a salt formed with the following basic amino acids: at least one of 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 the following acidic amino acids: at least one of aspartic acid and glutamic acid.

[0024] In some embodiments of the present invention, the product comprises 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 solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, adhesives, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesives, integrators, penetration enhancers, pH regulators, buffers, plasticizers, surfactants, foaming agents, defoamers, thickeners, inclusion agents, humectants, absorbents, diluents, flocculants and deflocculating agents, filter aids, release retardants, and carriers.

[0027] In some embodiments of the present invention, in order to facilitate medication, the active ingredient 3-indolebutyric acid or its pharmaceutically acceptable salt can be processed into a specific dosage form with any one or more pharmaceutically acceptable excipients. These excipients can be diluents (such as starch, pregelatinized starch, dextrin, sucrose, lactose, mannitol and microcrystalline cellulose, etc.), absorbents (such as calcium sulfate, calcium hydrogen phosphate, light magnesium oxide and calcium carbonate, etc.), wetting agents (such as water and ethanol, etc.), binders (such as hydroxypropyl methylcellulose, povidone, starch slurry and syrup, etc.), disintegrants (such as dry starch, sodium hydroxymethyl starch, low-substituted hydroxypropyl cellulose, effervescent disintegrants and cross-linked polyvinylpyrrolidone, etc.), lubricants (magnesium stearate, talc, hydrogenated vegetable oil, polyethylene glycol and micropowder silica gel, etc.), colorants (such as titanium dioxide, sunset yellow, methylene blue and medicinal iron oxide, etc.), coating materials (such as acrylic resin, hydroxypropyl methylcellulose and povidone, etc.), solvents (such as water for injection, ethanol, propylene glycol and glycerol, etc.), acid-base regulators (such as hydrochloric acid, lactic acid, sodium hydroxide, tartaric acid and sodium tartrate, etc.), antioxidants (such as sodium sulfite, sodium pyrosulfite and sodium thiosulfate, etc.), antibacterial agents (such as phenol, benzyl alcohol and thimerosal, etc.), and isotonic regulators (such as sodium chloride and glucose, etc.).

[0028] In some embodiments of the present invention, the dosage form of the drug includes a dosage form for enteral administration or a dosage form for parenteral administration.

[0029] In some embodiments of the present invention, the dosage form for administration via the gastrointestinal tract includes at least one of powders, tablets, granules, capsules, sustained-release agents, solutions, dry suspensions, effervescent tablets, emulsions, suspensions, syrups, drops, and chewable tablets.

[0030] In some embodiments of the present invention, the dosage forms for administration through the gastrointestinal tract 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, scratched tablets, sustained-release and controlled-release dosage forms such as sustained-release tablets, sustained-release coated tablets, controlled-release tablets, orally disintegrating tablets, lozenges, oral patches, etc.

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

[0032] In some embodiments of the present invention, the injectable dosage forms include but are not limited to injection solutions, injection solutions, intravenous injection solutions, injection suspensions, sterile powders for injection, intravenous injections, water injections, injection emulsions, powder injections, injections, sterile powder injections, freeze-dried 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 0.2-5 μmol / mL.

[0034] The second aspect of the present invention provides the use of 3-indolebutyric acid or its derivatives in the preparation of products that promote the expression of ins, lpl, cel.1, dgat2, lipea, 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 ins, lpl, cel.1, dgat2, lipea, elovl6, fads6, gh1 and / or igf2 genes in vitro.

[0036] In some embodiments of the present invention, the 3-indolebutyric acid or its derivative includes 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] The third aspect of the present invention provides the use of 3-indolebutyric acid or its derivatives in the preparation of products for inhibiting the expression of 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 acaca, fans and / or mstnb genes in vitro.

[0040] In some embodiments of the present invention, the 3-indolebutyric acid or its derivative includes 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] The fourth aspect of the present invention provides the use of 3-indolebutyric acid or its derivatives in the preparation of products for promoting fish growth.

[0043] In some embodiments of the present invention, the 3-indolebutyric acid or its derivative includes 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 comprises zebrafish.

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

[0047] The fifth aspect of the present invention provides the use of 3-indolebutyric acid or its derivatives in the preparation of products promoting lipid metabolism.

[0048] In some embodiments of the present invention, the 3-indolebutyric acid or its derivative includes 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 activate PPARγ to regulate the insulin signaling pathway and the main regulatory factor of overall insulin sensitivity, thereby achieving the purpose of promoting lipid metabolism.

[0051] The sixth aspect of the present 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, azetidine, atorvastatin, ginsenoside, benzoate and phenylacetate.

