CN115192716A

CN115192716A - Methods and systems for predicting, preventing or treating post-operative delirium

Info

Publication number: CN115192716A
Application number: CN202210950424.9A
Authority: CN
Inventors: 张红超; 陈霞; 卫小娟; 张致同; 张济远
Original assignee: Air Force Specialty Medical Center of PLA
Current assignee: Air Force Specialty Medical Center of PLA
Priority date: 2022-08-09
Filing date: 2022-08-09
Publication date: 2022-10-18

Abstract

The present invention discloses a method and system for predicting, preventing or treating post-operative delirium. The prediction method comprises the following steps: a step of measuring the content of PGC-1 α and/or indolpropanoic acid in a sample collected from the subject using an agent to obtain a measured value; a step of comparing the measured value with a standard value; when the measured values are below the normative values, the subject is detected as having, or predicted to be at risk for having, post-operative delirium. The invention researches the molecular mechanism of development of the delirium after the operation of the human body, provides a new mechanism insight for the pathogenesis of the delirium after the operation, and further promotes to establish an effective prevention strategy.

Description

Methods and systems for predicting, preventing or treating post-operative delirium

Technical Field

The present invention relates to the field of biomedicine, and in particular to methods and systems for predicting, preventing or treating post-operative delirium.

Background

The incidence of postoperative delirium (POD) is about 9-50% in patients undergoing anesthesia and surgery. The occurrence of POD is associated with a higher incidence of postoperative complications, poorer clinical outcome, and higher mortality of the patient. The pathogenesis of POD is still largely unclear at present. Age, complex complications, preoperative cognitive insufficiency, sleep deprivation, type of surgery, and the like are considered to be important risk factors for POD. In particular, the higher the age, the greater the probability of occurrence of POD. With the aging population and the increasing need for anesthesia and surgery, POD has become an urgent issue related to public health.

To date, there is still a clinical lack of reliable blood biomarkers and effective prophylactic and therapeutic drugs for POD.

Disclosure of Invention

In order to solve at least some of the technical problems of the prior art, the present invention has conducted metabolomics studies on prospective patient cohorts and determined key factors associated with POD pathogenesis. The invention further tests the potential of the molecule for preventing and treating POD in preclinical animal models, and examines the action mechanism of the molecule. Specifically, the present invention includes the following.

In a first aspect of the present invention, there is provided a use of PGC-1 α or a production promoter thereof for the preparation of a medicament for the prevention and/or treatment of postoperative delirium.

In certain embodiments, the use according to the invention, wherein the prevention and/or treatment refers to curing, alleviation, mitigation, prevention of occurrence and development of post-operative delirium or post-operative delirium-like behavior.

In certain embodiments, the use according to the present invention, wherein the promoter of PGC-1 α production comprises a tryptophan-related metabolite of the gut microflora, wherein said metabolite comprises indole propionic acid or an analogue thereof.

In a second aspect of the invention, there is provided the use of PGC-1 α and/or indolpropion in the manufacture of an agent for predicting the risk of post-operative delirium occurrence.

In certain embodiments, the use according to the invention, wherein the predicting comprises:

(1) A step of measuring the content of PGC-1 α and/or indolpropanic acid in a sample collected from a subject using an agent to obtain a measured value;

(2) A step of comparing the measured value with a standard value;

(3) When the measurement values are below the normative values, then detecting the subject as having, or predicting the subject as being at risk for having, post-operative delirium; when the measurement value is higher than the normative value, then the subject is detected as not suffering from post-operative delirium, or the subject is predicted as not being at risk of suffering from post-operative delirium.

In certain embodiments, the use according to the invention, wherein the standard value is a value obtained from a biological sample of a normal subject comparable to the age of the subject, or the standard value is a value obtained from a biological sample of the same subject prior to surgery.

In certain embodiments, the use according to the invention, wherein the amount in the sample T1 taken from the subject at a first point in time is measured as the standard value and the amount of the product T2 taken from the same subject at a second point in time is measured as the measured value.

