Diagnostic method for judging brain metabolism waste removal

Abstract

The invention discloses a diagnosis method for judging brain metabolism waste removal. The diagnosis method comprises the following specific steps: S1, sample collection: collecting a biological sample of a subject; step S2, TREM2 and sLRP1 expression level detection: quantitatively detecting the content of TREM2 and sLRP1 in the sample; and S3, data analysis: evaluating the removal efficiency of the brain metabolic waste according to a detection result, specifically, when the expression level of TREM2 or sLRP1 is significantly lower than a baseline level, it can be judged that the removal efficiency of the brain metabolic waste is reduced, and it is prompted that a cognitive disorder risk exists. The invention provides a non-invasive diagnosis method, which can detect the expression levels of TREM2 and sLRP1 at the early stage of neurodegenerative diseases such as cognitive impairment to timely discover the reduction of the removal efficiency of brain metabolic wastes, thereby providing a scientific basis for early intervention.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of biomedical diagnosis, and particularly relates to a diagnostic method for judging brain metabolic waste removal. BACKGROUND

[0002] Circadian rhythm is a key regulatory system for maintaining physiological balance in living organisms, and has an important influence on cognitive function. In recent years, research has found that circadian rhythm disorders may lead to reduced efficiency of brain metabolic waste removal, and thus trigger cognitive dysfunction. TREM2 (triggering receptor expressed on myeloid cells 2) and sLRP1 (triggering receptor expressed on myeloid cells 2) are key components of the endogenous brain removal system, and play an important role in removing abnormal proteins (such as Aβ and Tau protein) in the brain.

[0003] However, there is currently a lack of an effective diagnostic method for judging the brain metabolic waste removal capacity based on the expression level of TREM2 / sLRP1. SUMMARY

[0004] The present application aims to provide a diagnostic method for judging the efficiency of brain metabolic waste removal based on the expression levels of TREM2 and sLRP1, by quantitatively detecting the contents of TREM2 and sLRP1 in biological samples, to evaluate the removal capacity of brain metabolic waste in individuals, and to provide a scientific basis for early diagnosis and intervention of neurodegenerative diseases such as cognitive impairment.

[0005] In order to achieve the above-mentioned purpose, the present application adopts the following technical solutions: A diagnostic method for judging brain metabolic waste removal, comprising the following specific steps: Step S1, sample collection: collecting a biological sample of a subject; Step S2, detection of TREM2 and sLRP1 expression levels: quantitatively detecting the contents of TREM2 and sLRP1 in the sample; Step S3, data analysis: evaluating the removal efficiency of brain metabolic waste according to the detection results, specifically: When the expression level of TREM2 or sLRP1 is significantly lower than the baseline level, it can be judged that the removal efficiency of brain metabolic waste is reduced, indicating a risk of cognitive impairment.

[0006] As a further improvement of the present technical solution: in the step S1, the biological sample is at least one of blood, cerebrospinal fluid or saliva.

[0007] As a further improvement of the present technical solution: in the step S2, the method for quantitatively detecting the contents of TREM2 and sLRP1 is at least one of enzyme-linked immunosorbent assay, Western blotting or real-time fluorescent quantitative PCR.

[0008] As a further improved scheme of the technical solution: further comprising the following steps: establishing a baseline database of TREM2 and sLRP1 expression levels; comparing the differences between the TREM2 and sLRP1 expression levels in the samples of the subjects and the baseline levels to evaluate the brain metabolic waste removal efficiency.

[0009] As a further improved scheme of the technical solution: the establishment of the baseline database includes collecting biological samples of healthy people at different stages of circadian rhythm, detecting and recording the expression levels of TREM2 and sLRP1.

[0010] As a further improved scheme of the technical solution: further including combining the clinical information of the subjects, including at least one of age, gender, sleep quality, to further analyze the factors affecting the expression levels of TREM2 and sLRP1.

