CN113880934A - Skin rejuvenation protein marker LRC59 protein and noninvasive extraction method thereof - Google Patents
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Abstract
The invention discloses a skin rejuvenation protein marker LRC59 protein and a noninvasive extraction method thereof. The application of LRC59 protein in auxiliary judgment of skin rejuvenation degree and an extraction and detection method are disclosed, aiming at finding out the intrinsic factor of skin rejuvenation from the root, intervening skin aging in advance before the appearance of skin aging, keeping the skin rejuvenated, and in addition, correctly judging the skin rejuvenation degree of the skin and judging whether the physiological age of skin rejuvenation is consistent with the actual age, thereby providing reference and direction for beauty treatment or medical beauty treatment.
Description
Technical Field
The invention relates to the field of molecular biology, in particular to a skin rejuvenation protein marker LRC59 protein and a noninvasive extraction method thereof.
Background
Criteria for skin rejuvenation: the sebum secretion is moderate, the skin is neither dry nor oily, the skin color is ruddy and fine, the skin is elastic, the thickness is moderate, pores are small, the skin is insensitive to external stimulation, and the pH value is 5-5.6.
With the improvement of living standard, people pay more and more attention to skin care, but usually only pay attention to the external manifestations of skin aging, such as wrinkle, color spot, pore thickness degree and other information, and judge the youth degree of skin from the information.
LRC59 (leucine rich repeat protein 59) is a tail ankyrin with a single transmembrane domain near its C-terminus that localizes to the Endoplasmic Reticulum (ER) and the nuclear envelope. LRRC59 may reduce TLR3, 8 and 9 mediated signaling, but not TLR4 mediated signaling. After stimulation of ligand, LRRC59 was TLR independent of its association with UNC93B1, which required ligand internalization induced signaling. Endosomal localization of endogenous TLR3 was reduced by silencing LRRC59, suggesting that LRRC59 promotes ER-mediated translocation of UNC93B 1-mediated NA-responsive TLRs from the ER following infection. Are important positive modulators of DDX 58-mediated type I IFN signaling. LRRC59 interacts specifically with ISG 15-associated DDX58 and prevents its binding to LRRC25 after viral infection, LRRC25 is a secondary receptor that delivers DDX58 to SQSTM1/p 62-dependent degradation of autophagosomes, leading to a stronger antiviral immune response, and DDX58/RIG-I is a key pattern recognition receptor for viral RNA, plays a crucial role in antiviral immunity, and maintains immune homeostasis.
There is currently no precedent for using LRC59 protein (leucine rich repeat inclusion 59) to aid in determining skin youth.
Disclosure of Invention
A method for non-invasive extraction of LRC59 protein from skin, the method comprising the steps of:
(1) sampling of skin samples of the epidermis of a subject: sticking the 3M medical adhesive patch to the curved side part of the forearm, and slightly removing the 3M adhesive patch after 1 minute to obtain a sticky tape-shaped skin sample;
(2) obtaining of a dried peptide fragment sample: 1) cutting the adhesive tape-shaped skin sample into small pieces, depositing on a glass plate, and transferring to a centrifuge tube;
2) adding a proper amount of lysis buffer sample without SDS, adding 2mM EDTA and 1XCocktail, then placing on ice for 5 minutes, then adding 10mM DTT, and soaking the sample overnight;
3) centrifuging at 25,000g centrifugal force at 4 deg.C for 15 minutes, recovering the supernatant and treating DTT with 10mM for 1 hour in a water bath at 56 deg.C;
4) then treated with 55mM IAM, incubated for 45 minutes at room temperature in the dark, and centrifuged at 25,000g at 4 ℃ for 15 minutes to give the final protein solution supernatant; protein concentration was measured using the Bradford method, and extracted proteins were quality-controlled by 12% SDS-PAGE; taking 100 μ g of protein from each sample, adding trypsin and hydrolyzing at 37 deg.C for 4 hr; then adding trypsin again in the same proportion for enzymolysis for 8 hours at 37 ℃; desalting the polypeptide with Strata X chromatographic column and vacuum drying to obtain dried peptide sample.