In a third aspect of the invention, there is provided a method of screening for a compound useful for the treatment of post-operative delirium comprising:

a. a step of measuring the content of PGC-1 α and/or indolpropionic acid in a sample collected pre/post-operatively from a subject suffering from post-operative delirium or post-operative delirium-like behavior.

b. A step of administering the compound to the subject;

c. a step of measuring the content of PGC-1 α and/or indolpropanoic acid in a sample collected from the subject following administration of the compound to yield a second measurement value;

d. a step of comparing the first measurement value and the second measurement value;

e. selecting the compound as a compound not useful for treating or slowing postoperative delirium or postoperative delirium-like behavior when the second measurement is less than or equal to the first measurement, and selecting the compound as a compound useful for treating or slowing postoperative delirium or postoperative delirium-like behavior when the second measurement is greater than the first measurement.

In a fourth aspect of the invention, there is provided a system for predicting risk of onset of post-operative delirium, comprising:

a data acquisition unit for acquiring at least PGC-1 α and/or indole propionic acid content data from a subject; and

a determination unit for classifying the subject as belonging to a patient with a high or low risk of developing postoperative delirium based on the comparison of the content of PGC-1 alpha and/or indolpropion with a predetermined threshold.

In a fifth aspect of the invention, there is provided a method for the prevention and/or treatment of post-operative delirium comprising administering to a subject in need thereof a therapeutically effective amount of PGC-1 α and/or indolpropionic acid.

In a sixth aspect of the invention, there is provided a method for detecting post-operative delirium comprising an agent that indicates the amount of PGC-1 α and/or indole propionic acid.

According to the invention, through research, the intestinal microbiota metabolite indole propionic acid is found to be negatively related to the occurrence of human POD. In preclinical animal models for POD assessment, lack of indolpropanic acid promotes POD-like behavior, whereas exogenous indolpropanic acid administration prevents POD-like behavior. This protective effect of indole propionic acid is mediated in part by PGC-1 α and interneurons in the hippocampus. These results provide new mechanistic insights into POD pathogenesis in order to develop further preventive strategies.

Drawings

FIG. 1 is a graph showing the results of analyzing the concentration of indolpropion in plasma samples from patients before and after surgery using LC-MS/MS techniques.

Figure 2 shows the behavioral score and relative abundance of indolpropane results for normal control and sterile mouse models.

Figure 3 shows that in vivo supplementation with indolpropionic acid reversed the occurrence of POD after anesthesia in mice.

FIG. 4 shows the results of detecting the levels of PGC-1. Alpha. In hippocampal tissue of ampicillin-treated/untreated mouse brain and the expression of PGC-1. Alpha. Induced by IPA using western blot technique.

FIG. 5 shows that increasing PGC-1. Alpha. Protein in hippocampal brain reduces POD occurrence.

Detailed Description

Reference will now be made in detail to various exemplary embodiments of the invention, the detailed description should not be construed as limiting the invention but as a more detailed description of certain aspects, features and embodiments of the invention.

It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. Further, for numerical ranges in this disclosure, it is understood that the upper and lower limits of the range, and each intervening value therebetween, is specifically disclosed. Every smaller range between any stated value or intervening value in a stated range and any other stated or intervening value in a stated range is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference herein for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control. Unless otherwise indicated, "%" is percent by weight.

The present invention is described in detail below.

As used herein, the terms "post-operative delirium", "post-operative delirium" and "POD" are used interchangeably and include disorders of consciousness, cognition, orientation, thinking, memory and sleep that occur post-operatively, e.g., post-anesthesia surgery, which is a fluctuating acute psychotic disorder syndrome, sometimes also referred to as post-operative psychotic disorder or post-operative cognitive dysfunction.