[0011] As a further improved scheme of the technical solution: the diagnostic method is used for early screening of cognitive impairment, and the changes of brain metabolic waste removal efficiency are monitored by regularly detecting the expression levels of TREM2 and sLRP1.

[0012] As a further improved scheme of the technical solution: further including generating a diagnostic report, the report content including the detection results of the expression levels of TREM2 and sLRP1, the evaluation of the brain metabolic waste removal efficiency and the corresponding health suggestions.

[0013] The application also provides the use of the diagnostic method according to any one of the above in the preparation of a cognitive impairment diagnostic kit or device, which comprises reagents for detecting the expression levels of TREM2 and sLRP1 and data analysis software.

[0014] Compared with the prior art, the beneficial effects of the present application are: 1、The present application provides a non-invasive diagnostic method, which can detect the expression levels of TREM2 and sLRP1 at the early stage of cognitive impairment and other neurodegenerative diseases, and timely find the decrease of brain metabolic waste removal efficiency, providing a scientific basis for early intervention.

[0015] 2、Combined with the clinical information of the subjects, the present application can provide personalized health suggestions and treatment plans for different individuals, and improve the treatment effect.

[0016] 3、The present application provides a new research tool and method for studying the relationship between circadian rhythm and brain metabolic waste removal, which helps to further explore the pathogenesis of neurodegenerative diseases.

[0017] The above description is only a summary of the technical solutions of the present application. In order to make the technical means of the present application clearer and can be implemented according to the content of the specification, the following will be described in detail with the preferred embodiments of the present application and the accompanying drawings. The specific embodiments of the present application are given in detail by the following examples and the accompanying drawings. BRIEF DESCRIPTION OF DRAWINGS

[0018] The accompanying drawings, which are included to provide a further understanding of the present application, constitute a part of this application and illustrate illustrative embodiments of the present application and its description serve to explain the present application, and do not constitute an improper limitation on the present application. In the drawings: Figure 1 A flowchart of a diagnostic method for judging brain metabolic waste removal according to the present application. DETAILED DESCRIPTION

[0019] The principles and features of the present application will be described below in conjunction with the accompanying drawings. The examples are only used to explain the present application and are not intended to limit the scope of the present application. In the following paragraphs, the present application is described in more detail by way of example with reference to the accompanying drawings. It should be noted that the drawings are greatly simplified and use non-precise proportions, only to facilitate and clearly assist in explaining the purpose of the embodiments of the present application.

[0020] Referring to Figure 1 The present application proposes a diagnostic method for judging brain metabolic waste removal, comprising the following specific steps: Step S1, sample collection: Collect biological samples of the subjects, including but not limited to blood, cerebrospinal fluid or saliva, etc.

[0021] The sample collection time points should cover different stages of circadian rhythm to comprehensively evaluate the influence of circadian rhythm on the expression of TREM2 and sLRP1.

[0022] Step S2, detection of TREM2 and sLRP1 expression levels: Quantitative detection of the content of TREM2 and sLRP1 in the sample by enzyme-linked immunosorbent assay (ELISA), Western Blot (WB) or real-time fluorescent quantitative PCR (qPCR) method, etc.

[0023] The specific operation steps are carried out according to the instructions of each kit or device to ensure the accuracy and reliability of the detection results.

[0024] Step S3, data analysis: Collect and organize the detection data to establish a baseline database of the expression levels of TREM2 and sLRP1.

[0025] By comparing the expression levels of TREM2 and sLRP1 at different time points and different circadian rhythm states, the clearance efficiency of brain metabolic waste is evaluated.

[0026] In combination with the clinical information of the subject (such as age, gender, sleep quality, etc.), further analysis of the factors affecting the expression levels of TREM2 and sLRP1 is performed.

[0027] Step S4, diagnosis criteria: According to the detection results of the expression levels of TREM2 and sLRP1, in combination with the clinical information, the diagnosis criteria of the clearance efficiency of brain metabolic waste is established.