Preferably, the method for determining the relative content of LRC59 protein in the epidermal skin sample based on mass spectrum comprises the following steps: (1) sampling of skin samples of the epidermis of a subject: sticking the 3M medical adhesive patch to the curved side part of the forearm, and slightly removing the 3M adhesive patch after 1 minute to obtain a sticky tape-shaped skin sample;
(2) obtaining of a dried peptide fragment sample: 1) cutting the adhesive tape-shaped skin sample into small pieces, depositing on a glass plate, and transferring to a centrifuge tube;
2) adding a proper amount of lysis buffer sample without SDS, adding 2mM EDTA and 1XCocktail, then placing on ice for 5 minutes, then adding 10mM DTT, and soaking the sample overnight;
3) centrifuging at 25,000g centrifugal force at 4 deg.C for 15 minutes, recovering the supernatant and treating DTT with 10mM for 1 hour in a water bath at 56 deg.C;
4) then treated with 55mM IAM, incubated for 45 minutes at room temperature in the dark, and centrifuged at 25,000g at 4 ℃ for 15 minutes to give the final protein solution supernatant; protein concentration was measured using the Bradford method, and extracted proteins were quality-controlled by 12% SDS-PAGE; taking 100 μ g of protein from each sample, adding trypsin and hydrolyzing at 37 deg.C for 4 hr; then adding trypsin again in the same proportion for enzymolysis for 8 hours at 37 ℃; desalting the polypeptide with Strata X chromatographic column and vacuum drying to obtain dried peptide sample;
(3) detection of
Redissolving the dried peptide fragment sample with mobile phase A (2% ACN, 0.1% FA), centrifuging at 20,000g for 10 min, and sampling the supernatant; separation by ULRC59 protein LC; the sample was first enriched and desalted on a trap column, then connected in series with a self-contained C18 column, at a flow rate of 500nl/min, by the following effective gradient:
separation: 0-5min, 5% mobile phase B (98% ACN, 0.1% FA); 5-160min, mobile phase B increased linearly from 5% to 35%; 160-170min, the mobile phase B rises from 35% to 80%; 170 ℃ 175min, 80% mobile phase B; 176 ℃ for 180min, 5% of mobile phase B; the end of the nanoliter liquid phase separation is directly connected with a mass spectrometer;
DDA mass spectrometric detection
The peptide segment separated by the liquid phase is ionized by a nanoESI source and then is imported to a tandem mass spectrometer Q-active HF mode for detection; setting main parameters: the ion source voltage was set to 1.6 kV; the primary mass spectrum scanning range is 350-1500 m/z; resolution was set to 60,000; the initial m/z of the secondary mass spectrum is fixed to be 100; resolution 15,000. The screening conditions of the parent ions for secondary fragmentation are as follows: parent ions with charges 2+ to 7+, with intensities in excess of 10,000 peak intensity ranked first 20; the ion fragmentation mode is HCD, and fragment ions are detected in Orbitrap; the dynamic exclusion time was set to 30 s; the AGC is set as: primary 3E6, secondary 1E 5;
DIA mass spectrometric detection
The peptide segment separated by the liquid phase is ionized by a nanoESI source and then is imported to a tandem mass spectrometer Q-active HF mode for detection; setting main parameters: the ion source voltage was set to 1.6 kV; the primary mass spectrum scanning range is 350-1500 m/z; resolution was set to 120,000; uniformly dividing 350-1500Da into 40 windows for fragmentation and signal acquisition; the ion fragmentation mode is HCD, and fragment ions are detected in Orbitrap; the dynamic exclusion time was set to 30 s; the AGC is set as: primary 3E6, secondary 1E 5.