The present invention provides the use of PGC-1 α or a production promoter thereof for the preparation of a medicament for the prevention and/or treatment of postoperative delirium. PGC-1 α refers to peroxisome proliferator activated receptor- γ coactivator-1 α, the production facilitator including, but not limited to, a compound, agent or formulation capable of facilitating, stimulating, activating, increasing, enhancing PGC-1 α production, which includes a tryptophan-related metabolite of the gut microflora, preferably wherein said metabolite comprises Indole Propionic Acid (IPA) or an analog thereof.

The term "analog" as used herein refers to a compound that is structurally related to the parent compound and contains different functional groups or substituents. For example, where the parent compound of the present invention comprises indole propionic acid, the chemical analogs of indole propionic acid will contain certain functional groups and substituents that are different from those of indole propionic acid.

Production of PGC-1 α and/or indole propionic acid may be manifested in altered levels thereof, such as an increase or decrease in the amount of PGC-1 α and/or indole propionic acid. It is understood that their activity/function is related to their physical quantities. In some embodiments, PGC-1 α levels refer to protein expression levels, or mRNA expression levels of PGC-1 α -related genes.

In the present invention, the intestinal microbial flora refers to those microorganisms capable of metabolizing tryptophan, and preferably also to those microorganisms which produce metabolites of at least trimethylamine oxide, indole propionic acid or the like by tryptophan metabolism. In certain embodiments, the member of the gut microflora that produces indole propionic acid is associated with the fldABC gene family. In certain embodiments, the gut microbial flora of the present invention includes clostridium sporogenes and streptococcus digestans.

The invention further provides the use of PGC-1 a and/or indolpropanic acid in the manufacture of an agent for predicting the risk of developing postoperative delirium, said prediction comprising at least:

(1) A step of measuring the content of PGC-1 α and/or indolpropanoic acid in a sample collected from the subject using an agent to obtain a measured value;

(2) A step of comparing the measured value with a standard value;

(3) When the measurement values are below the normative values, then detecting the subject as having, or predicting the subject as being at risk for having, post-operative delirium; when the measured value is higher than the normative value, then the subject is detected as not suffering from, or predicted as not being at risk of suffering from, postoperative delirium.

Step (1)

Step (1) of the present invention is a step for obtaining a measured value of the content of PGC-1. Alpha. And/or indole propionic acid in a sample of a subject. Wherein the type of sample is not limited, examples of which include, but are not limited to, tissue samples or fluid samples. Tissue samples include somatic cell samples, and fluid samples include blood or components thereof such as plasma, serum, and the like. The biological sample may be any sample of mammalian origin, preferably of human origin. Examples of types of biological samples that may be used in the present invention include, but are not limited to, one or more of the following: urine, feces, tears, whole blood, serum, plasma, blood components, bone marrow, cells, tissues, organs, body fluids, saliva, cheek swabs, lymph fluid, cerebrospinal fluid, lesion exudate, and other fluids produced by the body. The biological sample may also be a frozen, fixed, paraffin embedded or fresh biopsy sample.

In the present invention, the reagent refers to any reagent that can be used to indicate the content or level of PGC-1 alpha and/or indolpropane to be tested. Preferably, the agent includes, but is not limited to, an antibody. Antibodies herein specifically encompass monoclonal antibodies (including full length monoclonal antibodies), polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments, such as variable domains and other portions of antibodies that exhibit the desired biological activity. The term "monoclonal antibody (mAb)" refers to an antibody that is highly specific and directed against a single antigenic determinant (epitope). Thus, the term "monoclonal" refers to an antibody to the same epitope and should not be construed as requiring the production of the antibody by any particular method. It is understood that monoclonal antibodies can be prepared by any technique or method known in the art; including, for example, the hybridoma method (Kohler et al, 1975, nature256: clackson et al, 1991, nature 352; and Marks et al, 1991, J.mol.biol.222.

In the present invention, an "antibody fragment" refers to a molecule that is different from an intact antibody, comprises a portion of an intact antibody, and binds to an antigen to which the intact antibody binds. Examples of antibody fragments include, but are not limited to, fv, fab '-SH, F (ab') 2; a diabody; a linear antibody; single chain antibody molecules (e.g., scFv); and multispecific antibodies formed from antibody fragments.

