Application of Cyp2c29 gene in preparation of medicine for delaying senescence and / or improving cognitive impairment

Abstract

The invention belongs to the technical field of molecular diagnosis, and particularly relates to application of a Cyp2c29 gene in preparation of a medicine for delaying senescence and / or improving cognitive impairment, and a marker comprises a CYP2C family gene and an expression product thereof and / or a Cyp2c family gene and an expression product thereof. The marker comprises a CYP2C family gene and an expression product thereof and / or a Cyp2c family gene and an expression product thereof, the marker has high specificity and sensitivity, and by using the marker as a detection index, the aging degree and the cognitive impairment risk of an individual can be rapidly and accurately evaluated, and the detection result is accurate. Scientific basis is provided for early intervention, clinical diagnosis or personalized medical treatment of individual senescence and / or cognitive impairment, and the application prospect is wide.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of molecular diagnosis, and particularly relates to application of Cyp2c29 gene in preparation of a drug for delaying aging and / or improving cognitive impairment. BACKGROUND

[0002] According to the latest world population data released by the United Nations, in 2024, the global population aged 65 and above accounted for 12% of the total population, and it is expected that this proportion will reach 20% by 2050, which reflects the accelerated development of global population aging. With the increase in age, the organs and systems of the human body, such as the brain, undergo degenerative changes, and the number of neurons and nerve cells decreases significantly, leading to a decline in learning ability, working memory and thinking function, while the neurotransmitters and their receptors in the brain change, interfering with neural signal transmission and further affecting cognitive function.

[0003] At present, the detection of aging and / or cognitive impairment mainly relies on traditional biomarkers (such as biochemical indicators in blood), neuroimaging examination (such as MRI, PET) and gene detection technology (such as qPCR, Western Blot, etc.). However, these methods have many limitations, for example, traditional biomarkers have low specificity and sensitivity, and it is difficult to accurately reflect early changes; while neuroimaging examination equipment is expensive and complex to operate, and is not suitable for large-scale screening. In view of this, it is particularly important to explore a new high-specificity marker for detecting aging and / or cognitive impairment.

[0004] In recent years, the role of CYP2C family genes and their expression products in various diseases has gradually attracted attention. The enzymes encoded by CYP2C family genes play an important role in drug metabolism, endogenous substance metabolism and inflammatory response, and changes in their expression levels are closely related to various diseases (such as cardiovascular disease, diabetes, neurodegenerative disease, etc.).

[0005] CYP2C family genes are a subfamily of the human cytochrome P450 (Cytochrome P450) superfamily, and the enzymes encoded by this part of genes play an important role in the liver and other tissues, mainly participating in the metabolism of endogenous and exogenous substances, including drugs, hormones, environmental toxins, etc. At the same time, in other animals, homologous genes of CYP2C family genes, such as Cyp2c family genes, also play similar functions.

[0006] Therefore, it is of great significance to explore a new high-specificity marker for detecting aging and / or cognitive impairment and develop new uses of CYP2C family genes / Cyp2c family genes. SUMMARY

[0007] The present application aims to at least partially solve at least one of the technical problems existing in the prior art. To this end, the present application provides an application of a Cyp2c29 gene in the preparation of a drug for delaying aging and / or improving cognitive impairment. The marker of the present application includes a CYP2C family gene and its expression product and / or a Cyp2c family gene and its expression product, which has high specificity and sensitivity. By using the marker of the present application as a detection index, the aging degree and cognitive impairment risk of an individual can be quickly and accurately evaluated, which provides a scientific basis for early intervention, clinical diagnosis or personalized medicine of individual aging and / or cognitive impairment, and has a wide application prospect.

[0008] In a first aspect of the present application, a marker for detecting aging and / or cognitive impairment is provided. According to an embodiment of the present application, the marker includes a CYP2C family gene and its expression product and / or a Cyp2c family gene and its expression product. The marker of the present application includes a CYP2C family gene and its expression product and / or a Cyp2c family gene and its expression product, which has high specificity and sensitivity. By using the marker of the present application as a detection index, the aging degree and cognitive impairment risk of an individual can be quickly and accurately evaluated, which provides a scientific basis for early intervention, clinical diagnosis or personalized medicine of individual aging and / or cognitive impairment, and has a wide application prospect.

[0009] According to an embodiment of the present application, the above-mentioned marker can have the following additional technical features:

[0010] According to an embodiment of the present application, the CYP2C family gene includes one or more of CYP2C8, CYP2C9, CYP2C18 and CYP2C19.

[0011] According to an embodiment of the present application, the Cyp2c family gene includes one or more of Cyp2c29, Cyp2c38, Cyp2c37, Cyp2c40, Cyp2c50 and Cyp2c54.