Preferably, the substance for detecting the content of LRC59 protein is a mass spectrometric identification reagent, an antibody or an antigen-binding fragment thereof; the substance for detecting the content of the LRC59 protein is an orbital trap high-resolution mass spectrometer.
Preferably, the P value of the LRC59 protein is 0.034750959.
Preferably, the method for judging the aging degree and the skin aging degree by the LRC59 protein comprises the following steps:
1) taking a sample of the epidermal skin of a subject;
2) detecting the content of LRC59 protein in the skin sample of the obtained subject;
3) comparing the LRC59 protein content measured in the step 2) with the LRC59 protein content value in the skin of the person with normal aging at the age group, and judging the skin aging degree of the subject according to the comparison result;
or 4) comparing the LRC59 protein content measured in the step 2) with the LRC59 protein content standard curve in the skin of the normally aged person of each age group, and judging the physiological age of the skin of the subject according to the comparison result.
Preferably, the system for assisting in determining the degree of aging comprises the following modules:
(1) a data receiving module; the data receiving module is configured to receive LRC59 protein content data in a skin sample of a subject;
(2) a data storage module: the data storage module is configured to store LRC59 protein content data in normal human skin consistent with the age bracket of the subject;
(3) a data comparison module: the data comparison module is configured to compare the LRC59 protein content data in the skin sample of the subject received by the data receiving module with LRC59 protein content data in normal human skin which is consistent with the age group of the subject and stored in the data storage module;
(4) a judgment module; the judging module is configured to receive the comparison result sent by the data comparing module, judge the comparison result, judge the skin aging degree of the subject, or judge whether the skin physiological age of the subject is consistent with the actual age of the subject, and output the judgment result.
The method for detecting the LRC59 protein in the skin to assist in judging the skin youth degree is simple, accurate in result and high in efficiency. The method is characterized in that the intrinsic factors causing skin aging are found out radically, the skin aging is intervened in advance before the appearance of the external appearance of the skin aging, in addition, the youth degree of the skin can be judged correctly, whether the physiological age of the aging is consistent with the actual age can be judged, and a reference and a direction are provided for beauty treatment or medical beauty treatment.
Drawings
FIG. 1 is a mass spectrum of a characteristic peptide fragment (LQQLPADFGR) of LRC59 protein obtained by detection.
Detailed Description
The present invention is further described below by way of specific examples, but the present invention is not limited to only the following examples. Variations, combinations, or substitutions of the invention, which are within the scope of the invention or the spirit, scope of the invention, will be apparent to those of skill in the art and are within the scope of the invention.
The experimental methods used in the following examples are all conventional methods unless otherwise specified; reagents, materials and the like used in the following examples are commercially available unless otherwise specified.
A method for non-invasive extraction of LRC59 protein from skin, the method comprising the steps of:
(1) sampling of skin samples of the epidermis of a subject: sticking the 3M medical adhesive patch to the curved side part of the forearm, and slightly removing the 3M adhesive patch after 1 minute to obtain a sticky tape-shaped skin sample;
(2) obtaining of a dried peptide fragment sample: 1) cutting the adhesive tape-shaped skin sample into small pieces, depositing on a glass plate, and transferring to a centrifuge tube;
2) adding a proper amount of lysis buffer sample without SDS, adding 2mM EDTA and 1XCocktail, then placing on ice for 5 minutes, then adding 10mM DTT, and soaking the sample overnight;
3) centrifuging at 25,000g centrifugal force at 4 deg.C for 15 minutes, recovering the supernatant and treating DTT with 10mM for 1 hour in a water bath at 56 deg.C;
4) then treated with 55mM IAM, incubated for 45 minutes at room temperature in the dark, and centrifuged at 25,000g at 4 ℃ for 15 minutes to give the final protein solution supernatant; protein concentration was measured using the Bradford method, and extracted proteins were quality-controlled by 12% SDS-PAGE; taking 100 μ g of protein from each sample, adding trypsin and hydrolyzing at 37 deg.C for 4 hr; then adding trypsin again in the same proportion for enzymolysis for 8 hours at 37 ℃; desalting the polypeptide with Strata X chromatographic column and vacuum drying to obtain dried peptide sample.