The agent of the present invention may comprise other components in addition to the above-mentioned antibody. Examples of other components include, but are not limited to, LC-MS/MS related assay reagents. In certain embodiments, any one of the above substances may be separately present in a state separated from the other substances, and stored in different containers (e.g., vials), as long as they are capable of contacting each other at the time of use. In addition, preferably, any two or more of the above-described substances may be mixed to exist as a mixture.

In certain embodiments, the other ingredients may be provided separately in the form of a dry powder, or in the form of a solution, e.g., in the form of an aqueous solution. The concentrations or contents of these substances, in the case of their presence in aqueous solution, are readily determinable by the person skilled in the art as a function of the various requirements. For example, for storage purposes, the concentration of the substance may be present in a higher form, and when in service or in use, the concentration may be reduced to a working concentration by, for example, diluting the higher concentration solution.

The agents of the invention can further be prepared as diagnostic agents for detecting post-operative delirium in a subject. The diagnostic agent can be in the form of a diagnostic composition, a diagnostic kit, or any other form in which a plurality of separately present reagents are used in combination.

In the present invention, PGC-1. Alpha. And its fragments are preferably detected by known means for quantitative detection of proteins. Quantitative detection of proteins is known in the art, e.g., measurement of measured values and/or standard values can be performed using immunoblotting techniques, LC-MS/MS protein quantification techniques, and the like.

Step (2)

Step (2) of the present invention is a step of comparing the measured value with a standard value. The standard value may be a specific value or a range of values.

In certain embodiments, a standard value can be a sample test value from a normal subject. Preferably, the values of the sample from a normal subject of an age comparable to that of the subject to be tested, and also preferably, the standard values and the measured values are obtained by the same method.

In certain embodiments, the standard values are from different time periods of the same subject. For example, the amount in the sample T1 taken from the subject at a first point in time is measured as the standard value and the amount in the sample T2 taken from the same subject at a second point in time is measured as the measured value. Wherein the first time point T1 is preferably a time point before no indication of postoperative delirium is shown or present and the time point at which the second time point T2 is detected is preferably a time point after surgery, e.g. after performing an anesthesia operation.

Step (3)

Step (3) of the present invention is a result determination step. In particular, when said measured values are below said normative values, then said subject is detected as having, or predicted to be at risk of having, post-operative delirium; when the measurement value is higher than the normative value, then the subject is detected as not suffering from post-operative delirium, or the subject is predicted as not being at risk of suffering from post-operative delirium.

The invention also provides a method, sometimes referred to herein simply as a "screening method", for screening for compounds useful in treating or alleviating post-operative delirium. Preferably, the method comprises:

a. a step of measuring the content of PGC-1 α and/or indolpropanoic acid in a sample collected pre/post-operatively from a subject suffering from post-operative delirium or post-operative delirium-like behavior resulting in a first measurement;

b. a step of administering said compound to a subject suffering from post-operative delirium or post-operative delirium-like behavior;

c. a step of measuring the content of PGC-1 α and/or indolpropionic acid in a sample collected from a subject suffering from post-operative delirium or post-operative delirium-like behavior after administration of said compound to obtain a second measurement value;

In the screening method of the present invention, the subject is preferably an animal model, for example, rat, mouse, dog, pig, monkey, orangutan, or the like, which has postoperative delirium or postoperative delirium-like behavior. Such animals can be artificially induced to suffer from post-operative delirium or post-operative delirium-like behavior.

The invention further provides a system for predicting the risk of occurrence of postoperative delirium comprising:

a determination unit for classifying the subject as belonging to a patient at high or low risk of developing postoperative delirium based on a comparison of the PGC-1 α and/or indolpropanoic acid content with a predetermined threshold.