[0028] For example, when the expression level of TREM2 or sLRP1 is significantly lower than the baseline level, it can be judged that the clearance efficiency of brain metabolic waste is reduced, which indicates the risk of cognitive impairment; When the expression level of TREM2 or sLRP1 is within the baseline level range or significantly higher than the baseline level, it can be judged that the subject has normal brain metabolic waste clearance ability or compensatory activation.

[0029] Step S5, diagnosis report generation: According to the data analysis results, a detailed diagnosis report is generated, including the detection results of the expression levels of TREM2 and sLRP1, the evaluation of the clearance efficiency of brain metabolic waste, and the corresponding health recommendations.

[0030] Example 1: Detection of TREM2 and sLRP1 expression levels in blood samples and their association with cognitive impairment 1.1 Experimental subjects Healthy volunteer group: 30, aged 20-40 years old, no history of cognitive impairment, PSQI score ≤5.

[0031] Cognitive impairment patient group: 30, aged 50-70 years old, diagnosed with mild cognitive impairment (MCI) or Alzheimer's disease (AD), PSQI score ≤5.

[0032] 1.2 Sample collection 5mL of fasting venous blood was collected from the two groups of subjects in the morning, and the serum was separated by centrifugation and stored at -80℃ for standby.

[0033] 1.3 Experimental method The concentrations of TREM2 and sLRP1 in serum were detected using ELISA kits (Human TREM2 ELISA Kit, Human sLRP1 ELISA Kit).

[0034] The operation steps are strictly carried out according to the kit instructions, including standard preparation, sample dilution, sample addition, incubation, washing, color development and termination reaction, etc.

[0035] The optical density (OD value) of each well was measured at 450 nm wavelength using an enzyme label meter, and the concentrations of TREM2 and sLRP1 were calculated according to the standard curve.

[0036] 1.4 Experimental results Healthy volunteer group: the average concentration of TREM2 was 275.6 ± 32.1 pg / mL, and the average concentration of sLRP1 was 30.2 ± 4.5 μg / mL.

[0037] Cognitive impairment patient group: the average concentration of TREM2 was 198.4 ± 28.7 pg / mL, which was significantly lower than that of the healthy volunteer group (P < 0.01); the average concentration of sLRP1 was 22.1 ± 3.8 μg / mL, which was also significantly lower than that of the healthy volunteer group (P < 0.01).

[0038] 1.5 Data analysis and conclusion The expression levels of TREM2 and sLRP1 in the serum of cognitive impairment patients were significantly lower than those of healthy volunteers, indicating that the clearance efficiency of brain metabolic waste was reduced.

[0039] The present application can effectively evaluate the clearance ability of brain metabolic waste of individuals by detecting the concentrations of TREM2 and sLRP1 in the serum, and provide a scientific basis for early diagnosis of cognitive impairment.

[0040] Example Two: Effect of Circadian Rhythm on Expression Levels of TREM2 and sLRP1 2.1 Experimental subjects The same batch of healthy volunteers (n = 30) were collected blood samples under normal and reversed circadian rhythm conditions.

[0041] 2.2 Experimental design Normal circadian rhythm group: the subjects maintained regular sleep-wake cycles (bedtime at 22:00, wake-up time at 06:00), and blood samples were collected at 8:00 the next morning.

[0042] Reversed circadian rhythm group: the subjects experienced 3 days of sleep deprivation (maintained wakefulness from 22:00 to 06:00), and blood samples were collected at 8:00 the next morning.

[0043] 2.3 Sample collection and detection The same as examples 1.2 and 1.3.

[0044] 2.4 Experimental results Normal circadian rhythm group: the average concentration of TREM2 was 275.6 ± 32.1 pg / mL, and the average concentration of sLRP1 was 30.2 ± 4.5 μg / mL.