The method for measuring the relative content of LRC59 protein in an epidermal skin sample based on mass spectrum comprises the following steps: (1) sampling of skin samples of the epidermis of a subject: sticking the 3M medical adhesive patch to the curved side part of the forearm, and slightly removing the 3M adhesive patch after 1 minute to obtain a sticky tape-shaped skin sample;
(2) obtaining of a dried peptide fragment sample: 1) cutting the adhesive tape-shaped skin sample into small pieces, depositing on a glass plate, and transferring to a centrifuge tube;
2) adding a proper amount of lysis buffer sample without SDS, adding 2mM EDTA and 1XCocktail, then placing on ice for 5 minutes, then adding 10mM DTT, and soaking the sample overnight;
3) centrifuging at 25,000g centrifugal force at 4 deg.C for 15 minutes, recovering the supernatant and treating DTT with 10mM for 1 hour in a water bath at 56 deg.C;
4) then treated with 55mM IAM, incubated for 45 minutes at room temperature in the dark, and centrifuged at 25,000g at 4 ℃ for 15 minutes to give the final protein solution supernatant; protein concentration was measured using the Bradford method, and extracted proteins were quality-controlled by 12% SDS-PAGE; taking 100 μ g of protein from each sample, adding trypsin and hydrolyzing at 37 deg.C for 4 hr; then adding trypsin again in the same proportion for enzymolysis for 8 hours at 37 ℃; desalting the polypeptide with Strata X chromatographic column and vacuum drying to obtain dried peptide sample;
(3) detection of
Redissolving the dried peptide fragment sample with mobile phase A (2% ACN, 0.1% FA), centrifuging at 20,000g for 10 min, and sampling the supernatant; separation by ULRC59 protein LC; the sample was first enriched and desalted on a trap column, then connected in series with a self-contained C18 column, at a flow rate of 500nl/min, by the following effective gradient:
separation: 0-5min, 5% mobile phase B (98% ACN, 0.1% FA); 5-160min, mobile phase B increased linearly from 5% to 35%; 160-170min, the mobile phase B rises from 35% to 80%; 170 ℃ 175min, 80% mobile phase B; 176 ℃ for 180min, 5% of mobile phase B; the end of the nanoliter liquid phase separation is directly connected with a mass spectrometer;
DDA mass spectrometric detection
The peptide segment separated by the liquid phase is ionized by a nanoESI source and then is imported to a tandem mass spectrometer Q-active HF mode for detection; setting main parameters: the ion source voltage was set to 1.6 kV; the primary mass spectrum scanning range is 350-1500 m/z; resolution was set to 60,000; the initial m/z of the secondary mass spectrum is fixed to be 100; resolution 15,000. The screening conditions of the parent ions for secondary fragmentation are as follows: parent ions with charges 2+ to 7+, with intensities in excess of 10,000 peak intensity ranked first 20; the ion fragmentation mode is HCD, and fragment ions are detected in Orbitrap; the dynamic exclusion time was set to 30 s; the AGC is set as: primary 3E6, secondary 1E 5;
DIA mass spectrometric detection
The peptide segment separated by the liquid phase is ionized by a nanoESI source and then is imported to a tandem mass spectrometer Q-active HF mode for detection; setting main parameters: the ion source voltage was set to 1.6 kV; the primary mass spectrum scanning range is 350-1500 m/z; resolution was set to 120,000; uniformly dividing 350-1500Da into 40 windows for fragmentation and signal acquisition; the ion fragmentation mode is HCD, and fragment ions are detected in Orbitrap; the dynamic exclusion time was set to 30 s; the AGC is set as: primary 3E6, secondary 1E 5.
The substance for detecting the content of the LRC59 protein is a mass spectrometric identification reagent, an antibody or an antigen-binding fragment thereof; the substance for detecting the content of the LRC59 protein is an orbital trap high-resolution mass spectrometer.