The data of the data acquisition unit can be data obtained by hospital laboratory examination, and can also be acquired by various possible acquisition ways, such as a blood detection instrument, a detection device based on a protein detection method or mass spectrometry, liquid chromatography-mass spectrometry and the like and having certain data processing and storage capacity. Such detection devices include, but are not limited to, an immunoassay analyzer, a liquid chromatography mass spectrometry system or platform, and the like.

The detection/identification value of the system or method of the invention can be determined by, for example, calculating the evaluation indicators such as the area under the working characteristic curve (AUC), sensitivity, specificity, etc. of the subject. Where AUC is defined as the area under the ROC curve bounded by coordinate axes, said area having a value in the range between 0.5 and 1. The closer the AUC is to 1.0, the higher the authenticity of the assay.

The present invention further provides a kit for the detection of post-operative delirium in a subject comprising reagents for measuring the content of PGC-1 α and/or indolpropion in a sample taken from said subject to obtain a measurement. Kits of the invention may further comprise other reagents capable of routine detection by various assay types, such as ELISA assays, immunoassays, protein chips or microarrays, DNA/RNA chips or microarrays, RT-PCR, second or third generation sequencing, mass spectrometry, immunohistochemistry, flow cytometry, or high content cell screening.

It will be appreciated by those skilled in the art that other steps or operations, before, after, or between the steps of the methods involved, may be included as long as the objects of the invention are achieved, for example to further optimize and/or improve the methods described herein.

Examples

1. Materials and methods

1. And (3) mice: all procedures and animal use were approved by the Institutional Animal Care and Use Committee (IACUC) and were in compliance with guidelines established by the Chinese and International society for pain research. Female C57BL/6 mice were purchased from the laboratory animals center of the military medical academy of sciences. Mice were used at 4-6 months of age. For the perturbation of the intestinal microbiota, the mice were provided with 0.5g/L ampicillin in freely provided drinking water.

2. Cell culture and treatment: the mouse hippocampal HT-22 cell line used in the study was purchased from the laboratory animal center of the academy of military medical sciences and maintained in Dulbecco's Modified Eagle Medium (DMEM)/high glucose (Gibco, thermo Fisher Scientific, waltham, MA, USA) supplemented with 10% Fetal Bovine Serum (FBS) and 100U/ml penicillin/streptomycin (Gibco). The culture was maintained at 37 ℃ and 5% CO humidified ₂ An incubator. Subsequent experiments were performed when the cultured cells were 80-85% confluent. IPA was purchased from Sigma Aldrich (MO, USA), 10mM stock was prepared and stored at-80 deg.C until use. Immediately prior to the experiment, aliquots of IPA were dissolved and diluted to the final concentration in the culture medium. 5, 10, 50 and 100uM IPA was applied to the cells for 6 hours. The control cells were cultured in normal DMEM medium under normoxic conditions at 37 ℃. Cells were harvested for subsequent experiments.

3. Human subjects: the human study was approved by the review board of the national release force air force feature medical center, and all subjects signed an informed consent to participate in the study. Blood samples were collected prior to anesthesia and surgery. Within 36 hours after surgery, the subjects were scored by two different neurologists according to the internationally accepted diagnostic criteria of the mental assessment method (CAM), and the evaluation criteria of the two neurologists were combined to determine whether the patients suffered from postoperative delirium.

4. POD judgment of mouse animal model: delirium-like phenotype after anesthesia/surgery in mice was determined by using an internationally recognized and suggested series of tests. 6 hours after the operation of the tested mouse, the mouse is subjected to standard behavior tests such as 1) novelty identification test, 2) buried food test, 3) Y maze test, 4) open field test and the like, and finally, the Z score value is calculated after comprehensive scoring to judge whether the animal suffers from postoperative delirium (POD).

