CN-121068734-B - ELS-induced depression-like behavior rat whole brain tissue lipid space metabolism analysis method

CN121068734BCN 121068734 BCN121068734 BCN 121068734BCN-121068734-B

Abstract

The application relates to the technical field of biomedicine, in particular to a lipid space metabolism analysis method for whole brain tissue of an ELS-induced depression-like behavior rat, which comprises the following steps of firstly constructing an ELS-induced depression-like behavior rat model, determining a depression model group by adopting a depression-like behavioral test, then taking a conventional feeding rat as a control group, respectively carrying out 2D scanning on the control group and the depression model group based on a mass spectrum space histology technology, constructing a 2D brain lipid map, carrying out whole brain 3D reconstruction to obtain brain tissue lipid mass spectrum imaging data, and finally carrying out metabolic heterogeneity analysis on the brain tissue lipid mass spectrum imaging data. The method provided by the application aims to provide technical and data map support for the research of the molecular mechanism of lipid metabolic disorder of early negative stress induced depression occurrence and development from the animal model level, and simultaneously expands the application range of mass spectrum space histology technology.

Inventors

ZHANG ZHIJUN
LUO QIAN
ZHAO CHAO
GAO CHENYU
YANG ZHIYI
CAO TIANYOU

Assignees

深圳先进技术研究院

Dates

Publication Date: 20260508
Application Date: 20240604

Claims (6)

1. A method for analyzing lipid space metabolism of whole brain tissue of an ELS-induced depression-like behavior rat, comprising the steps of: Adopting a maternal-generation gestation period constraint stress combined offspring maternal-infant separation and growth development period chronic unpredictable mild stimulation method to construct an ELS-induced depression-like behavior rat model, and adopting a depression-like behavioral test to determine a depression model group; preparing 14 mu m brain tissue slices of a control group and a depression model group respectively, and preprocessing the brain tissue slices, wherein the brain tissue slices are continuous frozen slices of brain tissue coronal planes; 2D scanning is carried out on brain tissue slices of the control group and the depression model group by using a conventional feeding rat as a control group and based on a mass spectrometry space histology technology by adopting MALDI-TOF MSI equipment, continuous brain slice data of the control group and the depression model group are imported into SCiLS Lab software to carry out full brain 3D reconstruction, the z coordinate axis position is set, namely the distance between adjacent slices is set to be 2mm in the reconstruction process, then a center point is set in the brain tissue slices positioned on a glass slide, continuous alignment is achieved by multi-angle rotation adjustment based on the center point, and full brain 3D reconstruction is carried out to obtain brain tissue lipid mass spectrometry imaging data; And (3) carrying out metabolic heterogeneity analysis on the brain tissue lipid mass spectrometry imaging data to obtain characteristic lipid molecules PA (42:8) and characteristic lipid molecules PE (40:2) which show significant spatial distribution differences in a plurality of functional areas, and characterizing the spatial distribution heterogeneity of the functional brain areas of the ELS-induced depression-like behavior rat from the lipid metabolism heterogeneity level, wherein the characteristic lipid molecules PA (42:8) and the characteristic lipid molecules PE (40:2) have specific enrichment in the prefrontal cortex and have specific deletion in the nucleus accumbens.
2. The ELS-induced depression-like behavioural rat brain tissue lipid space metabolic analysis method of claim 1, wherein the depression-like behavioural test comprises a sugar water preference test, a open field test and a forced swimming test.
3. The ELS-induced depression-like behavioural rat whole brain tissue lipid spatial metabolic analysis method according to claim 1, characterized in that determining a depression model group using a depression-like behavioural test comprises: and determining a depression model group by evaluating depression-like behaviors, wherein if rats show reduced sugar water preference rate and prolonged forced swimming immobility time, the modeling is successful, and the model group with depression-like behaviors is used as the depression model group.
4. The ELS-induced depression-like behavior rat whole brain tissue lipid spatial metabolic analysis method of claim 1, wherein metabolic heterogeneity analysis is performed on the brain tissue lipid mass spectrometry imaging data, comprising: carrying out image data segmentation processing on the brain tissue lipid mass spectrometry imaging data; And analyzing lipid metabolites of which the expression is up-regulated and down-regulated by rats in the depression model group based on the image data after the segmentation treatment by using the control group as a comparison.
5. The ELS-induced depression-like behavior rat whole brain tissue lipid spatial metabolic analysis method according to claim 4, wherein the processing of the segmentation of the brain tissue lipid mass spectrometry imaging data into picture data comprises: Sequentially carrying out mass spectrum peak searching, peak alignment and noise reduction treatment on the brain tissue lipid mass spectrum imaging data to obtain treated picture data; And carrying out image segmentation processing on the processed image data by adopting K-Means.
6. The method for analyzing lipid space metabolism of whole brain tissue of ELS-induced depression-like behavior rat according to claim 4, wherein analyzing lipid metabolites up-regulated and down-regulated in rats in the depression model group based on the image data after the segmentation treatment by using the control group as a comparison, comprises: Carrying out multi-element statistical analysis on the space data of the segmented picture data, and screening characteristic lipid molecules under each principal component condition to obtain the space distribution difference of characteristic lipid molecules PA and PE in a plurality of functional areas; The spatial distribution heterogeneity of the ELS-induced depression-like behavioural rat functional brain region is characterized from the lipid metabolism heterogeneity level based on the spatial distribution differences exhibited by the characteristic lipid molecules PA and the characteristic lipid molecules PE in the multiple functional regions.