[0045] The average concentration of TREM2 was 210.3 ± 29.6 pg / mL, which was significantly lower than that of the normal circadian rhythm group (P < 0.05); the average concentration of sLRP1 was 25.7 ± 4.1 μg / mL, which was also significantly lower than that of the normal circadian rhythm group (P < 0.05).

[0046] 2.5 Data analysis and conclusion The circadian rhythm reversal significantly reduced the expression levels of TREM2 and sLRP1 in serum, suggesting that the circadian rhythm disorder may affect the clearance efficiency of brain metabolic waste.

[0047] The present application can evaluate the influence of the circadian rhythm on the clearance efficiency of brain metabolic waste by detecting the concentration changes of TREM2 and sLRP1 under different circadian rhythm states, thereby providing a new method for early screening of circadian rhythm-related cognitive impairment.

[0048] The above is only a preferred embodiment of the present application, and does not limit the present application in any form; any person skilled in the art can easily implement the present application according to the drawings and the above description; however, any equivalent changes, modifications and evolutions made by those skilled in the art within the scope of the technical solutions of the present application, using the disclosed technical content, are equivalent embodiments of the present application; at the same time, any equivalent changes, modifications and evolutions made according to the essence of the present application to the above embodiments are still within the protection scope of the technical solutions of the present application.

Claims

1. A diagnostic method for assessing the clearance of metabolic waste products in the brain, characterized in that, The specific steps include the following: Step S1, Sample Collection: Collect biological samples from the subject; Step S2, TREM2 and sLRP1 expression level detection: Quantitatively detect the content of TREM2 and sLRP1 in the sample; Step S3, Data Analysis: Evaluate the brain metabolic waste removal efficiency based on the test results, specifically: When the expression levels of TREM2 or sLRP1 are significantly lower than baseline levels, it can be judged as a decrease in the efficiency of brain metabolic waste clearance, suggesting a risk of cognitive impairment.

2. The diagnostic method for determining the clearance of brain metabolic waste according to claim 1, characterized in that, In step S1, the biological sample is at least one of blood, cerebrospinal fluid, or saliva.

3. The diagnostic method for determining the clearance of brain metabolic waste according to claim 2, characterized in that, In step S2, the method for quantitatively detecting the content of TREM2 and sLRP1 is at least one of enzyme-linked immunosorbent assay (ELISA), Western blotting, or real-time quantitative PCR.

4. The diagnostic method for determining the clearance of brain metabolic waste according to claim 3, characterized in that, It also includes the following steps: Establish a baseline database of TREM2 and sLRP1 expression levels; compare the differences between TREM2 and sLRP1 expression levels in the subject samples and the baseline levels to assess the efficiency of brain metabolic waste clearance.

5. The diagnostic method for determining the clearance of brain metabolic waste according to claim 4, characterized in that, The establishment of the baseline database involves collecting biological samples from healthy individuals at different stages of their circadian rhythm, and detecting and recording the expression levels of TREM2 and sLRP1.

6. The diagnostic method for determining the clearance of brain metabolic waste according to claim 5, characterized in that, It also includes combining the subjects' clinical information, including at least one of age, sex, and sleep quality, to further analyze the factors affecting the expression levels of TREM2 and sLRP1.

7. The diagnostic method for determining the clearance of brain metabolic waste according to claim 6, characterized in that, The diagnostic method is used for early screening of cognitive impairment by periodically detecting the expression levels of TREM2 and sLRP1 to monitor changes in the efficiency of brain metabolic waste clearance.

8. The diagnostic method for determining the clearance of brain metabolic waste according to claim 7, characterized in that, It also includes generating diagnostic reports, which include the results of TREM2 and sLRP1 expression level detection, assessment of brain metabolic waste clearance efficiency, and corresponding health recommendations.

9. The application of the diagnostic method according to any one of claims 1 to 8 in the preparation of a diagnostic kit or device for cognitive impairment, characterized in that, The kit or device contains reagents and data analysis software for detecting the expression levels of TREM2 and sLRP1.

Images