[0012] According to a preferred embodiment of the present application, the CYP2C family gene is CYP2C9 and CYP2C19.

[0013] According to a preferred embodiment of the present application, the Cyp2c family gene is Cyp2c29.

[0014] In a second aspect, the present application provides a use of CYP2C family genes and expression products thereof and / or Cyp2c family genes and expression products thereof, and / or a reagent for detecting CYP2C family genes and expression products thereof and / or Cyp2c family genes and expression products thereof in the preparation of a kit. According to embodiments of the present application, the kit is used for detecting aging and / or cognitive impairment.

[0015] It is understood by those skilled in the art that the features and advantages described above for the marker for detecting aging and / or cognitive impairment also apply to the use, which will not be repeated here.

[0016] According to embodiments of the present application, the above-mentioned use can further have the following additional technical features:

[0017] According to embodiments of the present application, a decrease in the level of CYP2C family genes and expression products thereof and / or Cyp2c family genes and expression products thereof is indicative of aging and / or cognitive impairment.

[0018] According to embodiments of the present application, the CYP2C family genes include one or more of CYP2C8, CYP2C9, CYP2C18 and CYP2C19.

[0019] According to embodiments of the present application, the Cyp2c family genes include one or more of Cyp2c29, Cyp2c38, Cyp2c37, Cyp2c40, Cyp2c50 and Cyp2c54.

[0020] According to preferred embodiments of the present application, the CYP2C family genes are CYP2C9 and CYP2C19.

[0021] According to preferred embodiments of the present application, the Cyp2c family genes are Cyp2c29.

[0022] In a third aspect, the present application provides a kit for detecting aging and / or cognitive impairment. According to embodiments of the present application, the kit contains a reagent for detecting CYP2C family genes and expression products thereof and / or Cyp2c family genes and expression products thereof. According to embodiments of the present application, the kit can detect the expression level of CYP2C family genes and expression products thereof and / or Cyp2c family genes and expression products thereof. Further, the use of the kit of the present application for detecting a sample to be tested can quickly and accurately assess the degree of aging and the risk of cognitive impairment of an individual, and has a wide application prospect.

[0023] According to embodiments of the present application, the above-mentioned kit can further have the following additional technical features:

[0024] According to an embodiment of the present application, the CYP2C family gene comprises one or more of CYP2C8, CYP2C9, CYP2C18 and CYP2C19.

[0025] According to an embodiment of the present application, the Cyp2c family gene comprises one or more of Cyp2c29, Cyp2c38, Cyp2c37, Cyp2c40, Cyp2c50 and Cyp2c54.

[0026] According to a preferred embodiment of the present application, the CYP2C family gene is CYP2C9 and CYP2C19.

[0027] According to a preferred embodiment of the present application, the Cyp2c family gene is Cyp2c29.

[0028] In a fourth aspect of the present application, a method for constructing an animal model is provided. According to an embodiment of the present application, the method comprises the following steps: editing the genome of a target animal using genetic engineering techniques to modify a Cyp2c family gene, thereby obtaining a target animal model. According to the method of the present application, researchers can construct animal models with Cyp2c family gene silencing or the like, so as to further study the mechanism of Cyp2c family genes in aging and cognitive impairment, and further explore the potential functions and application values of homologous genes (CYP2C family genes) of Cyp2c family genes in human aging and cognitive impairment, and provide an experimental basis for early intervention, personalized medicine or new treatment strategy development for human individual aging and / or cognitive impairment.

[0029] According to an embodiment of the present application, the method for constructing an animal model can have the following additional technical features:

[0030] According to an embodiment of the present application, the Cyp2c family gene comprises one or more of Cyp2c29, Cyp2c38, Cyp2c37, Cyp2c40, Cyp2c50 and Cyp2c54.

[0031] According to an embodiment of the present application, the modification comprises one or more of knockdown, knockout, overexpression, gene silencing and gene activation.

[0032] According to an embodiment of the present application, the target animal comprises one or more of mice, rats, rabbits, pigs and non-human primates.

[0033] In a fifth aspect, the present application provides use of the CYP2C family gene and / or the Cyp2c family gene, the marker of the first aspect and / or the kit of the third aspect in the development or preparation of a drug for preventing, alleviating, assisting treatment or treating cognitive dysfunction.

[0034] It is understood by those skilled in the art that the features and advantages described above for the marker or the kit also apply to the use, which will not be described here again.

[0035] According to the embodiments of the present application, the above-mentioned use can have the following additional technical features:

[0036] According to the embodiments of the present application, the CYP2C family gene comprises one or more of CYP2C8, CYP2C9, CYP2C18 and CYP2C19.

[0037] According to the embodiments of the present application, the Cyp2c family gene comprises one or more of Cyp2c29, Cyp2c38, Cyp2c37, Cyp2c40, Cyp2c50 and Cyp2c54.