5. Anesthesia and surgery of mice: mice were anesthetized with an isoflurane vaporizer. Lidocaine 1% pure was used for skin infiltration. Surgery was performed on the bilateral thighs to expose the femoral artery, taking care not to dissect the femoral/sciatic nerve. Each side of the skin incision was about 0.6cm; the wound was closed with 2-0 silk suture. For sham surgery, animals received isoflurane anesthesia for a similar time, with 1% lidocaine used for skin infiltration.

6. Metabonomics based on high performance liquid chromatography-mass spectrometry tandem (LC-MS/MS) technology: 70. Mu.l of acetonitrile/methanol solution (1,v/v) was added to a centrifuge tube containing 30. Mu.l of plasma samples of human subjects and animals, and after shaking and mixing, high molecular components such as proteins were precipitated, and after low-temperature high-speed centrifugation at 14,000rpm, the supernatant fraction was separated and transferred. Mu.l of the supernatant was automatically applied to an XBridge Amide (waters, USA) chromatographic column for HPLC separation, and the separated components were directly introduced into a tandem mass spectrometer (QQQ-6490, agilent, USA) for qualitative and semi-quantitative detection of the targeted metabolic molecules. Mass spectral data were subjected to peak extraction and quantitative analysis using MassHunter quantitative analysis software from Agilent. The metabonomic platform technology can be used for detecting more than 350 substances such as biological acid, nucleotide, carbohydrate metabolite, biological amine, intestinal flora metabolite, bile acid, free fatty acid, fatty acylated carnitine and the like in a targeted manner.

7. Protein extraction and western blotting: cultured cells were lysed in RIPA buffer (Cell Signaling, USA) containing protease inhibitors (thermoldisser, USA). Frozen tissue was minced while still slightly frozen and homogenized in lysis buffer as described above. The samples were then centrifuged at 21,000 Xg for 10 minutes at 4 ℃. Supernatants were collected and protein quantification was performed using the Bio-Rad protein assay (Biorad, mississauga, ontario). For immunoblotting, protein samples were separated on 10% SDS-PAGE for 2 hours at 100V constant voltage. The proteins dissolved in the gel were then transferred to a PVDF membrane. Staining was performed using the following antibodies: anti-PGC 1 α (1, 1000, rabbit polyclonal; abcam), or anti- β -actin (1.

8. Statistical analysis: metabonomics analysis of a sample tested by a crowd is carried out by adopting a logistic regression analysis method of an SAS software package; data analysis of the test animals was performed using a two-tailed t-test.

2. Results of the experiment

Human subjects undergoing bone surgery were included in the clinical study of POD. As part of the study, venous blood samples were obtained from each subject prior to surgery. These subjects were then followed up for 48 hours post-surgery to monitor POD development. Diagnosis of POD was performed using a conscious fuzzy evaluation method (CAM), and blood samples were analyzed using liquid chromatography and tandem mass spectrometry by an analyst blinded to the study design. For a total of 98 subjects whose blood samples were available for metabolomics studies, 9 subjects developed POD, while the remaining 86 did not. There were no statistical differences in age, gender, and postoperative pain scores between the two groups when comparing demographics of subjects with POD (POD group) and without POD (non-POD group). Of the approximately 350 molecules of the semi-quantitative analysis by metabolomics, we found that there were significant differences in the amount of multiple molecules in the blood samples from the two groups of patients (results not shown). Of these, only the intestinal flora metabolite indole-3-propionic acid (IPA), was the only one that was significantly reduced in preoperative blood samples with post-operative POD (fig. 1). Simple logistic analysis showed that IPA levels in plasma were negatively correlated with the occurrence of post-anesthesia delirium in patients (β = -2.55, p-val = 0.01).

