Method for increasing content of melatonin in pig blood
By applying tryptophan or its salt to pigs, the problem of increasing the melatonin content in pigs is solved, and the level of melatonin is stable, and the pig's reproductive ability and pregnancy maintenance ability are enhanced.
Patent Information
- Application Number
- CN202411805315.3
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-09
- Publication Date
- 2025-03-07
AI Technical Summary
There has not been a simple and effective method to increase the melatonin content in pig blood, limiting its application in animal husbandry production.
The melatonin content in the blood of pigs is increased by administering tryptophan or a pharmaceutically acceptable salt thereof to the pig, using tryptophan as a substrate for melatonin synthesis. The application methods include oral, nasal, inhalation, etc., and the dosage and time of administration can be adjusted according to specific needs.
By administering tryptophan or its salt, the melatonin content in the pig's blood can be significantly improved, and by adjusting the amount and time of administration, the melatonin level can be stably improved, enhancing the pig's reproductive ability and pregnancy maintenance ability.
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Figure HDA0005180294110000012
Abstract
Description
Technical Field
[0001] The invention belongs to the field of animal husbandry and veterinary medicine, and in particular relates to a method for increasing the melatonin content in pig blood. Background Art
[0002] Melatonin (MT) is an indoleamine hormone mainly synthesized and secreted by the pineal gland. It has multiple biological functions, including regulating circadian rhythm, anti-oxidation, anti-inflammation and regulating the body's immunity. At present, a large number of literatures have shown that melatonin can improve the reproductive capacity of pigs by regulating multiple stages of pig reproductive activities. Under heat stress conditions, melatonin can alleviate the thermal damage of pig testicular supporting cells by promoting the expression of multiple antioxidant genes and regulating glucose metabolism. In vitro experiments have found that the addition of appropriate concentrations of melatonin can increase the level of progesterone synthesis in pig luteal cells, indicating that melatonin plays an important role in maintaining pig pregnancy. In addition, supplementing appropriate doses of melatonin in the late stage of pig pregnancy can effectively promote the expression of multiple antioxidant genes in placental tissue and increase the birth weight and weaning weight of piglets. At present, although a large amount of experimental data has shown that melatonin plays an important role in pig reproductive activities, melatonin has not yet passed veterinary drug approval, which limits its application in animal husbandry production.
[0003] Therefore, in view of the problems existing in the prior art, it is necessary to find a simple and effective method to increase the level of melatonin in pigs, which is crucial to the development of the pig industry.
[0004] Tryptophan, also known as β-indolylalanine, is one of the body's essential amino acids and a substrate for melatonin synthesis. It is reported that tryptophan can synthesize melatonin through hydroxylation, decarboxylation, transacetylation and transmethylation. In addition, tryptophan is a permitted feed additive. However, there are no reports on whether tryptophan supplementation can successfully increase melatonin levels in pigs. Summary of the invention
[0005] The first aspect of the present invention aims to provide a method for increasing the melatonin content in pig blood.
[0006] The second aspect of the present invention aims to provide the use of tryptophan or a pharmaceutically acceptable salt thereof.
[0007] In order to achieve the above object, the technical solution adopted by the present invention is:
[0008] In a first aspect, the present invention provides a method for increasing the melatonin content in pig blood, comprising administering tryptophan or a salt thereof to the pig.
[0009] In some embodiments, the administration of tryptophan or its salt comprises oral, nasal, inhalation, parenteral, intravenous, intraperitoneal, subcutaneous, intramuscular, intradermal, topical or rectal administration; further oral.
[0010] In some embodiments, the administration amount of tryptophan or its salt is 0.025-0.3 g / (Kg pig.day); further 0.04-0.26 g / (Kg pig.day); further 0.05-0.25 g / (Kg pig.day).
[0011] In some embodiments, the tryptophan or a salt thereof is administered 1-3 times / day; further once / day.
[0012] In some embodiments, the tryptophan or a salt thereof is administered for 1-6 days; further for 2-4 days.