[0038] Additional aspects and advantages of the present application will be in part apparent and in part pointed out below in the description of embodiments. BRIEF DESCRIPTION OF DRAWINGS

[0039] The above and / or additional aspects and advantages of the present application will become apparent and be readily appreciated from the following description of embodiments, taken in conjunction with the accompanying drawings, in which:

[0040] Figure 1 Fig. 1 is a natural aging mouse lipid metabolism sequencing analysis and gene expression analysis result chart in the embodiment 1 of the present application, wherein A is a natural aging mouse experiment flow chart, B is a mouse liver lipid metabolism product sequencing analysis result chart, C is a COX, LOX and CYP metabolite proportion pie chart, and D is a natural aging mouse Cyp2c related gene mRNA expression level detection result chart;

[0041] Figure 2 Fig. 2 is a chemical induction aging mouse Cyp2c related gene mRNA expression analysis result chart in the embodiment 2 of the present application, wherein A is a D-Gal induced WT aging mouse experiment flow chart, and B is a D-Gal induced WT aging mouse Cyp2c related gene mRNA expression level detection result chart;

[0042] Figure 3Figures for the results of the grip strength ability and behavioral tests of the WT mice and Cyp2c29 KO mice in Example 3 of the present application, wherein A is a flow chart of the experiment of the D-Gal induced Cyp2c29 KO aging mice, B is a graph of the results of the grip strength ability test, C is a track graph of the open field test, D is a graph of the results of the total distance test of the open field test, E is a graph of the results of the rapid movement time test, F is a graph of the results of the novel object recognition index test, G is a graph of the results of the Y maze autonomous exploration times test, and H is a graph of the results of the Y maze time to enter the new arm test. DETAILED DESCRIPTION

[0043] Embodiments of the present application are described in detail below. The embodiments described below are examples only and are not intended to limit the present application, which can be embodied in various ways.

[0044] It should be noted that the terms "first", "second" are used only for descriptive purposes and should not be construed as indicating or implying relative importance or a specific order of the technical features indicated. Thus, features defined with "first", "second" can include one or more of the features implicitly or explicitly. Further, in the description of the present application, the meaning of "a plurality of" is two or more, unless otherwise specified.

[0045] The endpoints of the ranges and any values disclosed herein are not limited to the precise values stated. The ranges or values should be construed to be roughly about the ranges or values. For ranges, the endpoints are included as well as intervening points which are included in the ranges. For values, the values are included as well as intervening values which are included in the values.

[0046] In this document, the terms "comprises" or "comprising" are open-ended, that is, they mean including, but not limited to, the indicated features.

[0047] In this document, the terms "optionally", "optional" or "optional" generally mean that the subsequently described event or circumstance can or can not occur, and the description includes instances where the event or circumstance occurs and instances where it does not.

[0048] Marker for detecting aging and / or cognitive impairment

[0049] The present application provides a marker for detecting aging and / or cognitive impairment. According to an embodiment of the present application, the marker comprises a CYP2C family gene and an expression product thereof and / or a Cyp2c family gene and an expression product thereof. The marker of the present application comprises a CYP2C family gene and an expression product thereof and / or a Cyp2c family gene and an expression product thereof, which has high specificity and sensitivity, and by using the marker of the present application as a detection index, the aging degree and cognitive impairment risk of an individual can be quickly and accurately evaluated, which provides a scientific basis for early intervention, clinical diagnosis or personalized medicine of individual aging and / or cognitive impairment, and has a wide application prospect.

[0050] In the present application, the term "CYP2C family gene" is derived from the human genome, and refers to a specific group of genes located on chromosome 10q24 in the human cytochrome P450 superfamily, which encode enzymes involved in drug metabolism, hormone synthesis and decomposition, and cholesterol metabolism and other biochemical processes. The CYP2C family gene includes multiple subtypes, such as CYP2C8, CYP2C9, CYP2C18 and CYP2C19, etc.

[0051] In the present application, the term "Cyp2c family gene" is derived from the genome of non-human species, such as mice, rats, etc., and refers to cytochrome P450 genes in non-human species that are homologous or have similar functions to human CYP2C family genes. These genes also encode cytochrome P450 enzymes and are involved in the physiological and biochemical processes of their respective species. For example, in mice, the Cyp2c family genes include Cyp2c27, Cyp2c37, Cyp2c40, etc.

[0052] In the present application, the term "expression product" refers to a protein or functional molecule produced by a specific gene through the transcription and translation process, including proteins, etc.

[0053] According to an embodiment of the present application, the CYP2C family gene comprises one or more of CYP2C8, CYP2C9, CYP2C18 and CYP2C19. Thus, when the target detection object is human, multiple markers are provided for selection, and researchers can select the specific CYP2C family gene with the best detection effect as the marker according to different research needs and clinical application scenarios.