The present invention further examined whether perturbation of the gut microbiota could affect the development of POD. For this purpose, mice were supplemented with ampicillin-containing water (Amp group) or plain water (H) ₂ Group O), then the two groups of animals were subjected to anesthesia surgery and sham surgery, and the animals were evaluated for behavior at 6 hours after the surgery to determine whether POD was produced; plasma samples from the test animals were also collected for metabonomics analysis. Cognitive dysfunction score determinations were performed 6 hours after surgery or anesthesia/surgery (a/S) and the results showed no statistically significant differences in the cognitive dysfunction scores of the test animals in the sham operated animal test population, whether given normal food water retention gut microbes or ampicillin clearance gut microbes (p =0.66, two tail t test); however, in the group of animals tested given anesthesia and surgery, there was a statistically significant difference in cognitive function scores between the two groups of animals fed water and ampicillin (p =4.08E-7, double global t-test) (fig. 2 left). Data are represented as Sham + H ₂ Group O, group Sham + Amp, group A/S+H ₂ Mean ± SE of O or a/S + Amp groups (n =12 animals per group). To determine whether the IPA concentration in the test mice was affected by feeding ampicillin, peripheral blood was collected from all the test animals, and the IPA concentration in the plasma of the mice was measured by LC-MS/MS technique. The results showed that IPA concentration in plasma was significantly reduced in the middle east of ampicillin fed animals, both in sham and post-anaesthesia surgical groups (sham p =0.0002, surgical p =0.003, two-tailed t test) (fig. 2 right). Data are expressed as Sham + H ₂ Group O, group Sham + Amp, group A/S + H ₂ Mean ± SE of O or a/S + Amp groups (n =10 animals per group).

In view of the innovative findings that IPA content in the population is negatively correlated with post-operative POD, and that after artificially reducing IPA levels by feeding antibiotics, the test animals suffering from POD are significantly elevated, the present invention further studies exogenous administration of IPA, and observes whether IPA prevents development of mouse POD-like behavior. Using the same technique, the present inventors fed conventional water (H) to mice ₂ O), ampicillin-containing water (Amp), or fed with ampicillin-containing water and intraperitoneal injection of IPA (Amp + IPA), followed by anesthesia/surgery (a/S) treatment (AH). Cognitive dysfunction score measurements were performed 6 hours after a/S (figure 3) and showed a significant increase in cognitive dysfunction scores in the group of mice fed ampicillin (p =1.4E-07, two-tailed t-test), further confirming that deprivation of gut microbes could lead to significant podding in the animal development. However, in the group of test animals supplemented with IPA by intraperitoneal injection, the cognitive dysfunction score was significantly lower than that of the animals not supplemented with IPA (p =5.81E-08, two-tailed t test); cognitive dysfunction scores were not statistically significantly different between the two groups of animals fed regular chow and fed ampicillin-containing animals with intraperitoneal injections of IPA (figure 3).

To determine the likely mechanism by which IPA affects POD development, the present invention tests peroxisome proliferator activated receptor gamma coactivator 1-alpha (PGC-1 α), a major regulator of mitochondrial biogenesis, and plays a key role in neuronal function, including gabaergic interneurons. The inventor firstly adopts the immunoblotting technology to detect the protein content of PGC-1 alpha in hippocampal region of brain of tested animal. As a result, it was found that the protein content of PGC-1. Alpha. In the hippocampal tissue lysate of the ampicillin-fed mice was significantly lower than that of the same brain region of the water-fed animals (upper panel of FIG. 4). It is well known that the hippocampus of the brain is a key area involved in learning, memory and cognition. To see if IPA was effective in stimulating PGC-1 alpha expression, the present invention used an in vitro cell culture system, different concentrations of IPA were added to the culture medium of mouse hippocampal neuron cell line HT-22, after 3 days of co-culture the lysed cells were harvested and tested by immunoblotting of PGC-1 alpha, which confirmed that IPA dose-dependently increased PGC-1 alpha protein expression in hippocampal neuron cell line HT-22 cells (FIG. 4, lower panel).