[0013] In some embodiments, the administration amount of tryptophan or its salt is 0.24-0.26 g / (Kg pig.day) (preferably 0.25-0.26 g / (Kg pig.day)), and the administration days of tryptophan or its salt are 1-3 days (preferably 1-2 days); or
[0014] The administration amount of the tryptophan or its salt is 0.14-0.16 g / (Kg pig.day) (preferably 0.15-0.16 g / (Kg pig.day)), and the administration days of the tryptophan or its salt are 1-3 days (preferably 1-2 days); or
[0015] The administration amount of tryptophan or its salt is 0.04-0.06 g / (Kg pig.day) (preferably 0.05-0.06 g / (Kg pig.day)), and the administration days of tryptophan or its salt are 1-5 days (preferably 3-5 days; more preferably 3-4 days).
[0016] In some embodiments, the tryptophan or its salt is administered every morning (preferably between 7:00 and 8:00 a.m. every morning).
[0017] In some embodiments, the tryptophan or a salt thereof is added to feed for administration to pigs.
[0018] In some embodiments, the pig is a female pig.
[0019] In some embodiments, increasing the melatonin content in pig blood is increasing the melatonin content in pig peripheral blood.
[0020] In some embodiments, the increasing the melatonin content in pig blood is to stably increase the melatonin content in pig peripheral blood.
[0021] In some embodiments, the method does not involve diagnosis, and / or therapy.
[0022] In some embodiments, the salt comprises at least one of a metal salt, an ammonium salt, a salt formed with an organic base, a salt formed with an inorganic acid, a salt formed with an organic acid, a salt formed with a basic amino acid, and a salt formed with an acidic amino acid.
[0023] In some embodiments, the metal salt comprises at least one of an alkali metal salt (eg, sodium salt, potassium salt, etc.), an alkaline earth metal salt (eg, calcium salt, magnesium salt, barium salt, etc.), and an aluminum salt.
[0024] In some embodiments, the salt formed with an organic base comprises a salt formed with one or more of the following organic bases: trimethylamine, triethylamine, pyridine, picoline, 2,6-lutidine, ethanolamine, diethanolamine, triethanolamine, cyclohexylamine, dicyclohexylamine, and N,N'-dibenzylethylenediamine.
[0025] In some embodiments, the salt formed with an inorganic acid comprises a salt formed with one or more of the following inorganic acids: hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid.
[0026] In some embodiments, the salt formed with an organic acid comprises a salt formed with one or more 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.
[0027] In some embodiments, the salt formed with a basic amino acid comprises a salt formed with one or more of the following basic amino acids: arginine, lysine, ornithine.
[0028] In some embodiments, the salt formed with the acidic amino acid comprises a salt formed with one or more of the following acidic amino acids: aspartic acid, glutamic acid.
[0029] The second aspect of the present invention provides the use of tryptophan or its salt in a1) or a2):
[0030] a1) Increase the melatonin content in pig blood;
[0031] a2) preparing a product for increasing the melatonin content in pig blood.
[0032] In some embodiments, the product further comprises: instructions, wherein the instructions record: the method for increasing the melatonin content in pig blood according to the first aspect of the present invention.
[0033] In some embodiments, the product further comprises: other active ingredients that increase the melatonin content in pig blood.
[0034] In some embodiments, the product is a feed additive or a feed.
[0035] In some embodiments, the use described in a1) does not involve diagnosis and / or treatment.
[0036] In some embodiments, the use described in a1) is: administering tryptophan or a salt thereof to pigs.
[0037] In some embodiments, the administration of tryptophan or its salt comprises oral, nasal, inhalation, parenteral, intravenous, intraperitoneal, subcutaneous, intramuscular, intradermal, topical or rectal administration; further oral.
[0038] In some embodiments, the administration amount of tryptophan or its salt is 0.025-0.3 g / (Kg pig.day); further 0.04-0.26 g / (Kg pig.day); further 0.05-0.25 g / (Kg pig.day).
[0039] In some embodiments, the tryptophan or a salt thereof is administered 1-3 times / day; further once / day.
[0040] In some embodiments, the tryptophan or a salt thereof is administered for 1-6 days; further for 2-4 days.
