Use of component A as or for preparing SIRT5 inducer

Abstract

The invention provides an application of a component A as or used for preparing an SIRT5 inducer, the component A at least comprises one or more than two of ergothioneine, ectoine and hyaluronic acid or salt thereof, and the component A can be used for the SIRT5 inducer or used for preparing an SIRT5 expression inducer, so that cell aging can be inhibited, and the SIRT5 inducer can be used for preparing an SIRT5 expression inducer. The traditional Chinese medicine composition is used for improving natural skin aging, cardiac hypertrophy and / or hyperammonemia.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of raw material efficacy research, and particularly relates to the use of component A as or for preparing a SIRT5 inducer. BACKGROUND

[0002] Cell aging (also known as cellular senescence) is a process in which cells gradually lose their ability to divide and eventually enter a permanent growth-arrested state. This process is closely related to the occurrence and development of various diseases. For example, the impaired function of aging endothelial cells can lead to vascular sclerosis, elevated blood pressure, and myocardial hypertrophy and myocardial fibrosis, and the accumulation of aging cells can also affect the repair and regeneration capacity of blood vessels; the aging of brain cells is related to neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease; the aging of skeletal cells can lead to decreased bone mass and osteoporosis, increasing the risk of bone fracture; the aging of skin cells can be accompanied by problems such as fibrotic diseases similar to scleroderma, skin laxity, wrinkles, and wound healing disorders; the aging of liver cells leads to decreased liver cell function, reducing the liver's ability to metabolize ammonia, leading to hyperammonemia, which can further lead to cirrhosis and liver fibrosis; in addition, cell aging can lead to insulin resistance and decreased function of pancreatic beta cells, thereby increasing the risk of type 2 diabetes.

[0003] Fibroblasts are the most common cell type in connective tissue, which exist in parts such as blood vessel walls, bones, skin, and joints. The aging of fibroblasts has multiple effects on the body, for example, the aging of fibroblasts can lead to thinning of the blood vessel wall, decreased elasticity, myocardial hypertrophy, cardiac fibrosis, and increased risk of cardiovascular disease, and can also lead to slow wound healing in the skin, low repair efficiency after tissue damage, and even abnormal structure and function of tissues, such as vascular sclerosis, liver cell fibrosis, cirrhosis, hyperammonemia, decreased bone density, and joint diseases such as joint stiffness. It can be seen that the aging of fibroblasts has a profound impact on multiple systems and organs of the body, easily leading to decreased tissue function and increasing the risk of disease in multiple organs.

[0004] Sirtuins (silent information regulator-related enzymes) are a class of NAD (nicotinic acid adenine dinucleotide) dependent deacetylases that play an important role in the regulation of cell aging. In the human body, there are various subtypes of Sirtuins such as SIRT1-SIRT7, each with different focuses and action scenarios.

[0005] SIRT1 is mainly located in the nucleus, SIRT1 can activate some anti-aging related genes by deacetylation, inhibit the expression of aging related genes, and is considered as a long life related protein. SIRT1 is involved in the regulation of a series of wide physiological processes, including inflammation, apoptosis, cell aging, longevity, glucose homeostasis, aging, etc. The solvent extract of the plant body of Peucedanum japonicum Thunb. provided in Chinese patent CN113768959B can activate Sirtuin-1 and inhibit cell aging caused by oxidative stress.

[0006] SIRT2 is mainly located in the cytoplasm, has mono-ADP-ribosyltransferase and deacetylase activity, is involved in cell cycle regulation and maintenance of microtubule stability, and ensures the integrity of cell structure. SIRT2 is overexpressed in neurodegenerative diseases, protects neural cells from oxidative damage caused by aging and disease, and its abnormal expression is related to cancer (glioma, melanoma). SIRT2 plays a crucial role in CNS function by delivering from oligodendrocytes to neurons through exosomes. SIRT2 is related to neuropsychiatric diseases, and clinical studies have shown that SIRT2 transcripts are reduced in peripheral leukocytes and hippocampal tissue of patients with depression. In rodents, hippocampal SIRT2 reduction is associated with the progression of depression and impaired neural plasticity, which can be a new therapeutic target for preventing depression (Rui Liu et. 2015, SIRT2 is involved in the modulation of depressive behaviors).