[0053] In some embodiments of the invention, the product comprises a medicament.

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

[0055] The beneficial effects of the present invention are:

[0056] The present invention discloses for the first time that IBA can be used as a safe and effective peroxisome proliferator-activated receptor γ (PPARγ) activator, can be used to regulate metabolic pathways such as the peroxisome proliferator-activated receptor signaling pathway, the insulin signaling pathway, the glyceride metabolism signaling pathway, the fatty acid biosynthesis signaling pathway, the fatty acid elongation signaling pathway, etc. in fish, promotes the absorption and conversion of lipid substances such as triglycerides by fish, promotes the production and accumulation of lipid substances such as free fatty acids and polyunsaturated fatty acids in fish, has the effect of promoting fish growth, and has broad application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

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

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

[0059] Figure 3 The safety test results of IBA, where A is the culture condition of zebrafish in the control group, B is the culture condition of zebrafish in the 0.2μmol / mL IBA-added group, C is the culture condition of zebrafish in the 0.4μmol / mL IBA-added group, D is the culture condition of zebrafish in the 0.8μmol / mL IBA-added group, E is the culture condition of zebrafish in the 1μmol / mL IBA-added group, and F is that the mortality of zebrafish in the IBA-added group and the control group is similar.

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

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

[0062] Figure 6The quantitative results of pparγ (A) and ins (B) genes in zebrafish under different treatment groups. The letters in the dendrograms in the figure indicate significance (i.e., the same letters indicate no significant difference, and different letters indicate significant difference).

[0063] Figure 7 The quantitative results of lpl (A), cel.1 (B) and dgat2 (C) genes in zebrafish of different treatment groups. The letters in the dendrogram in the figure indicate significance (i.e., the same letters indicate no significant difference, and different letters indicate significant difference).

[0064] Figure 8 The quantitative results of lipea (A), acaca (B) and fans (C) genes in zebrafish under different treatment groups. The letters in the dendrogram in the figure indicate significance (i.e., the same letters indicate no significant difference, and different letters indicate significant difference).

[0065] Fig. 9 The quantitative results of elovl6 (A) and fads6 (B) genes in zebrafish under different treatment groups. The letters in the dendrograms in the figure indicate significance (i.e., the same letters indicate no significant difference, and different letters indicate significant difference).

[0066] Fig.10 The quantitative results of mstnb (A), gh1 (B) and igf2 (C) genes in zebrafish under different treatment groups. The letters in the dendrograms in the figure indicate significance (i.e., the same letters indicate no significant difference, and different letters indicate significant difference). DETAILED DESCRIPTION

[0067] The present invention is further described in detail below through specific examples.

[0068] It should be understood that these embodiments are only used to illustrate the present invention and are not used to limit the scope of the present invention.

[0069] In order to make the purpose, technical scheme and advantages of the embodiments of the present invention clearer, the technical scheme in the embodiments of the present invention will be described clearly and completely below. If the specific conditions are not specified in the embodiments, they are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be purchased commercially.

[0070] The features and performance of the present invention are further described in detail below in conjunction with the embodiments.

[0071] Example 1 Molecular docking between PPARγ and IBA

[0072] Preliminary experiments have shown that 3-indolebutyric acid (Indolebutyric acid, IBA, molecular formula C 12 H 13 NO2, CAS number: 133-32-4) can play a role through the peroxisome β-oxidation process, and the activation of peroxisome proliferator-activated receptors (PPARs) can promote the β-oxidation of fatty acids in the peroxisome. Peroxisome β-oxidation is a sensor of fatty acids in cells and can regulate fat decomposition. These mechanisms indicate that there may be a certain connection between IBA and PPARγ and that they play a role in regulating lipid metabolism. This example investigates whether there is a connection between IBA and PPARγ through molecular docking.

[0073] 1. Prediction of PPARγ protein structure

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

[0075] PPARγ gene sequence:

[0076]

[0077] Amino acid sequence of PPARγ protein:

[0078] (SEQ ID NO:2).

[0079] 2. Molecular docking between PPARγ and IBA

[0080] The mol format file of IBA was obtained from chemspider (https: / / www.chemspider.com / ). The molecular docking between PPARγ and IBA was performed using the software Autodock (https: / / autodock.scripps.edu / ) to obtain the molecular docking energy between the two molecules, and the software pymol (https: / / pymol.org / ) was used for visualization analysis and processing after docking.