[0041] In some embodiments, the administration amount of tryptophan or its salt is 0.24-0.26 g / (Kg pig.day) (preferably 0.25-0.26 g / (Kg pig.day)), and the administration days of tryptophan or its salt are 1-3 days (preferably 1-2 days); or
[0042] The administration amount of the tryptophan or its salt is 0.14-0.16 g / (Kg pig.day) (preferably 0.15-0.16 g / (Kg pig.day)), and the administration days of the tryptophan or its salt are 1-3 days (preferably 1-2 days); or
[0043] The administration amount of tryptophan or its salt is 0.04-0.06 g / (Kg pig.day) (preferably 0.05-0.06 g / (Kg pig.day)), and the administration days of tryptophan or its salt are 1-5 days (preferably 3-5 days; more preferably 3-4 days).
[0044] In some embodiments, the tryptophan or its salt is administered every morning (preferably between 7:00 and 8:00 a.m. every morning).
[0045] In some embodiments, the tryptophan or a salt thereof is added to feed for administration to pigs.
[0046] In some embodiments, the pig is a female pig.
[0047] In some embodiments, increasing the melatonin content in pig blood is increasing the melatonin content in pig peripheral blood.
[0048] In some embodiments, the increasing the melatonin content in pig blood is to stably increase the melatonin content in pig peripheral blood.
[0049] In some embodiments, the salt comprises at least one of a metal salt, an ammonium salt, a salt formed with an organic base, a salt formed with an inorganic acid, 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, the metal salt comprises at least one of an alkali metal salt (eg, sodium salt, potassium salt, etc.), an alkaline earth metal salt (eg, calcium salt, magnesium salt, barium salt, etc.), and an aluminum salt.
[0051] In some embodiments, the salt formed with an organic base comprises a salt formed with one or more of the following organic bases: trimethylamine, triethylamine, pyridine, picoline, 2,6-lutidine, ethanolamine, diethanolamine, triethanolamine, cyclohexylamine, dicyclohexylamine, and N,N'-dibenzylethylenediamine.
[0052] In some embodiments, the salt formed with an inorganic acid comprises a salt formed with one or more of the following inorganic acids: hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid.
[0053] In some embodiments, the salt formed with an organic acid comprises a salt formed with one or more 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.
[0054] In some embodiments, the salt formed with a basic amino acid comprises a salt formed with one or more of the following basic amino acids: arginine, lysine, ornithine.
[0055] In some embodiments, the salt formed with the acidic amino acid comprises a salt formed with one or more of the following acidic amino acids: aspartic acid, glutamic acid.
[0056] The beneficial effects of the present invention are:
[0057] The invention provides a method for increasing the melatonin content in pig blood. Tryptophan or its salt is administered to pigs. The melatonin content in the pig blood can be increased by administering tryptophan or its salt to pigs.
[0058] Furthermore, by limiting the daily administration amount and administration days of tryptophan or its salt, the melatonin content in pig blood can be stably increased.
[0059] Furthermore, by adding tryptophan or its salt into feed and administering it to pigs, the method of the present invention is simple and easy to operate, will not cause stress response in pigs, and has strong feasibility.
[0060] The method for increasing the melatonin content in pig blood provided by the present invention has simple implementation steps and good effects, and is also highly scalable. BRIEF DESCRIPTION OF THE DRAWINGS
[0061] Figure 1 The figures show the levels of melatonin in the peripheral blood of pigs when tryptophan in different doses was added for two days: A shows the changes of MT concentration in peripheral blood of the 0.1% feed ratio group at different stages (melatonin concentration in the first four days of feeding VS melatonin concentration in the first two days of feeding VS melatonin concentration in the second days after feeding); B shows the changes of MT concentration in peripheral blood of the 0.3% feed ratio group at different stages (melatonin concentration in the first four days of feeding VS melatonin concentration in the first two days of feeding VS melatonin concentration in the second days after feeding); C shows the changes of MT concentration in peripheral blood of the 0.5% feed ratio group at different stages (melatonin concentration in the first four days of feeding VS melatonin concentration in the first two days of feeding VS melatonin concentration in the second days after feeding); D shows the comparison of MT concentration in peripheral blood of different feed ratio groups (melatonin concentration in the 0.1% feed ratio group VS melatonin concentration in the 0.3% feed ratio group VS melatonin concentration in the 0.5% feed ratio group) after two days of feeding.