[0054] According to an embodiment of the present application, the Cyp2c family genes include one or more of Cyp2c29, Cyp2c38, Cyp2c37, Cyp2c40, Cyp2c50 and Cyp2c54. Thus, when the target detection object is an experimental animal such as a mouse, and a variety of markers are provided for selection, researchers can select a specific Cyp2c family gene with the best detection effect as a marker according to different research needs, specific detection objects and clinical application scenarios.

[0055] According to a preferred embodiment of the present application, the CYP2C family gene is CYP2C9 and CYP2C19. Thus, the sensitivity and specificity of the marker are further optimized.

[0056] According to a preferred embodiment of the present application, the Cyp2c family gene is Cyp2c29. Thus, the sensitivity and specificity of the marker are further optimized.

[0057] It should be noted that although the embodiments of the present application only use mice as experimental animals and use Cyp2c29 genes and their expression products from the experimental animal as markers to verify their functions in evaluating the degree of aging and the risk of cognitive impairment of individuals, other genes in the Cyp2c family such as Cyp2c38, Cyp2c37, Cyp2c40, Cyp2c50 and Cyp2c54 have similar biological functions, regulatory mechanisms and potential roles in aging and cognitive impairment as Cyp2c29 genes. In addition, human CYP2C family genes homologous to Cyp2c family genes, such as CYP2C8, CYP2C9, CYP2C18 and CYP2C19, also have similar biological functions, regulatory mechanisms and potential roles in aging and cognitive impairment. Therefore, CYP2C family genes including Cyp2c29 genes and their expression products, Cyp2c family genes and their expression products should all fall within the protection scope of the present application.

[0058] It should be noted that the above only lists several common CYP2C family genes / Cyp2c family genes, but as the study of genomics continues to deepen, more CYP2C family genes and Cyp2c family genes related to aging and cognitive impairment may be discovered. Therefore, CYP2C family genes and their expression products and Cyp2c family genes and their expression products, whether or not specifically mentioned in the present application, should all fall within the protection scope of the present application.

[0059] Use in the preparation of a kit

[0060] The present application provides CYP2C family genes and expression products thereof and / or Cyp2c family genes and expression products thereof, and the use of reagents for detecting CYP2C family genes and expression products thereof and / or Cyp2c family genes and expression products thereof in the preparation of a kit. According to embodiments of the present application, the kit is used for detecting aging and / or cognitive impairment.

[0061] It will be appreciated by those skilled in the art that the features and advantages described above for the marker for detecting aging and / or cognitive impairment also apply to the use, which will not be described again here.

[0062] According to embodiments of the present application, a decrease in the level of CYP2C family genes and expression products thereof and / or Cyp2c family genes and expression products thereof is indicative of aging and / or cognitive impairment. Thus, by detecting the level of this highly specific and sensitive marker, CYP2C family genes and expression products thereof and / or Cyp2c family genes and expression products thereof, the degree of aging and the risk of cognitive impairment of an individual can be quickly and accurately assessed.

[0063] According to embodiments of the present application, the CYP2C family genes include one or more of CYP2C8, CYP2C9, CYP2C18 and CYP2C19.

[0064] According to embodiments of the present application, the Cyp2c family genes include one or more of Cyp2c29, Cyp2c38, Cyp2c37, Cyp2c40, Cyp2c50 and Cyp2c54.

[0065] According to preferred embodiments of the present application, the CYP2C family genes are CYP2C9 and CYP2C19.

[0066] According to preferred embodiments of the present application, the Cyp2c family genes are Cyp2c29.

[0067] It should be noted that the above only lists several common CYP2C family genes / Cyp2c family genes, but as the study of genomics continues to deepen, more CYP2C family genes and Cyp2c family genes related to aging and cognitive impairment may be discovered, and therefore, CYP2C family genes and expression products thereof and Cyp2c family genes and expression products thereof, whether or not specifically mentioned in the present application, shall fall within the scope of protection of the present application.

[0068] Kit for detecting aging and / or cognitive impairment

[0069] The present application provides a kit for detecting aging and / or cognitive impairment. According to an embodiment of the present application, the kit contains reagents for detecting CYP2C family genes and their expression products and / or Cyp2c family genes and their expression products. According to an embodiment of the present application, the kit can detect the expression levels of CYP2C family genes and their expression products and / or Cyp2c family genes and their expression products. Further, the use of the kit of the present application for detecting the sample to be tested can quickly and accurately assess the degree of aging and the risk of cognitive impairment of an individual, and has a wide application prospect.