[0007] SIRT3, SIRT4 and SIRT5 are mainly located in mitochondria, and they may affect the process of cell aging by participating in the regulation of mitochondrial metabolism and stress response. Sirtuins are considered as potential targets for delaying cell aging and treating cell aging related diseases. By activating Sirtuins or developing their activators, it may help to slow down the process of cell aging and improve the disease state related to cell aging. For example, the level of SIRT5 protein in the heart is high, and SIRT5 gene knockout mice also develop hypertrophic cardiomyopathy. And knocking out SIRT5 causes mice to develop hyperammonemia and liver damage (Melanie Gertzet. Function and regulation of the mitochondrial Sirtuin isoform Sirt5 in Mammalia, 2010). Cardiac hypertrophy is an adaptive response of cardiomyocytes to pressure load (such as hypertension, aortic stenosis), manifested as cardiomyocyte hypertrophy, fibrosis and metabolic disorder. If it continues to progress, it can lead to heart failure, arrhythmia and even sudden death.

[0008] In summary, SIRTs have their own characteristics in the regulatory pathways, and they play roles in different biological processes and are associated with various diseases. SIRT5 regulates the succinylation and glutarylation status of metabolic enzymes in mitochondria. So far, there are still few activators for regulating the activity of SIRT5, but SIRT5 also plays an important role in the human body, so it is still necessary to develop more activators for SIRT5.

[0009] Ergothioneine (EGT), also known as mercosylhistidine betaine, is a natural antioxidant. In the human body, ergothioneine can be transported to mitochondria by the transporter OCTN-1 in cells, directly scavenge active oxygen free radicals, and play the role of antioxidant and cell protection. However, there is no relevant report on the influence of ergothioneine on the expression of SIRT5 and the improvement of myocardial hypertrophy and / or hyperammonemia.

[0010] Hyaluronan (HA) is a glycosaminoglycan widely present in the extracellular matrix, which is composed of a disaccharide repeat sequence of N-acetylglucosamine and glucuronic acid. Hyaluronan is widely used in the fields of medicine, food, cosmetics, etc. However, there is no relevant report on the influence of hyaluronan oligosaccharide on the expression of SIRT5 and myocardial hypertrophy and / or hyperammonemia.

[0011] Ectoine belongs to cyclic amino acids, and is a low-molecular-weight organic compound naturally produced in many bacteria and some extremophilic organisms. The unique cyclic structure of ectoine endows it with high stability and biological activity. In recent years, ectoine has attracted widespread attention due to its excellent moisturizing, antioxidant and anti-stress effects. However, there is no relevant report on the influence of ectoine on the expression of SIRT5 and myocardial hypertrophy and / or hyperammonemia. SUMMARY

[0012] The inventors of the present application accidentally found that ergothioneine, ectoine and / or hyaluronic acid or a salt thereof have the effect of increasing the expression amount of SIRT5, thereby completing the present application.

[0013] The specific technical solutions of the present application are as follows: 1. Use of component A as or for preparing a SIRT5 inducer, wherein the component A at least contains one or more than two of ergothioneine, ectoine and hyaluronic acid or a salt thereof.

[0014] 2. Use of component A in preparing a product for increasing the expression amount of SIRT5, wherein the component A at least contains one or more than two of ergothioneine, ectoine and hyaluronic acid or a salt thereof.

[0015] 3. The use according to item 1 or 2, wherein the component A is used for the preparation of an anti-cellular aging preparation.

[0016] 4. The use according to item 1 or 2, the component A is used for the improvement of cardiac hypertrophy.

[0017] 5. The use according to item 1 or 2, the component A is used for the improvement of hyperammonemia.

[0018] 6. The use of the component A for the preparation of a preparation for the improvement of cardiac hypertrophy, wherein the component A comprises one or two or more of ergothioneine, icotidine and hyaluronic acid or a salt thereof.