[0062] Figure 2 The figures show the levels of melatonin in the peripheral blood of pigs after four days of adding tryptophan at different doses: A shows the changes in peripheral blood MT concentration in the 0.1% feed ratio group at different stages (melatonin concentration in the first four days of feeding VS melatonin concentration in the first two days of feeding VS melatonin concentration in the last four days of feeding); B shows the changes in peripheral blood MT concentration in the 0.3% feed ratio group at different stages (melatonin concentration in the first four days of feeding VS melatonin concentration in the first two days of feeding VS melatonin concentration in the last four days of feeding); C shows the changes in peripheral blood MT concentration in the 0.5% feed ratio group at different stages (melatonin concentration in the first four days of feeding VS melatonin concentration in the first two days of feeding VS melatonin concentration in the last four days of feeding); D shows the comparison of peripheral blood MT concentrations in different feed ratio groups (melatonin concentration in the 0.1% feed ratio group VS melatonin concentration in the 0.3% feed ratio group VS melatonin concentration in the 0.5% feed ratio group) after four days of feeding. DETAILED DESCRIPTION
[0063] definition
[0064] Unless otherwise defined, all technical and scientific terms used in the present invention have the same meaning as commonly used in the field to which the present invention belongs. For the purpose of interpreting this specification, the following definitions will apply, and where appropriate, terms used in the singular will also include the plural form, and vice versa.
[0065] As used herein, the term "about" refers to a range of ±20% of the value that follows. In some embodiments, the term "about" refers to a range of ±10% of the value that follows. In some embodiments, the term "about" refers to a range of ±5% of the value that follows.
[0066] The present invention is further described in detail below through specific examples.
[0067] 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.
[0068] The experimental methods in the following examples without specifying specific conditions are usually carried out under conventional conditions or under conditions recommended by the manufacturer. The materials and reagents used in the examples are commercially available unless otherwise specified. The manufacturers of the reagents used are indicated, and similar products from other manufacturers are substitutes.
[0069] Experimental methods
[0070] 1. Tryptophan treatment plan
[0071] In the following examples, tryptophan was added according to the feed ratio of 0.1wt%, 0.3wt% and 0.5wt%, respectively, 5g, 15g and 25g; the feeding time of tryptophan was 7:00 to 8:00 in the morning every day; the feeding method was feed addition; the number of feedings per day was single.
[0072] 2. Blood Sample Collection
[0073] The blood was collected four days before tryptophan feeding, two days before tryptophan feeding, two days after tryptophan feeding, and four days after tryptophan feeding; the blood was collected from the anterior vena cava for analyzing the changes in melatonin.
[0074] 3. Determination of Melatonin Content
[0075] The following method for determining melatonin concentration is LC-MS detection (for specific detection methods, see references, Wang X, Liu T, Lv X, Sun N, Li F, Luo L, Zhuge X, Huang J, Wang LA Potential Nontraditional Approach To Combat tmexCD1-toprJ1-Mediated Tigecycline Resistance: Melatonins a Synergistic Adjuvant of Tigecycline. Antimicrob Agents Chemother. 2023Jul18; 67(7): e0004723. doi: 10.1128 / aac.00047-23. Epub 2023Jun 8. PMID: 37289048; PMCID: PMC10353380.); the basal level of melatonin in the peripheral blood of pigs was detected four days and two days before tryptophan feeding, and the changes in melatonin levels were detected two days and four days after tryptophan feeding.
[0076] Example 1
[0077] 1. Changes in peripheral blood melatonin levels after two days of adding different doses of tryptophan
[0078] Eighteen healthy gilts with similar body condition and age were selected, with a weight of about 100 kg. According to the addition ratio of tryptophan in the feed, they were randomly divided into 0.1% feed ratio group, 0.3% feed ratio group and 0.5% feed ratio group, with 6 pigs in each group.