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

[0082] Example 2 Safety Testing of IBA

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

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

[0085] Example 3 Activation effect of IBA on PPARγ

[0086] After the eggs are cooked, the egg yolks are mixed with sterile water at a ratio of 1:10 (m / m) to obtain a sterile egg yolk solution. IBA, PPARG agonist rosiglitazone, and PPARG inhibitor GW9662 dissolved in dimethyl sulfoxide (DMSO) are mixed with the egg yolk solution respectively, and oscillated to mix well. The concentration of various additives in the mixed solution is adjusted to 20 μmol / mL, and the mother solution of various additives is used as the mother solution of various additives, which are added to the zebrafish breeding water in proportion. The final concentration of the corresponding additives in each additive group in the zebrafish breeding water is 2 μmol / mL.

[0087] 120 healthy germ-free zebrafish at 6 days post fertilization (6 dpf) were divided into 6 groups, namely control group (NC), IBA-added group (Add IBA), PPARG agonist rosiglitazone-added group (Add agonist), PPARG inhibitor GW9662 group (Add antagonist), and IBA and GW9662 mixed feeding group (Add IBAand agonist). Zebrafish were collected as needed at 5 hours after feeding for Oil Red O staining, polyunsaturated fatty acid (PUFA) determination, and gene quantitative analysis. The primers used for gene quantification are shown in Table 1.

[0088] Table 1 Quantitative analysis genes and their primers

[0089]

[0090] The results of Oil Red O staining of zebrafish in different groups are shown in Figure 2. Figure 4 NC group ( Figure 4 Middle A), Add IBA group ( Figure 4 B) and Add agonist( Figure 4 The staining results of the gastrointestinal tract of zebrafish in the middle (C) group were similar, and significantly higher than those in the Add antagonist group ( Figure 4 D) and Add IBA and agonist group ( Figure 4 Middle E) The stained area of ​​zebrafish gastrointestinal tract is larger and the color is darker. Add antagonist group ( Figure 4 Middle D) The staining area of ​​zebrafish gastrointestinal tract is very small and the color is very light, while the addition of IBA can effectively restore the staining of zebrafish gastrointestinal tract with the addition of PPARG inhibitor ( Figure 4 ImageJ software was used to quantitatively analyze the Oil Red O staining of zebrafish in each group. Figure 5 As shown in (a) and (b), there was no significant difference in the oil red staining area and average optical density between the Add IBA group and the Addagonist group, but both were larger than the NC group. Adding IBA can significantly increase the oil red staining area and average optical density of zebrafish treated with inhibitors. These results indicate that adding IBA can promote the absorption of triglycerides by zebrafish.

[0091] The polyunsaturated fatty acids (PUFA) in zebrafish of different treatment groups were detected using the fish polyunsaturated fatty acid ELISA detection kit (Shanghai Huabang Biotechnology). Figure 5As shown in (c), the zebrafish in the agonist group had the highest PUFA content, followed by the zebrafish in the IBA 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 with the inhibitor to a certain extent. The PUFA content test results show that the addition of IBA can promote the production of PUFA in zebrafish.

[0092] PPARγ plays a core regulatory role in lipid metabolism and glucose metabolism. It is the main regulatory factor controlling the insulin signaling pathway and overall insulin sensitivity. The insulin encoded by the ins gene plays an important role in promoting lipid uptake and lipogenesis. Gene quantification results showed that the effects of adding IBA on zebrafish lipid metabolism and growth genes were similar to those of adding PPARγ agonists. IBA can promote the expression of pparγ and ins genes, which is consistent with the effect after treatment with the PPARγ agonist rosiglitazone. PPARγ inhibitor treatment reduces the expression of pparγ and ins genes. The mixed use of IBA and PPARγ inhibitors can not only restore the inhibitory effect of PPARγ inhibitors on the pparγ gene, but also promote the expression of the pparγ gene to a certain extent ( Figure 6 ). These results indicate that IBA can effectively activate PPARγ and regulate the insulin signaling pathway.

[0093] The lpl gene in the glyceride metabolism pathway signal catalyzes the hydrolysis of triglycerides in chylomicrons and very low-density lipoproteins to produce fatty acids and diacylglycerols for tissue utilization. The cel.1 gene is involved in the hydrolysis and absorption of fat, cholesterol esters and fat-soluble vitamin cholesterol in food, while the dgat2 gene can catalyze acyl-CoA and diacylglycerol to produce triglycerides. The experimental results showed that IBA can promote the expression of lpl, cel.1, and dgat2 genes, which is consistent with the expression trend after treatment with the PPARγ agonist rosiglitazone. IBA has a stronger promoting effect on lpl expression than rosiglitazone. PPARγ inhibitor treatment reduces the expression of lpl, cel.1, and dgat2 genes. The mixed use of IBA and PPARγ inhibitors can restore the inhibitory effect of PPARγ inhibitors on lpl, cel.1, and dgat2 genes to a certain extent. After mixed use, the dgat2 gene still shows a significant upregulated expression trend ( Figure 7 ). These results indicate that IBA can promote the absorption and transformation of lipid substances in the intestine, which is consistent with the results of oil red staining.