[0070] According to an embodiment of the present application, the CYP2C family genes include one or more of CYP2C8, CYP2C9, CYP2C18 and CYP2C19. Thus, the target detection object of the kit is human at this time, and multiple markers are provided for selection, and researchers can select the specific CYP2C family genes with the best detection effect as markers according to different research needs and clinical application scenarios.

[0071] According to an embodiment of the present application, the Cyp2c family genes include one or more of Cyp2c29, Cyp2c38, Cyp2c37, Cyp2c40, Cyp2c50 and Cyp2c54. Thus, the target detection object of the reagent or kit is an experimental animal such as a mouse at this time, and multiple markers are provided for selection, and researchers can select the Cyp2c family genes with the best detection effect as markers according to different research needs, specific detection objects and clinical application scenarios.

[0072] According to a preferred embodiment of the present application, the CYP2C family genes are CYP2C9 and CYP2C19. Thus, the sensitivity and specificity of the markers are further optimized, thereby improving the detection specificity and sensitivity of the kit.

[0073] According to a preferred embodiment of the present application, the Cyp2c family genes are Cyp2c29. Thus, the sensitivity and specificity of the markers are further optimized, thereby improving the detection specificity and sensitivity of the kit.

[0074] It should be noted that the "sample to be tested" of the present application can be from a healthy individual or a patient or a subject or an experimental animal, including but not limited to blood, urine, saliva, feces, tissue samples, cell samples, cerebrospinal fluid, semen, amniotic fluid or other body fluids such as joint cavity fluid, pleural effusion, etc.; or other biological materials, including but not limited to cells obtained by biological technology means.

[0075] Method for constructing an animal model

[0076] The application provides a method for constructing an animal model. According to an embodiment of the application, the method comprises the following steps: editing a genome of a target animal by using genetic engineering technology to modify a Cyp2c family gene, and obtaining a target animal model. According to the method, researchers can construct an animal model in which a Cyp2c family gene is silenced, so as to further study the mechanism of the Cyp2c family gene in aging and cognitive impairment, and further explore the potential functions and application values of a homologous gene (CYP2C family gene) of the Cyp2c family gene in human aging and cognitive impairment, and provide an experimental basis for early intervention, personalized medicine or new treatment strategy development for human individual aging and / or cognitive impairment.

[0077] According to an embodiment of the application, the Cyp2c family gene comprises one or more of Cyp2c29, Cyp2c38, Cyp2c37, Cyp2c40, Cyp2c50 and Cyp2c54. In this way, researchers can select Cyp2c family genes for experiments according to different research needs, specific detection objects and clinical application scenarios; for example, researchers can select different Cyp2c family genes for modification (including knockdown, knockout, overexpression, etc.) to further explore the specific mechanisms of the Cyp2c family gene and its homologous gene CYP2C family gene in aging and cognitive impairment, and further provide a theoretical basis for the development of new treatment strategies for cognitive impairment and other diseases.

[0078] According to an embodiment of the application, the modification comprises one or more of knockdown, knockout, overexpression, gene silencing and gene activation. In this way, researchers can select appropriate gene modification methods according to specific experimental purposes to achieve fine regulation of the function of the Cyp2c family gene; for example, knockout or gene silencing can be used to study the effects of Cyp2c family gene deletion on aging and cognitive impairment, while overexpression or gene activation can be used to explore the potential effects of enhanced Cyp2c family gene function on the aging process. By using different modification methods, researchers can further study the mechanism of the Cyp2c family gene in aging and cognitive impairment from multiple angles.

[0079] It should be noted that the above only lists several common gene modification methods, but the protection scope of the present application is not limited thereto, with the continuous development of genetic engineering technology, new gene modification methods and technical means are constantly emerging, for example, gene editing technology (such as CRISPR / Cas9), RNA interference technology (RNAi), antisense oligonucleotide technology (ASO), gene knock-in (Gene Knock-in) and the like, these methods can also be used to modify Cyp2c family genes to achieve the research on the mechanism related to aging and cognitive impairment, therefore, all gene modification methods that can realize the functional regulation of Cyp2c family genes, whether or not specifically mentioned in the present application, shall belong to the protection scope of the present application.

[0080] According to an embodiment of the present application, the target animals include one or more of mice, rats, rabbits, pigs and non-human primates. Thus, the diversified selection of target animals provides researchers with a wide range of experimental platforms, which can meet the needs of different research stages; for example, mice and rats have the advantages of fast reproduction, low cost, clear genetic background, etc., and can be applied to preliminary Cyp2c family gene function research and drug screening.

[0081] Application in drug development or preparation

[0082] The present application proposes the application of CYP2C family genes and expression products thereof and / or Cyp2c family genes and expression products thereof, the marker of the first aspect and the kit of the third aspect in the development or preparation of drugs for preventing, alleviating, assisting treatment or treating cognitive dysfunction.