Description

ELS-induced depression-like behavior rat whole brain tissue lipid space metabolism analysis method Technical Field The application relates to the technical field of biomedicine, in particular to a lipid space metabolism analysis method for rat brain tissue with depression-like behavior induced by early life stress (EARLY LIFE STRESS, ELS). Background Depressive disorder is the most common heavy mental disease, the total burden of the disease is high and the disease is the second most in human, the disease has a trend of low age in recent years, according to the latest national mental health development report (2021-2022), 14.8% of teenagers in China currently have different degrees of depressive states, wherein the major depression is as high as 4.0%, and the depression risk has a trend of rising along with the increase of the grade. It is well known that early life is a critical period of brain neural development plasticity, and adverse experiences such as stress exposure are highly sensitive, and the risk of occurrence of depressive disorder is sharply increased, however, the exact pathogenesis thereof has not yet been elucidated. The research shows that lipid metabolism of brain tissues of patients with depression disorder is abnormal, and can cause neurotransmitter dysfunction and neuroimmune inflammation in brain, whether key lipid metabolism pathway molecules are starting factors of depression occurrence needs to be clarified, and the adoption of a spatial lipid metabonomics comprehensive system for screening key lipid metabolism molecules of rats anchoring depression-like behavior patterns is significant for clarifying pathogenesis and clinical application. Currently, research on depressive disorder brain lipid metabolism mainly depends on a homogenate system analysis method, such as a liquid chromatography-mass spectrometry (LC-MS/MS) and gas chromatography-mass spectrometry based technology. The prior art mainly focuses on two aspects, namely 1, analyzing the lipid metabolism of brain tissues of a homogenate system by adopting a mass spectrometry technology, realizing qualitative and quantitative measurement of lipid molecules, but losing in-situ space positioning information of the lipid, and 2, researching lipid metabolites of brain slices by adopting a mass spectrometry space histology technology, and realizing qualitative, quantitative and space positioning of the lipid molecules by adopting a 2D level mass spectrometry imaging. However, there are numerous brain regions, biological functional differences exist between the brain regions, and conventional 2D level mass spectrometry imaging cannot satisfy in-situ spatial distribution research on lipid metabolites in multiple functional brain regions simultaneously. Disclosure of Invention The embodiment of the application provides a lipid space metabolism analysis method for rat brain tissue with ELS-induced depression-like behaviors, which aims at developing the study on the spatial distribution of lipid molecules in different brain regions by taking the brain tissue of a rat model with ELS-induced depression-like behaviors as a study object by utilizing 2D and 3D mass spectrum space histology technologies, and widens the application range of the mass spectrum space histology technologies and the tissue morphology technologies in the clinical medicine and accurate diagnosis fields. In order to solve the technical problems, the embodiment of the application provides a lipid space metabolism analysis method for whole brain tissues of an ELS-induced depression-like behavior rat, which comprises the following steps of firstly constructing an ELS-induced depression-like behavior rat model, determining a depression model group by adopting a depression-like behavioral test, then carrying out 2D scanning on the control group and the depression model group by taking a conventional feeding rat as a control group and based on a mass spectrum space histology technology, constructing a 2D brain lipid map, carrying out whole brain 3D reconstruction to obtain brain tissue lipid mass spectrum imaging data, and finally carrying out metabolic heterogeneity analysis on the brain tissue lipid mass spectrum imaging data. In some exemplary embodiments, an ELS-induced depression-like behavioral rat model is constructed using maternal-to-maternal gestation period constraint stress in combination with maternal-to-infant segregation and chronic unpredictable warmth and stimulation methods of growth and development. In some exemplary embodiments, the depression-like behavioral tests include a sugar water preference test, a open field test, and a forced swim test. In some exemplary embodiments, determining a depression model set using a depression-like behavioral test includes determining a depression model set by evaluating depression-like behavior, if the rat exhibits a reduced sugar water preference rate and prolonged forced swimming immobility time, indicating successfu