The P value of the LRC59 protein is 0.034750959.
The method for judging the aging degree and the skin aging degree by the LRC59 protein comprises the following steps:
1) taking a sample of the epidermal skin of a subject;
2) detecting the content of LRC59 protein in the skin sample of the obtained subject;
3) comparing the LRC59 protein content measured in the step 2) with the LRC59 protein content value in the skin of the person with normal aging at the age group, and judging the skin aging degree of the subject according to the comparison result;
or 4) comparing the LRC59 protein content measured in the step 2) with the LRC59 protein content standard curve in the skin of the normally aged person of each age group, and judging the physiological age of the skin of the subject according to the comparison result.
Preferably, the system for assisting in determining the degree of aging comprises the following modules:
(1) a data receiving module; the data receiving module is configured to receive LRC59 protein content data in a skin sample of a subject;
(2) a data storage module: the data storage module is configured to store LRC59 protein content data in normal human skin consistent with the age bracket of the subject;
(3) a data comparison module: the data comparison module is configured to compare the LRC59 protein content data in the skin sample of the subject received by the data receiving module with LRC59 protein content data in normal human skin which is consistent with the age group of the subject and stored in the data storage module;
(4) a judgment module; the judging module is configured to receive the comparison result sent by the data comparing module, judge the comparison result, judge the skin aging degree of the subject, or judge whether the skin physiological age of the subject is consistent with the actual age of the subject, and output the judgment result.
FIG. 1 is a mass spectrum of a characteristic peptide fragment (LQQLPADFGR) of LRC59 protein obtained by detection.
Randomly sampling 7 women and 6 men in normal healthy Chinese as subjects, wherein the relative content data of LRC59 protein in skin samples are as follows:
| group A young group (number) | Age (age) | Relative content of LRC59 protein |
| 1 | 20y (Man) | 14.85221541 |
| 2 | 24y (woman) | 16.25411749 |
| 3 | 25y (Man) | 15.36874363 |
| 4 | 26y (woman) | 14.26189426 |
| 5 | 27y (woman) | 12.43682581 |
| 6 | 31y (Man) | 12.12548934 |
| 7 | 33y (woman) | 11.98426512 |
| Group B youth group (number) | Age (age) | Relative content of LRC59 protein |
| 1 | 55y (Man) | 10.36974673 |
| 2 | 57y (Man) | 11.54134774 |
| 3 | 60y (woman) | 10.26417466 |
| 4 | 63y (woman) | 11.75174679 |
| 5 | 65y (Man) | 9.45845673 |
| 6 | 72y (woman) | 9.62715962 |
As can be seen from the data in the above table, the relative amount of LRC59 protein in the skin samples of subjects decreased with age.
In practical application, firstly, the skin of each statistically significant normal person of each age is collected as a sample, the relative content of the LRC59 protein in each skin sample is respectively measured, for example, to serve the people of 40 years old in a certain city, then firstly, the skin sample of the statistically significant normal person of 40 years old living in the city is collected, the relative content of the LRC59 protein in each skin sample is measured, and the average value is obtained. The average value is a threshold value for measuring the skin aging degree of the subject, when the subject is evaluated, the content of LRC59 protein in the skin is measured by the same method for obtaining the threshold value, and when the content of LRC59 protein is higher than the threshold value, the physiological age of the skin of the subject is younger than the actual age; when the LRC59 protein content in the skin of the subject is lower than the value, the physiological age of the skin of the subject is judged to be older than the actual age.
As to how to measure the content of LRC59 protein in skin, any method capable of determining the absolute and relative content of protein, such as antigen-antibody binding method, etc., other than the method of mass spectrometry in this example, is possible and should be protected by the present invention.
Besides skin, the content of LRC59 protein can also be used as an index for assisting in judging the overall aging degree of human.