[0083] It can be understood by those skilled in the art that the features and advantages described above for the marker or kit also apply to the application, which will not be repeated here.

[0084] According to an embodiment of the present application, the CYP2C family genes include one or more of CYP2C8, CYP2C9, CYP2C18 and CYP2C19.

[0085] According to an embodiment of the present application, the Cyp2c family genes include one or more of Cyp2c29, Cyp2c38, Cyp2c37, Cyp2c40, Cyp2c50 and Cyp2c54.

[0086] It should be noted that the above only lists several common CYP2C family genes / Cyp2c family genes, but as the study of genomics continues to deepen, more CYP2C family genes and Cyp2c family genes related to aging and cognitive impairment may be discovered, therefore, the CYP2C family genes and their expression products and the Cyp2c family genes and their expression products, whether or not specifically mentioned in this application, shall belong to the protection scope of the present application.

[0087] The scheme of the present application will be explained below in combination with examples. Those skilled in the art will understand that the following examples are only for illustrating the present application and should not be regarded as limiting the scope of the present application. If a specific technique or condition is not specified in the examples, it is carried out according to the technique or condition described in the literature in the art or according to the product instruction. If the reagent or instrument used is not specified by the manufacturer, it is a conventional product that can be obtained by purchase in the market.

[0088] Example 1: Significant down-regulation of CYP-related metabolites / genes in the liver of naturally aged mice

[0089] 1. Construction of mouse model

[0090] (1) Control group (Young): 8 C57BL / 6 male mice were normally fed with ordinary feed until 2 months old to obtain control group mice A1.

[0091] (2) Experimental group (Old): 8 C57BL / 6 male mice were normally fed with ordinary feed until 18 months old to obtain experimental group mice A2 (naturally aged model mice).

[0092] 2. Sample collection

[0093] After euthanasia of the control group mice A1 and the experimental group mice A2, the liver tissues of the mice were collected, a part of the liver tissues was used to make pathological sections and fixed in 4% paraformaldehyde solution, and the other part was quickly frozen in liquid nitrogen and stored at -80℃ for standby.

[0094] 3. Lipid metabolism sequencing analysis

[0095] The lipid samples of the liver tissues of the control group mice A1 and the experimental group mice A2 were extracted respectively for lipid metabolism sequencing analysis.

[0096] 4. Gene expression analysis

[0097] According to the results of lipid metabolism sequencing analysis, the expression levels of Cyp2c family related genes in the control group mice A1 and the experimental group mice A2 liver tissue samples collected in step 3 were detected by RT-qPCR technology.

[0098] The results of lipid metabolism sequencing analysis and gene expression analysis of naturally aged mice are shown in Figure 1 .

[0099] The results show that:

[0100] (1) The results of lipid metabolism sequencing analysis show that, compared with the control group mice A1, the levels of metabolites related to the cyclooxygenase (COX), lipoxygenase (LOX) and cytochrome P450 (CYP) pathways in the liver of the experimental group mice A2 are significantly down-regulated, among which the CYP pathway has the most down-regulated metabolites, accounting for 51.73% of the total down-regulated metabolites (Fig. 1B and C). Figure 1

[0101] (2) The results of gene expression analysis show that, compared with the control group mice A1, the mRNA expression levels of Cyp2c29, Cyp2c38, Cyp2c40, Cyp2c50 and Cyp2c54 genes in the liver of the experimental group mice A2 are significantly reduced (Fig. 1D). Figure 1

[0102] Example 2: Significant down-regulation of CYP-related genes in the liver of chemically induced aging mice

[0103] 1. Construction of mouse model

[0104] (1) Control group (Control): 12 8-week-old C57BL / 6 male mice were subcutaneously injected with 0.9% normal saline every day for 10 consecutive weeks (0.2 mL of 0.9% normal saline was subcutaneously injected per 20 g of body weight according to the body weight of the mice), obtaining control group mice B1.

[0105] (2) Experimental group (D-gal): 12 8-week-old C57BL / 6 male mice were subcutaneously injected with 200 mg / kg D-galactose (D-gal) every day for 10 consecutive weeks, obtaining experimental group mice B2.

[0106] 2. Sample collection

[0107] After euthanizing the control group mice B1 and the experimental group mice B2, the liver tissues of the mice were collected, a part of the liver tissues were used to make pathological sections and fixed in 4% paraformaldehyde solution, and the other part was quickly frozen in liquid nitrogen and stored at -80°C for standby.

[0108] 3. Gene expression analysis

[0109] The expression levels of Cyp2c family-related genes in the liver tissue samples of the control group mice B1 and the experimental group mice B2 collected in step 2 were detected by RT-qPCR technology.