[0019] 7. The use of the component A for the preparation of a preparation for the improvement of hyperammonemia, wherein the component A comprises one or two or more of ergothioneine, icotidine and hyaluronic acid or a salt thereof.

[0020] Effects of the Invention The component A comprising one or two or more of ergothioneine, icotidine and hyaluronic acid or a salt thereof can increase the expression amount of SIRT5, i.e. the component A can be used as a SIRT5 inducer or for the preparation of an inducer of SIRT5 expression, thereby it can inhibit cellular aging, be used for the improvement of natural skin aging, cardiac hypertrophy and / or hyperammonemia. BRIEF DESCRIPTION OF DRAWINGS

[0021] Figure 1 Figure 1 is a fluorescence graph of the effect of ergothioneine on the expression of SIRT2, SIRT5 protein in human skin fibroblasts in Example 1.

[0022] Figure 2 Figure 3 is a fluorescence graph of the effect of hyaluronic acid or a salt thereof on the expression of SIRT2, SIRT5 protein in human skin fibroblasts in Example 1.

[0023] Figure 3 Figure 4 is a fluorescence graph of the effect of icotidine on the expression of SIRT2, SIRT5 protein in human skin fibroblasts in Example 1. DETAILED DESCRIPTION

[0024] The present application is described in detail below with reference to the described embodiments. Although specific embodiments of the present application are shown, it is understood that the present application can be implemented in various forms and should not be limited by the embodiments set forth herein. Rather, these embodiments are provided so that the present application can be more thoroughly understood and the scope of the present application can be accurately conveyed to those skilled in the art.

[0025] It should be noted that some terms are used in the description and claims herein. Those skilled in the art will understand that one name can be used instead of another to refer to the same component. The description and claims herein do not distinguish components based on the difference in names, but rather on the difference in functions. As mentioned throughout the description and claims, "comprising" or "including" is an open term, which should be interpreted as "including but not limited to". The subsequent description is a preferred embodiment of the present application, but the description is for the purpose of illustrating the general principles of the description, and is not intended to limit the scope of the present application. The scope of protection of the present application is defined by the appended claims.

[0026] The present application provides a use of component A for or in the preparation of a SIRT5 inducer, wherein the component A comprises at least one or two or more of ergothioneine, icodextrin and hyaluronic acid or a salt thereof.

[0027] The present application provides a use for increasing the expression of SIRT5, wherein the component A comprises at least one or two or more of ergothioneine, icodextrin and hyaluronic acid or a salt thereof.

[0028] The inventors of the present application have unexpectedly found that a component A comprising at least one or two or more of ergothioneine, icodextrin and hyaluronic acid or a salt thereof can be used to increase the expression of SIRT5 protein, and thus can be used for or in the preparation of a SIRT5 inducer.

[0029] In the present application, the use concentration of component A is not limited, and those skilled in the art can make routine selection based on actual needs, for example, the concentration of component A can be 0.001-2 mg / mL.

[0030] For example, the concentration of the component A can be 0.001 mg / mL, 0.002 mg / mL, 0.003 mg / mL, 0.004 mg / mL, 0.005 mg / mL, 0.006 mg / mL, 0.007 mg / mL, 0.008 mg / mL, 0.009 mg / mL, 0.01 mg / mL, 0.02 mg / mL, 0.03 mg / mL, 0.04 mg / mL, 0.05 mg / mL, 0.06 mg / mL, 0.07 mg / mL, 0.08 mg / mL, 0.09 mg / mL, 0.1 mg / mL, 0.2 mg / mL, 0.3 mg / mL, 0.4 mg / mL, 0.5 mg / mL, 0.6 mg / mL, 0.7 mg / mL, 0.8 mg / mL, 0.9 mg / mL, 1.0 mg / mL, 1.1 mg / mL, 1.2 mg / mL, 1.3 mg / mL, 1.4 mg / mL, 1.5 mg / mL, 1.6 mg / mL, 1.7 mg / mL, 1.8 mg / mL, 1.9 mg / mL, 2.0 mg / mL, and the like.

[0031] In some embodiments, the hyaluronic acid or salt thereof has a molecular weight of 400-5000 Da.