Gene:LRC59
Protein LRC59 Protein
MTKAGSKGGNLRDKLDGNELDLSLSDLNEVPVKELAALPKATILDLSCNKLTTLPSDFCGLTHLVKLDLSKNKLQQLPADFGRLVNLQHLDLLNNKLVTLPVSFAQLKNLKWLDLKDNPLDPVLAKVAGDCLDEKQCKQCANKVLQHMKAVQADQERERQRRLEVEREAEKKREAKQRAKEAQERELRKREKAEEKERRRKEYDALKAAKREQEKKPKKEANQAPKSKSGSRPRKPPPRKHTRSWAVLKLLLLLLLFGVAGGLVACRVTELQQQPLCTSVNTIYDNAVQGLRRHEILQWVLQTDSQQ。
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the present invention, and any modifications, equivalents and improvements made within the spirit and principle of the present invention should be included in the scope of the present invention.
Claims (6)
1. A method for non-invasive extraction of LRC59 protein from skin, the method comprising the steps of:
(1) sampling of skin samples of the epidermis of a subject: sticking the 3M medical adhesive patch to the curved side part of the forearm, and slightly removing the 3M adhesive patch after 1 minute to obtain a sticky tape-shaped skin sample;
(2) obtaining of a dried peptide fragment sample: 1) cutting the adhesive tape-shaped skin sample into small pieces, depositing on a glass plate, and transferring to a centrifuge tube;
2) adding a proper amount of lysis buffer sample without SDS, adding 2mM EDTA and 1XCocktail, then placing on ice for 5 minutes, then adding 10mM DTT, and soaking the sample overnight;
3) centrifuging at 25,000g centrifugal force at 4 deg.C for 15 minutes, recovering the supernatant and treating DTT with 10mM for 1 hour in a water bath at 56 deg.C;
4) then treated with 55mM IAM, incubated for 45 minutes at room temperature in the dark, and centrifuged at 25,000g at 4 ℃ for 15 minutes to give the final protein solution supernatant; protein concentration was measured using the Bradford method, and extracted proteins were quality-controlled by 12% SDS-PAGE; taking 100 μ g of protein from each sample, adding trypsin and hydrolyzing at 37 deg.C for 4 hr; then adding trypsin again in the same proportion for enzymolysis for 8 hours at 37 ℃; desalting the polypeptide with Strata X chromatographic column and vacuum drying to obtain dried peptide sample.
2. The method of claim 1, wherein: the method for measuring the relative content of LRC59 protein in an epidermal skin sample based on mass spectrum comprises the following steps: (1) sampling of skin samples of the epidermis of a subject: sticking the 3M medical adhesive patch to the curved side part of the forearm, and slightly removing the 3M adhesive patch after 1 minute to obtain a sticky tape-shaped skin sample;
(2) obtaining of a dried peptide fragment sample: 1) cutting the adhesive tape-shaped skin sample into small pieces, depositing on a glass plate, and transferring to a centrifuge tube;
2) adding a proper amount of lysis buffer sample without SDS, adding 2mM EDTA and 1XCocktail, then placing on ice for 5 minutes, then adding 10mM DTT, and soaking the sample overnight;
3) centrifuging at 25,000g centrifugal force at 4 deg.C for 15 minutes, recovering the supernatant and treating DTT with 10mM for 1 hour in a water bath at 56 deg.C;
4) then treated with 55mM IAM, incubated for 45 minutes at room temperature in the dark, and centrifuged at 25,000g at 4 ℃ for 15 minutes to give the final protein solution supernatant; protein concentration was measured using the Bradford method, and extracted proteins were quality-controlled by 12% SDS-PAGE; taking 100 μ g of protein from each sample, adding trypsin and hydrolyzing at 37 deg.C for 4 hr; then adding trypsin again in the same proportion for enzymolysis for 8 hours at 37 ℃; desalting the polypeptide with Strata X chromatographic column and vacuum drying to obtain dried peptide sample;
(3) detection of
Redissolving the dried peptide fragment sample with mobile phase A (2% ACN, 0.1% FA), centrifuging at 20,000g for 10 min, and sampling the supernatant; separation by ULRC59 protein LC; the sample was first enriched and desalted on a trap column, then connected in series with a self-contained C18 column, at a flow rate of 500nl/min, by the following effective gradient:
separation: 0-5min, 5% mobile phase B (98% ACN, 0.1% FA); 5-160min, mobile phase B increased linearly from 5% to 35%; 160-170min, the mobile phase B rises from 35% to 80%; 170 ℃ 175min, 80% mobile phase B; 176 ℃ for 180min, 5% of mobile phase B; the end of the nanoliter liquid phase separation is directly connected with a mass spectrometer;
DDA mass spectrometric detection
The peptide segment separated by the liquid phase is ionized by a nanoESI source and then is imported to a tandem mass spectrometer Q-active HF mode for detection; setting main parameters: the ion source voltage was set to 1.6 kV; the primary mass spectrum scanning range is 350-1500 m/z; resolution was set to 60,000; the initial m/z of the secondary mass spectrum is fixed to be 100; resolution 15,000. The screening conditions of the parent ions for secondary fragmentation are as follows: parent ions with charges 2+ to 7+, with intensities in excess of 10,000 peak intensity ranked first 20; the ion fragmentation mode is HCD, and fragment ions are detected in Orbitrap; the dynamic exclusion time was set to 30 s; the AGC is set as: primary 3E6, secondary 1E 5;
DIA mass spectrometric detection
The peptide segment separated by the liquid phase is ionized by a nanoESI source and then is imported to a tandem mass spectrometer Q-active HF mode for detection; setting main parameters: the ion source voltage was set to 1.6 kV; the primary mass spectrum scanning range is 350-1500 m/z; resolution was set to 120,000; uniformly dividing 350-1500Da into 40 windows for fragmentation and signal acquisition; the ion fragmentation mode is HCD, and fragment ions are detected in Orbitrap; the dynamic exclusion time was set to 30 s; the AGC is set as: primary 3E6, secondary 1E 5.
3. The method for determining LRC59 protein according to claim 2, wherein: the substance for detecting the content of the LRC59 protein is a mass spectrometric identification reagent, an antibody or an antigen-binding fragment thereof; the substance for detecting the content of the LRC59 protein is an orbital trap high-resolution mass spectrometer.
4. The method for determining LRC59 protein according to claim 2, wherein: the P value of the LRC59 protein is 0.034750959.
5. The LRC59 protein of claim 1, wherein: the method for judging the aging degree and the skin aging degree by the LRC59 protein comprises the following steps:
1) taking a sample of the epidermal skin of a subject;
2) detecting the content of LRC59 protein in the skin sample of the obtained subject;
3) comparing the LRC59 protein content measured in the step 2) with the LRC59 protein content value in the skin of the person with normal aging at the age group, and judging the skin aging degree of the subject according to the comparison result;
or 4) comparing the LRC59 protein content measured in the step 2) with the LRC59 protein content standard curve in the skin of the normally aged person of each age group, and judging the physiological age of the skin of the subject according to the comparison result.
6. The method of claim 5, wherein: the system for assisting in judging the aging degree comprises the following modules:
(1) a data receiving module; the data receiving module is configured to receive LRC59 protein content data in a skin sample of a subject;
(2) a data storage module: the data storage module is configured to store LRC59 protein content data in normal human skin consistent with the age bracket of the subject;
(3) a data comparison module: the data comparison module is configured to compare the LRC59 protein content data in the skin sample of the subject received by the data receiving module with LRC59 protein content data in normal human skin which is consistent with the age group of the subject and stored in the data storage module;
(4) a judgment module; the judging module is configured to receive the comparison result sent by the data comparing module, judge the comparison result, judge the skin aging degree of the subject, or judge whether the skin physiological age of the subject is consistent with the actual age of the subject, and output the judgment result.
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