[0079] Blood was collected from each group four days and two days before tryptophan feeding, and the basal level of melatonin was detected by LC-MS. Blood was collected two days after tryptophan feeding to detect melatonin levels. The results are shown in Figure 1 .
[0080] Figure 1In the figure, A shows the changes of MT concentration in peripheral blood in the 0.1% feed ratio group at different stages (melatonin concentration in the first four days of feeding VS melatonin concentration in the first two days of feeding VS melatonin concentration in the second days of feeding), which are as follows: (28.03±10.42VS26.18±4.517VS 57.74±6.832) n=6 (unit: head); B shows the changes of MT concentration in peripheral blood in the 0.3% feed ratio group at different stages (melatonin concentration in the first four days of feeding VS melatonin concentration in the first two days of feeding VS melatonin concentration in the second days of feeding), which are as follows: (17.70±6.614VS26.06±3.635VS 92.12±19.90)n=6 (unit: head); C is the change of MT concentration in peripheral blood in the 0.5% feed ratio group at different stages (melatonin concentration in the first four days of feeding VS melatonin concentration in the first two days of feeding VS melatonin concentration in the last two days of feeding), specifically as follows: (15.59±4.716VS27.12±3.022VS112.4±23.50)n=6 (unit: head); D is the comparison of MT concentration in peripheral blood in different feed ratio groups (melatonin concentration in 0.1% feed ratio group VS melatonin concentration in 0.3% feed ratio group VS melatonin concentration in 0.5% feed ratio group) after two days of feeding, specifically as follows: (57.74±6.832VS 92.12±19.90VS112.4±23.50)n=6 (unit: head). All data are expressed as mean ± standard error. Different letters indicate significant differences within the groups (p<0.05), and the same letters indicate no significant differences within the groups (p>0.05).
[0081] pass Figure 1 It can be seen that when the tryptophan addition ratio in the feed is 0.1%, 0.3% and 0.5%, the melatonin level in the peripheral blood of pigs can be significantly increased two days after tryptophan addition (p<0.05); when the melatonin levels of different addition ratio groups were compared two days after tryptophan addition, it was found that the melatonin level in the peripheral blood of the 0.5% tryptophan addition ratio group was significantly higher than that of the 0.1% tryptophan addition ratio group (p<0.05), and there was an increasing trend compared with the 0.3% tryptophan addition ratio group.
[0082] 2. Changes in peripheral blood melatonin levels after four days of adding different doses of tryptophan
[0083] Eighteen healthy gilts with similar body condition and age were selected and randomly divided into 0.1% feed ratio group, 0.3% feed ratio group and 0.5% feed ratio group according to the ratio of tryptophan added in feed, with 6 gilts in each group.
[0084] Blood was collected from each group four days and two days before tryptophan feeding, and the basal level of melatonin was detected by LC-MS. Blood was collected four days after tryptophan feeding to detect melatonin levels. The results are shown in Figure 2 ( Figure 2 The results after 4 days of feeding were essentially Figure 1 The results of the corresponding experiment were as follows: Figure 1 Continue feeding based on the results obtained, and then screen out the dosage and time that can stably increase melatonin).