[0032] For example, the hyaluronic acid or salt thereof can have a molecular weight of 400 Da, 500 Da, 600 Da, 700 Da, 800 Da, 900 Da, 1000 Da, 1100 Da, 1200 Da, 1300 Da, 1400 Da, 1500 Da, 1600 Da, 1700 Da, 1800 Da, 1900 Da, 2000 Da, 2100 Da, 2200 Da, 2300 Da, 2400 Da, 2500 Da, 2600 Da, 2700 Da, 2800 Da, 2900 Da, 3000 Da, 3100 Da, 3200 Da, 3300 Da, 3400 Da, 3500 Da, 3600 Da, 3700 Da, 3800 Da, 3900 Da, 4000 Da, 4100 Da, 4200 Da, 4300 Da, 4400 Da, 4500 Da, 4600 Da, 4700 Da, 4800 Da, 4900 Da, 5000 Da, and any molecular weight therebetween, and the like.

[0033] In the present application, the method for determining the molecular weight of hyaluronic acid or its salt is not limited, and the skilled person in the art can determine it by using the conventional method in the art, for example, by using the conventional gel permeation chromatography (GPC), viscosity method, laser light scattering method.

[0034] In some embodiments, the component A is used for improving natural skin aging, myocardial hypertrophy and / or hyperammonemia.

[0035] As described above, the component A can increase the expression amount of SIRT5 protein, thereby inhibiting cell aging or reversing aging cells, and can be used for treating related diseases caused by cell aging, such as natural skin aging, myocardial hypertrophy and / or hyperammonemia.

[0036] The present application provides the use of the component A in the preparation of a product for improving natural skin aging, myocardial hypertrophy and / or hyperammonemia, wherein the component A at least comprises one or two or more of ergothioneine, icodextrin and hyaluronic acid or its salt. In some embodiments, the molecular weight of the hyaluronic acid or its salt is 400-5000 Da.

[0037] In the present application, the amount of the component A used is not limited, and the skilled person in the art can make a conventional selection based on actual needs, for example, the concentration of the component A can be 0.001-2 mg / mL.

[0038] The present application provides a composition for use as or in the preparation of a SIRT5 inducer, which at least comprises one or two or more of ergothioneine, icodextrin and hyaluronic acid or its salt. The present application also provides a composition for improving natural skin aging, myocardial hypertrophy and / or hyperammonemia, which at least comprises one or two or more of ergothioneine, icodextrin and hyaluronic acid or its salt.

[0039] In the present application, the mass percentage of the component A in the composition is not limited, and the skilled person in the art can make a conventional selection based on actual needs. For example, the content of the component A can be 0.001-2 mg / mL in terms of the mass percentage in the composition.

[0040] In some embodiments, the molecular weight of the hyaluronic acid or its salt is 400-5000 Da.

[0041] For example, the molecular weight of the hyaluronic acid or its salt can be 400 Da, 500 Da, 600 Da, 700 Da, 800 Da, 900 Da, 1000 Da, 1100 Da, 1200 Da, 1300 Da, 1400 Da, 1500 Da, 1600 Da, 1700 Da, 1800 Da, 1900 Da, 2000 Da, 2100 Da Da, 2200 Da, 2300 Da, 2400 Da, 2500 Da, 2600 Da, 2700 Da, 2800 Da, 2900 Da, 3000 Da, 3100 Da, 3200 Da, 3300 Da, 3400 Da, 3500 Da, 3600 Da, 3700 Da, 3800 Da, 3900 Da, 4000 Da Da, 4100 Da, 4200 Da, 4300 Da, 4400 Da, 4500 Da, 4600 Da, 4700 Da, 4800 Da, 4900 Da, 5000 Da, etc.

[0042] Example This application provides a general and / or specific description of the materials and test methods used in the experiments. In the following examples, unless otherwise specified, % represents wt%, i.e., weight percentage. Reagents or instruments used, unless otherwise specified, are all commercially available conventional reagent products.

[0043] Example 1: Assay on the regulation of SIRT2 and SIRT5 protein expression in human skin fibroblasts Immunofluorescence assays were used to determine the relative levels of the SIRT protein family (SIRT2 and SIRT5) in human skin fibroblasts under UVA irradiation using semi-quantitative techniques. This method can be used to assess the effect of the test sample on the expression of SIRT protein family proteins.