[0094] The expression product of the lipea gene is a key rate-limiting enzyme in the process of lipolysis. Its main function is to catalyze the decomposition of triglycerides stored in adipose tissue and release free fatty acids including long-chain fatty acids, saturated fatty acids, unsaturated fatty acids and other types of fatty acids. These free fatty acids are important energy sources for the body. The acaca gene and the fasn gene (fatty acid biosynthesis signaling pathway) play a role in the process of generating long-chain fatty acids from malonyl-CoA. The experimental results showed that IBA activated and inhibited the expression of lipea, acaca and fans genes, respectively, which was consistent with the expression trend after treatment with the PPARγ agonist rosiglitazone, and IBA's promoting effect on lipea gene expression was stronger than rosiglitazone. Treatment with PPARγ inhibitors increased the expression of acaca and fans genes. The mixed use of IBA and PPARγ inhibitors not only inhibited the effect of PPARγ inhibitors on the expression of acaca and fans genes, but also significantly downregulated the expression of acaca and fans genes ( Figure 8 ). These results indicate that the addition of IBA reduces the long-chain fatty acids generated from malonyl-CoA, but increases the decomposition 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 effect and can convert these fatty acids into polyunsaturated fatty acids. The mstnb gene (cytokine-cytokine receptor interaction signaling pathway) has a negative growth-regulating effect, while the gh1 (growth hormone synthesis, secretion and action signaling pathway) and igf2 gene (MAPK signaling pathway) are important growth-promoting genes. The experimental results showed that IBA promoted the expression of both elovl6 and fads6 genes, which was consistent with the expression trend after treatment with the PPARγ agonist rosiglitazone, and the promoting effect of IBA on the expression of the elovl6 gene was significantly higher than that of rosiglitazone. The combined use of IBA and PPARγ inhibitors significantly inhibited the effects of PPARγ inhibitors on the expression of elovl6 and fads6 genes, and restored the expression of elovl6 and fads6 genes to normal levels ( Fig. 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 PUFA, which is consistent with the results of PUFA content detection.

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

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

[0098] The embodiments of the present invention are described in detail above in conjunction with the accompanying drawings, but the present invention is not limited to the above embodiments. Various changes can be made within the knowledge of ordinary technicians in the relevant technical field without departing from the purpose of the present invention. In addition, the embodiments of the present invention and the features in the embodiments can be combined with each other without conflict.

Claims

1. 3-Application of indolebutyric acid or its derivatives in the preparation of products for activating PPARγ. 2.3-Use of indolebutyric acid or its derivatives in the preparation of products for promoting the expression of ins, lpl, cel.1, dgat2, lipea, elovl6, fads6, gh1 and / or igf2 genes.

3. Use of 3-indolebutyric acid or its derivatives in the preparation of products for inhibiting the expression of acaca, fans and / or mstnb genes. 4.3-Use of indolebutyric acid or its derivatives in the preparation of products for promoting fish growth. 5.3-Application of indolebutyric acid or its derivatives in the preparation of products promoting lipid metabolism.

6. The use according to any one of claims 1 to 5, characterized in that: The 3-indolebutyric acid or its derivatives include 3-indolebutyric acid or a pharmaceutically acceptable salt thereof.

7. The use according to claim 6, characterized in that: 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.

8. The use according to claim 7, characterized in that: The product also includes pharmaceutically acceptable excipients, and / or any one or more active ingredients capable of activating PPARγ.

9. The use according to claim 8, characterized in that: The pharmaceutically acceptable excipients include at least one of solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, adhesives, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesive agents, integrities, penetration enhancers, pH regulators, buffers, plasticizers, surfactants, foaming agents, defoamers, thickeners, inclusion agents, humectants, absorbents, diluents, flocculants and deflocculating agents, filter aids, release retardants, and carriers.

10. A product comprising 3-indolebutyric acid or a derivative thereof, and a PPARγ agonist; Preferably, the PPARγ agonist includes at least one of rosiglitazone, pioglitazone, ezetimibe, atorvastatin, ginsenoside, benzoate and phenylacetate.

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