[0085] Figure 2 In the figure, A shows the changes of MT concentration in peripheral blood in the 0.1% feed ratio group at different stages (melatonin concentration in the first four days of feeding VS melatonin concentration in the first two days of feeding VS melatonin concentration in the last four days of feeding), as follows: (28.03±10.42VS26.18±4.517VS 69.15±11.97) n=6 (unit: head); B shows the changes of MT concentration in peripheral blood in the 0.3% feed ratio group at different stages (melatonin concentration in the first four days of feeding VS melatonin concentration in the first two days of feeding VS melatonin concentration in the last four days of feeding), as follows: (17.70±6.614VS26.06±3.635VS 44.80±5.106)n=6 (unit: head); C is the change of MT concentration in peripheral blood in the 0.5% feed ratio group at different stages (melatonin concentration in the first four days of feeding VS melatonin concentration in the first two days of feeding VS melatonin concentration in the last four days of feeding), specifically as follows: (15.59±4.716VS27.12±3.022VS46.38±7.625)n=6 (unit: head); D is the comparison of MT concentration in peripheral blood in different feed ratio groups (melatonin concentration in 0.1% feed ratio group VS melatonin concentration in 0.3% feed ratio group VS melatonin concentration in 0.5% feed ratio group) after four days of feeding, specifically as follows: (69.15±11.97VS 44.80±5.106VS 46.38±7.625)n=6 (unit: head). All data are expressed as mean ± standard error. Different letters indicate significant differences within the groups (p<0.05), and the same letters indicate no significant differences within the groups (p>0.05).
[0086] pass Figure 2 It can be seen that when the tryptophan addition ratio in the feed is 0.1%, 0.3% and 0.5%, the melatonin level in the peripheral blood of pigs can be significantly increased four days after tryptophan addition (p<0.05); when the melatonin levels of different addition ratio groups were compared after four days of tryptophan addition, it was found that there was no significant difference in the melatonin level in the peripheral blood of pigs in the 0.1% tryptophan addition group, 0.3% tryptophan addition group and 0.5% tryptophan addition group (p>0.05).
[0087] Figure 1 and Figure 2 The results showed that the concentration of melatonin in the peripheral blood of pigs could be stably increased by adding tryptophan at a feed ratio of 0.1%.
[0088] The technical solution of the present invention is not limited to the above-mentioned specific embodiments. All technical variations made according to the technical solution of the present invention fall within the protection scope of the present invention.
Claims
1. A method for increasing the melatonin content in pig blood, comprising administering tryptophan or a salt thereof to the pig.
2. The method according to claim 1, characterized in that The administration of tryptophan or its salt includes oral, nasal, inhalation, parenteral, intravenous, intraperitoneal, subcutaneous, intramuscular, intradermal, topical or rectal administration; further oral administration.
3. The method according to any one of claims 1 to 2, characterized in that: The administration amount of tryptophan or its salt is 0.025-0.3 g / kg pig.day; further 0.04-0.26 g / kg pig.day; Preferably, the tryptophan or its salt is administered 1-3 times per day; Preferably, the tryptophan or its salt is administered for 1-6 days; further for 2-4 days.
4. The method according to any one of claims 1 to 3, characterized in that: The administration amount of the tryptophan or its salt is 0.24-0.26 g / kg pig day, and the administration days of the tryptophan or its salt are 1-3 days; or The administration amount of the tryptophan or its salt is 0.14-0.16 g / kg pig.day, and the administration days of the tryptophan or its salt are 1-3 days; or The administration amount of the tryptophan or its salt is 0.04-0.06 g / kg pig.day, and the administration days of the tryptophan or its salt are 1-5 days; Preferably, the tryptophan or its salt is administered every morning.
5. The method according to any one of claims 1 to 4, characterized in that: The tryptophan or its salt is added to feed and administered to pigs; Preferably, the pig is a female pig; Preferably, the step of increasing the melatonin content in pig blood is increasing the melatonin content in pig peripheral blood.
6. Use of tryptophan or its salts in a1) or a2): a1) Increase the melatonin content in pig blood; a2) preparing a product for increasing the melatonin content in pig blood.
7. The use according to claim 6, characterized in that: The product further comprises: an instruction manual, wherein the instruction manual records: the method for increasing the melatonin content in pig blood as described in any one of claims 1-4.
8. The use according to any one of claims 6 to 7, characterized in that: The product also contains other active ingredients for increasing the melatonin content in pig blood.
9. The use according to any one of claims 6 to 8, characterized in that: The product is a feed additive or a feed.
10. The method or use according to any one of claims 1 to 9, characterized in that: The salt includes at least one of a metal salt, an ammonium salt, a salt formed with an organic base, a salt formed with an inorganic acid, a salt formed with an organic acid, a salt formed with a basic amino acid, and a salt formed with an acidic amino acid.