[0044] 1. Reagents, Consumables and Instruments Main reagents: Bioyouth™-EGT Pure ultrapure ergothioneine was purchased from Bloomage Biotechnology Co., Ltd.; Hybloom TM Micro-genuine sodium hyaluronate (molecular weight 800-1000 Da) was purchased from Bloomage Biotechnology Co., Ltd.; Xi'an Yan TMEctoine was purchased from Huaxi Biotechnology Co., Ltd.; fetal bovine serum, DMEM medium, antibiotics, PBS were purchased from Thermo Fisher Scientific, USA; SIRT2, SIRT5 antibody were purchased from Abeam, UK; Anti-Fade Mounting Medium with DAPI was purchased from Vectashield, USA; Bovine serum albumin (BSA) was purchased from Sigma, USA; Anhydrous methanol was purchased from National Pharmaceutical Group Chemical Reagent Co., Ltd.

[0045] Main instruments: microscope, fluorescence microscope were purchased from Leica, Germany.

[0046] Natural aging human skin fibroblasts: normal human skin fibroblasts were purchased from ATCC (catalog number: PCS-201-010), which were obtained by continuous subculture of normal human fibroblasts in vitro. When the cells were subcultured to P30 to P35 or so, the natural aging fibroblasts were obtained by detecting the cells by the conventional method in the art.

[0047] 2. Test method 1) Cell inoculation: natural aging human skin fibroblasts were taken and cultured with DMEM medium containing 10% FBS. When the cell density reached about 80%, the cells were digested with 0.05% trypsin, centrifuged at 1000 rpm for 5 min, resuspended and counted, inoculated in 24-well plates, and cultured at 37°C, 5% CO2.

[0048] 2) Sample treatment: after 24 h of inoculation, the culture medium was removed, and the test group was added with corresponding concentration samples, and the blank group and the control group were added with DMEM complete culture medium. Continue to incubate in the incubator for 72 h. After the exposure is completed, the above samples are discarded, washed with PBS, and then fixed with cold methanol.

[0049] 3) Staining: the cell climbing sheet was taken out, blocked with BSA for 30 min, incubated with the corresponding antibody overnight, discarded the primary antibody, washed with PBS, added with the corresponding secondary antibody and DAPI, and incubated in the dark, finally added with the mounting agent, and photographed under the same exposure condition using fluorescence microscope, and the relative average fluorescence intensity of the related protein in the cells was analyzed using software.

[0050] 3. Statistical analysis In this example, the data of three repeated experiments were represented as Mean ± SD, and t test was used for comparison with the control group, P <0.05 indicates a statistically significant difference, P <0.01 indicates a significant statistically significant difference, P <0.001 is a very significant statistically significant difference.

[0051] 4. Judgment criteria If the relative mean fluorescence intensity of the sample group (%) is greater than 100%, and the result has a statistical difference compared with the control group (P<0.05), it is determined that the sample can improve the relative mean fluorescence intensity of the relevant protein at this concentration. P <0.05), it is determined that the sample can improve the relative mean fluorescence intensity of the relevant protein at this concentration.

[0052] 5. Test results (1) The sample ergothioneine has an up-regulating effect on the expression of SIRT2 and SIRT5 proteins in naturally aged human fibroblasts, and the test results are shown in Table 1 and Figure 1 .

[0053] As can be seen from Table 1 and Figure 1 , compared with the control group, the relative fluorescence intensity of SIRT5 protein after ergothioneine treatment is 110±6%, P<0.01, with a very significant statistical difference; the relative fluorescence intensity of SIRT2 protein in the ergothioneine group is 102±5%, P>0.05, without a statistical difference. Therefore, ergothioneine has an effect of up-regulating the relative fluorescence intensity of SIRT5 protein, but does not have an effect of up-regulating the relative fluorescence intensity of SIRT2 protein. P P

[0054] As described above, the expression of SIRT5 protein in fibroblasts caused by natural aging can be improved, so that skin natural aging, myocardial hypertrophy and / or hyperammonemia can be improved.