In order to solve the problem that whether the PGC-1 alpha protein in the hippocampal region of the brain can reduce the occurrence of POD (peroxidase), AAV-hDlx-PGC-1 alpha-2 AmCherry recombinant adeno-associated virus is constructed, then the PGC-1 alpha recombinant and AAV-hDlx-2A-mCherry empty vector are respectively fed to a mouse fed with ampicillin, and after the anesthesia operation is carried out and the behavior function score of the mouse is carried out, the result proves that the POD number of the mice in an operation group receiving the anesthesia and operation treatment in the mice receiving the empty vector is obviously increased; however, the amount of POD occurring in the mice given the PGC-1. Alpha. Gene was not significantly different from that in the group to which the AAV vector was injected. PGC-1. Alpha. Was also suggested to have preventive and therapeutic effects on POD (FIG. 5).

In summary, the present inventors have found that the gut microbiota metabolite IPA is inversely related to the development of human POD. In preclinical animal models for POD assessment, IPA deficiency promotes POD-like behavior, while exogenous IPA administration prevents POD-like behavior. This protective effect of IPA is mediated in part by PGC-1 alpha and interneurons in the hippocampus. These results provide a mechanistic view of POD pathogenesis, facilitating the formulation of a principled prophylactic strategy.

It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the present disclosure without departing from the scope or spirit of the disclosure. Other embodiments will be apparent to those skilled in the art from consideration of the specification. The specification and examples are exemplary only.

Claims

Use of pgc-1 α or a production promoter thereof for the preparation of a medicament for the prevention and/or treatment of postoperative delirium.
2. Use according to claim 1, wherein the prevention and/or treatment is cure, alleviation, palliation, prevention of the occurrence or progression of post-operative delirium or post-operative delirium-like behavior.
3. The use according to claim 1, wherein the promoter of PGC-1 α production comprises a tryptophan-related metabolite of the gut microflora.
4. The use according to claim 3, wherein the metabolite comprises indolpropion or an analogue thereof.
Use of pgc-1 a and/or indolpropion in the manufacture of an agent for predicting the risk of post-operative delirium occurrence.
6. Use according to claim 5, characterized in that said prediction comprises:

(1) A step of measuring the content of PGC-1 α and/or indolpropanoic acid in a sample collected from the subject using an agent to obtain a measured value;

(2) A step of comparing the measured value with a standard value;

(3) When the measurement values are below the normative values, then detecting the subject as having, or predicting the subject as being at risk for having, post-operative delirium; when the measurement value is higher than the normative value, then the subject is detected as not suffering from post-operative delirium, or the subject is predicted as not being at risk of suffering from post-operative delirium.
7. The use according to claim 6, wherein the standard value is a value obtained from a biological sample of a normal subject comparable to the age of the subject, or the standard value is a value obtained from a biological sample of the same subject prior to surgery.
8. Use according to claim 6, wherein the amount in the sample T1 taken from the subject at a first point in time is measured as the standard value and the amount in the sample T2 taken from the same subject at a second point in time is measured as the measured value.
9. A method of screening for a compound useful for treating post-operative delirium comprising:

a. a step of measuring the content of PGC-1 α and/or indolpropion in a sample collected pre/post-operatively from a subject suffering from post-operative delirium or post-operative delirium-like behavior to obtain a first measurement value.

b. A step of administering the compound to the subject.

c. A step of measuring the content of PGC-1 α and/or indolpropanoic acid in a sample collected from said subject following administration of said compound to yield a second measurement value.

d. A step of comparing the first measurement value and the second measurement value;

e. selecting the compound as a compound not useful for treating or slowing postoperative delirium or postoperative delirium-like behavior when the second measurement is less than or equal to the first measurement, and selecting the compound as a compound useful for treating or slowing postoperative delirium or postoperative delirium-like behavior when the second measurement is greater than the first measurement.
10. System for predicting the risk of occurrence of post-operative delirium, comprising:

a data acquisition unit for acquiring at least PGC-1 α and/or indole propionic acid content data from a subject; and a judging unit for classifying the subject as belonging to a patient with a high or low risk of developing postoperative delirium based on the result of comparing the content of PGC-1 alpha and/or indolpropion with a preset threshold value.