[0110] ​​The results of sequencing analysis and gene expression analysis of lipid metabolism in chemically-induced aging mice are shown in Figure 2 .

[0111] The results show that, compared with the control group mice B1, the mRNA expression levels of Cyp2c29, Cyp2c38, Cyp2c40, Cyp2c50 and Cyp2c54 genes in the liver of the experimental group mice B2 induced by D-gal are significantly reduced.

[0112] Example 3: Cyp2c29 gene deletion accelerates cognitive impairment

[0113] Through sequence alignment analysis, it is found that the mouse Cyp2c29 gene has very high similarity with human genes CYP2C9 and CYP2C19, wherein the isozyme sequence similarity of the mouse Cyp2c29 gene with the human gene CYP2C9 is 76.01% (Table 1), and the sequence similarity of the mouse Cyp2c29 gene with the human gene CYP2C19 is 75.00% (Table 2). Further, the mouse Cyp2c29 gene can be used as an important model for studying the functions of human genes CYP2C9 and CYP2C19. Based on this, in order to explore the effects of Cyp2c related genes on the cognitive function of the body in aging, the inventors constructed a Cyp2c29 gene knockout (KO) mouse model using C57BL / 6N wild-type mice, and carried out related exploratory experiments, the specific steps are as follows:

[0114] 1. Construction of Cyp2c29 gene knockout mouse model

[0115] (1) Control group (WT): 10 28-week-old C57BL / 6N wild-type male mice were subcutaneously injected with 200 mg / kg D-galactose every day for 10 consecutive weeks to obtain control group mice C1.

[0116] (2) Experimental group (Cyp2c29 KO): 10 28-week-old Cyp2c29 KO male mice were subcutaneously injected with 200 mg / kg D-galactose every day for 10 consecutive weeks to obtain experimental group mice C2.

[0117] 2. Grip strength and behavioral detection

[0118] In step 1, the Cyp2c29 gene knockout mouse model construction was carried out to the 9th week, and the control group mice C1 and the experimental group mice C2 were used for grip strength and behavioral detection, wherein the behavioral detection included three parts:

[0119] (1) Open-field test

[0120] Record the total distance and rapid movement speed of control group mice C1 and experimental group mice C2 in the open field.

[0121] (2) Novel object recognition test

[0122] Place control group mice C1 and experimental group mice C2 in an environment containing two identical objects, and record the exploration time of the two groups of mice to the objects; 1 h later, replace one of the objects with a new object, and record the exploration time of the two groups of mice to the new object and the old object.

[0123] (3) Y-maze test

[0124] Record the autonomous alternation ratio and novel arm residence time of control group mice C1 and experimental group mice C2 in the Y-maze.

[0125] The sequence alignment results of human gene CYP2C9 and mouse Cyp2c family gene isozyme are shown in Table 1, the sequence alignment results of human gene CYP2C19 and mouse Cyp2c family gene isozyme are shown in Table 2, and the grip strength ability and behavioral detection results of WT mice and Cyp2c29 KO mice are shown in Figure 3 .

[0126] Table 1 Sequence alignment results of human gene CYP2C9 and mouse Cyp2c family gene isozyme

[0127]

[0128]

[0129] Table 2 Sequence alignment results of human gene CYP2C19 and mouse Cyp2c family gene isozyme

[0130] Mice isoenzyme Max score E value Identification (%) Cyp2c29 771 0.0 75.00 Cyp2c37 726 0.0 72.60 Cyp2c38 741 0.0 72.46 Cyp2c39 751 0.0 73.89 Cyp2c40 659 0.0 66.33 Cyp2c44 630 0.0 63.69 Cyp2c50 725 0.0 72.60 Cyp2c54 704 0.0 70.76 Cyp2c55 730 0.0 72.80 Cyp2c65 655 0.0 72.33 Cyp2c67 661 0.0 66.73 Cyp2c68 662 0.0 66.53 Cyp2c69 664 0.0 66.73 Cyp2c70 634 0.0 62.08

[0131] In Table 1 and Table 2, Mice isozyme refers to mouse isozyme; Max score refers to the highest alignment score in the BLAST sequence alignment result; E value refers to an important indicator in the BLAST sequence alignment result, which represents the number of times that the same or better alignment is expected to be obtained under random conditions. The lower the E value, the more significant the alignment result, i.e. the less likely it is to be obtained by chance. An alignment with an E value close to zero is considered significant; Identification (%) refers to the percentage of homology between mouse sequence and human sequence, i.e. the proportion of identical nucleotides or amino acids when the query sequence is aligned with the database sequence.