[0055] Table 1 Effect of ergothioneine on the relative fluorescence intensity of SIRT protein in naturally aged human skin fibroblasts

[0056] (2) Sodium hyaluronate has an up-regulating effect on the expression of SIRT2 and SIRT5 proteins in naturally aged human skin fibroblasts, and the test results are shown in Table 2 and Figure 2 .

[0057] As can be seen from Table 2 and Figure 2 , compared with the blank group, the relative fluorescence intensity of SIRT5 protein after sodium hyaluronate treatment is 108%, P<0.01, with a very significant statistical difference; the relative fluorescence intensity of SIRT2 protein in the sodium hyaluronate treatment group is 99±5%, P>0.05, without a statistical difference. Therefore, sodium hyaluronate has an effect of up-regulating the relative fluorescence intensity of SIRT5 protein, but does not have an effect of up-regulating the relative fluorescence intensity of SIRT2 protein. P P

[0058] As described above, sodium hyaluronate can improve the expression of SIRT5 protein in fibroblasts caused by natural aging, so that it can be used to improve skin natural aging, myocardial hypertrophy and / or hyperammonemia.​​​​

[0059] Table 2 Effect of sodium hyaluronate on the relative fluorescence intensity of SIRT protein in naturally aged human fibroblasts

[0060] (3) Ecodin has an up-regulating effect on the expression of SIRT2 and SIRT5 proteins in naturally aged human skin fibroblasts, and the test results are shown in Table 3 and Figure 3 .

[0061] As can be seen from Table 3 and Figure 3 , compared with the blank group, the relative fluorescence intensity of SIRT5 protein after ecodin treatment was 109±4%, P <0.001, with extremely significant statistical difference; the relative fluorescence intensity of SIRT2 protein in the ecodin group was 104±5%, P >0.05, with no statistical difference. Therefore, ecodin has an effect of up-regulating the relative fluorescence intensity of SIRT5 protein, and it does not have an effect of up-regulating the relative fluorescence intensity of SIRT2 protein.

[0062] According to the above experimental results, ecodin can improve the expression of SIRT5 protein in fibroblasts caused by natural aging, and thus can be used to improve skin natural aging, myocardial hypertrophy and / or hyperammonemia.

[0063] Table 3 Effect of ecodin on the relative fluorescence intensity of SIRT protein in naturally aged human skin fibroblasts

[0064] In summary, ergothioneine, ecodin and / or hyaluronic acid or its salt can up-regulate the expression of SIRT5 protein in naturally aged human skin fibroblasts, and therefore one or more of ergothioneine, ecodin and hyaluronic acid or its salt has the potential to be and be used as a SIRT5 inducer.

[0065] The above is only a preferred embodiment of the present application, and does not limit the present application in other forms. Any person skilled in the art can modify or modify the above disclosed technical content into equivalent embodiments with equivalent changes. However, any simple modification, equivalent change and modification made to the above embodiments without departing from the technical solution of the present application, and according to the technical essence of the present application, still belongs to the protection scope of the technical solution of the present application.

Claims

1. Use of component A in the preparation of a SIRT5 inducer, wherein component A comprises at least one or more of ergothioneine, ectoine, and hyaluronic acid or salts thereof.

2. Use of component A in the preparation of a product that enhances SIRT5 expression, wherein component A comprises at least one or more of ergothioneine, ectoine, and hyaluronic acid or salts thereof.

3. The use according to claim 1 or 2, wherein component A is used to prepare an anti-cellular aging product.

4. The use according to claim 1 or 2, wherein component A is used to improve myocardial hypertrophy.

5. The use according to claim 1 or 2, wherein component A is used to improve hyperammonemia.

6. Use of component A in the preparation of articles for improving myocardial hypertrophy, wherein component A comprises one or more of ergothioneine, ectoine and hyaluronic acid or salts thereof.

7. Use of component A in the preparation of an article for improving hyperammonemia, wherein component A comprises one or more of ergothioneine, ectoine and hyaluronic acid or salts thereof.

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