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Application of senescence markers and caloric restrictions of multiple tissue organs and cell types in delaying senescence of organism

An aging and body technology, applied in the direction of instruments, scientific instruments, recombinant DNA technology, etc., can solve the problem of lack of multi-organ senescent cell atlas

Active Publication Date: 2020-06-02
INST OF ZOOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is still a lack of comprehensive multi-organ senescent cell atlas, and no cross-tissue CR cell atlas.

Method used

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  • Application of senescence markers and caloric restrictions of multiple tissue organs and cell types in delaying senescence of organism
  • Application of senescence markers and caloric restrictions of multiple tissue organs and cell types in delaying senescence of organism
  • Application of senescence markers and caloric restrictions of multiple tissue organs and cell types in delaying senescence of organism

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0096] Example 1. Effects of Aging and CR Diet on Lifespan and Phenotypic Traits

[0097] 1. Experimental groups and methods

[0098] 1. Experimental grouping

[0099] The experiment was divided into three groups, the young group (referred to as the young group, 30 rats, 15 males and 15 females), the old free diet group (abbreviated as the old group, 30 rats, 14 males and 16 females) and the old 70% calorie diet group. In the food-restricted group (referred to as the aged-restricted food group, 26 rats, 12 males and 14 females), each rat was reared in a single cage. The specific treatment methods for each group are as follows:

[0100] Young group: 2-month-old rats were fed with sufficient standardized rat feed every day (to ensure that the feed remained uninterrupted), and histological detection was performed after continuous feeding for 3 months.

[0101] Elderly group: 18-month-old rats were fed with sufficient standardized rat feed every day (to ensure that the feed rem...

Embodiment 2

[0154] Example 2. Effect and verification of aging and CR diet on cell ratio changes

[0155] 1. Experimental method

[0156] In order to describe the compositional changes of dynamic cell types during aging and CR, the following seven tissues were compared between the young group, the old group and the old food-restricted group: brown adipose tissue (BAT), white adipose tissue (WAT), The proportion of each cell type in liver tissue (Liver), kidney tissue (Kidney), aorta (Aorta), skin (Skin) and bone marrow (Bone marrow), and bioinformatics data through immunofluorescence experiments and flow cytometry experiments Prompt cells with changes for experimental verification. Specific steps are as follows:

[0157] 1. Tissue cell isolation and sequencing

[0158] 1) Isolation and sequencing of white fat, brown fat, kidney, skin, liver and vascular tissue cells

[0159] Each group of rats in Example 1 was anesthetized and perfused with physiological saline. After the same tissues...

Embodiment 3

[0184] Example 3. Analysis and verification of key differential genes between aging and CR diet

[0185] 1. Experimental method

[0186] 1. Gene differential expression analysis

[0187] 1) The Wilcoxon rank sum test in the "FindMarkers" function of the Seurat software package was used to perform differential expression analysis of each cell type between the different groups (old / young and old / old restricted food groups). Cell types (B cells in the aorta, CD8 + T cells and plasma cells; B cells, NK cells and neutrophils in the kidney; plasma cells in brown fat; NKT cells in the skin).

[0188] 2) Compare the differentially expressed genes (DEGs) between the aged group and the young group to establish an aging-related DEGs data set (aging DEGs) (|LogFC|>0.5, adjusted P-value0.5, adjustedP-value<0.05). Based on the above 2 datasets, "rescue DEGs", i.e. genes that were significantly upregulated or downregulated in agingDEGs and significantly downregulated or upregulated after ...

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Abstract

The invention discloses an application of senescence markers and caloric restrictions of multiple tissue organs and cell types in delaying senescence of organism. According to the invention, integration analysis of a multi-tissue single-cell transcriptome is carried out by taking a rat as a model so as to research the regulation and control effects of senescence and calorie limitation on a mammalsingle-cell level, and 160000 single cells in seven tissues of a young diet group, an old group and an old diet limitation group are systematically analyzed; based on a high-throughput single-cell transcriptome sequencing technology, a first mammal aging and CR multi-tissue organ high-throughput single-cell transcriptome map is established, the effects of senescence and CR on different tissues andcell types are comprehensively and systematically evaluated in the aspects of cell type composition, cell and tissue specificity differential expression gene, core regulation and control transcription factors, cell-cell communication networks and the like, the complex processes of senescence and CR are revealed, and a basis is provided for systematically researching the molecular regulation and control mechanisms of senescence and CR in the future.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to the application of novel senescence markers of multiple tissues, organs and cell types and calorie restriction in delaying the aging of the body from the levels of multiple tissues, cells and molecules. Background technique [0002] The aging process is accompanied by a decline in the function of multiple tissues in the body and exhibits increased susceptibility to various chronic diseases. At the same time, aging can induce differential responses between different types of cells, tissues and organs, and may even generate heterogeneity in gene expression within the same cell type. On the other hand, in recent decades, more and more researchers have conducted various explorations on how to intervene in aging, among which caloric restriction (CR) is considered to be one of the safest and most effective measures to delay aging . Due to the complexity of the aging process, i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888G01N33/68C12N15/113G01N21/84
CPCC12Q1/6888G01N33/68G01N33/6863C12N15/113G01N21/84C12Q2600/158G01N2333/47G01N2333/525C12N2310/14C12N2310/531
Inventor 刘光慧曲静张维绮马帅孙淑慧耿令令宋默识王思
Owner INST OF ZOOLOGY CHINESE ACAD OF SCI
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