[0132] The results show that: compared with the D-gal induced wild type (WT) aging mice (control group mice C1), the total distance and the fast movement speed of D-gal induced Cyp2c29 KO aging mice (experimental group mice C2) significantly decreased; at the same time, the ability of the mice to recognize new objects also significantly decreased; in addition, in the Y maze experiment, the self-alternation ratio and the new strange arm residence time of the mice also significantly decreased.

[0133] The above results show that: Cyp2c29 gene is closely related to the cognitive level of mice, and the deletion of Cyp2c29 gene will significantly reduce the spatial learning and memory ability of mice, leading to the occurrence of cognitive impairment, and it can be inferred that human CYP2C family genes (CYP2C9 and CYP2C19, etc.) are closely related to the cognitive level of human body.

[0134] In the description of the present specification, the description of the terms "one embodiment", "some embodiments", "an example", "a specific example", or "some examples" and the like means that the specific features, structures, materials or characteristics described in connection with the embodiment or example are included in at least one embodiment or example of the present application. In the present specification, the illustrative description of the above terms does not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any appropriate manner in any one or more embodiments or examples. In addition, different embodiments or examples described in the present specification and the features of different embodiments or examples can be combined and combined by those skilled in the art without contradiction.

[0135] Although the embodiments of the present application have been shown and described above, it should be understood that the above embodiments are exemplary and should not be construed as limiting the present application, and those skilled in the art can make changes, modifications, replacements and variations to the above embodiments within the scope of the present application.

Claims

1. A biomarker for detecting aging and / or cognitive impairment, characterized in that... The biomarkers include CYP2C family genes and their expression products and / or Cyp2c family genes and their expression products.

2. The marker according to claim 1, characterized in that, The CYP2C family genes include one or more of CYP2C8, CYP2C9, CYP2C18, and CYP2C19; Optionally, the Cyp2c family genes include one or more of Cyp2c29, Cyp2c38, Cyp2c37, Cyp2c40, Cyp2c50 and Cyp2c54; Preferably, the CYP2C family genes are CYP2C9 and CYP2C19; Preferably, the Cyp2c family gene is Cyp2c29.

3. The use of CYP2C family genes and their expression products and / or Cyp2c family genes and their expression products, and reagents for detecting CYP2C family genes and their expression products and / or Cyp2c family genes and their expression products in the preparation of a kit, characterized in that, The kit is used to detect aging and / or cognitive impairment.

4. The use according to claim 3, characterized in that, Decreased levels of the CYP2C family genes and their expression products and / or Cyp2c family genes and their expression products are indicators of aging and / or cognitive impairment. Optionally, the CYP2C family genes include one or more of CYP2C8, CYP2C9, CYP2C18, and CYP2C19; Optionally, the Cyp2c family genes include one or more of Cyp2c29, Cyp2c38, Cyp2c37, Cyp2c40, Cyp2c50 and Cyp2c54; Preferably, the CYP2C family genes are CYP2C9 and CYP2C19; Preferably, the Cyp2c family gene is Cyp2c29.

5. A kit for detecting aging and / or cognitive impairment, characterized in that, The kit contains reagents for detecting CYP2C family genes and their expression products and / or Cyp2c family genes and their expression products.

6. The reagent kit according to claim 5, characterized in that, The CYP2C family genes include one or more of CYP2C8, CYP2C9, CYP2C18, and CYP2C19; Optionally, the Cyp2c family genes include one or more of Cyp2c29, Cyp2c38, Cyp2c37, Cyp2c40, Cyp2c50 and Cyp2c54; Preferably, the CYP2C family genes are CYP2C9 and CYP2C19; Preferably, the Cyp2c family gene is Cyp2c29.

7. A method for constructing an animal model, characterized in that, Includes the following steps: Genetic engineering techniques were used to edit the genome of the target animal to modify Cyp2c family genes, thereby obtaining the target animal model.

8. The construction method according to claim 7, characterized in that, The Cyp2c family of genes includes one or more of Cyp2c29, Cyp2c38, Cyp2c37, Cyp2c40, Cyp2c50, and Cyp2c54; Optionally, the modification includes one or more of knockdown, knockout, overexpression, gene silencing, and gene activation.

9. The construction method according to claim 7, characterized in that, The target animals include one or more of the following: mice, rats, rabbits, pigs, and non-human primates.

10. The use of CYP2C family genes and their expression products and / or Cyp2c family genes and their expression products, the biomarkers of claim 1 or 2, and the kits of claim 5 or 6 in the development or preparation of medicaments for the prevention, relief, adjunctive treatment or treatment of cognitive impairment.

11. The application according to claim 10, characterized in that, The CYP2C family genes include one or more of CYP2C8, CYP2C9, CYP2C18, and CYP2C19; Optionally, the Cyp2c family genes include one or more of Cyp2c29, Cyp2c38, Cyp2c37, Cyp2c40, Cyp2c50 and Cyp2c54.

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