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Quad-synapsis complex cell model of diabetic concurrent depression and building method and application thereof

A technology of cell model and establishment method, applied in the field of cell model, which can solve the problems of insufficient simulation of in vivo environment and difficulty of real response

Inactive Publication Date: 2019-06-21
HUNAN UNIV OF CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The cell model of this application only examines the response of hippocampal neurons in diabetes complicated with depression, and the simulation degree of the in vivo environment is not enough, so it is difficult to truly reflect the changes in the body

Method used

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  • Quad-synapsis complex cell model of diabetic concurrent depression and building method and application thereof
  • Quad-synapsis complex cell model of diabetic concurrent depression and building method and application thereof
  • Quad-synapsis complex cell model of diabetic concurrent depression and building method and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] The cell models after co-cultivation for 3 days were randomly divided into normal control group, high glucose group, corticosterone group, and model group. Multiple wells were set up in each group, and corresponding intervention drugs were added according to the above grouping details, among which the high glucose group was added Glucose at a final concentration of 150 mM, corticosterone at a final concentration of 200 μM was added to the corticosterone group, and glucose at a final concentration of 150 mM and corticosterone at a final concentration of 200 μM were added to the high-glucose combined corticosterone group for 18 hours. The general morphology of hippocampal neurons, astrocytes and microglia under intervention conditions were observed separately.

[0045] The experimental results showed that in the normal control group, the NE cell body was round and bright, with good gloss and refraction, with criss-cross protrusions and numerous small branches, which were i...

Embodiment 2

[0047] Enzyme-linked immunosorbent assay (Elisa) was used to detect the contents of neurotransmitters NE, DA, and 5-HT in hippocampal neurons to evaluate whether the tested cells produced depression-like indicator characteristics. The specific operation is as follows: After co-cultivating the cell model for 3 days, the cells were divided into normal control group, high glucose group, corticosterone group, high glucose combined with corticosterone group, and each group was set with 4 replicate wells. The high-glucose group added glucose with a final concentration of 150 mM, the corticosterone group added corticosterone with a final concentration of 200 μM, and the high-glucose combined corticosterone group added glucose with a final concentration of 150 mM and corticosterone with a final concentration of 200 μM. Wash 3 times with cold PBS, add RIPA cell lysate containing PMSF, inoculate on a sterile ice box at 4°C for 1 hour, centrifuge at 12000 for 15 minutes, collect the super...

Embodiment 3

[0053] Inoculate hippocampal neurons into a 96-well plate, and inoculate mixed glial cells into another 96-well plate at the same time, wait for the cells to grow to 5-7 days, and when the neural network and glial cells grow to about 80% confluence respectively , except that the blank control group was given the same amount of maintenance medium, the corresponding concentrations of high glucose (150mM) and corticosterone (200μM) were added to the glial cells of the other groups, and three replicate wells were set up in each group. After 18 hours of modeling intervention, , take the supernatant to neurons, and then incubate in a 37°C incubator for 18h. Add 20 μL MTT to each well, act for 4 hours, discard the liquid, add 150 μL DMSO to each well, shake on a horizontal shaker for 15 minutes, and finally measure the OD value of each well with a microplate reader at a wavelength of 492 nm, and calculate the relative survival rate of cells in each group .

[0054] The results showe...

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Abstract

The invention belongs to a cell model, and particularly relates to a quad-synapsis complex cell model of diabetic concurrent depression and a building method and application thereof. The building method includes: adding glucose and corticosterone into a quad-synapsis co-culture system, and intervening to acquire the quad-synapsis complex cell model, wherein the quad-synapsis co-culture system comprises mixed glial cells and hippocampal neuron positioned on the mixed glial cells, and the mixed glial cells comprise mixed cells of starlike glial cells and microglial cells. The quad-synapsis complex cell model is used for pathophysiological study, pathogenesis study and in-vitro drug screening of diseases related to diabetic concurrent depression.

Description

technical field [0001] The invention belongs to a cell model, and in particular relates to a quadruple synaptic compound cell model of diabetes complicated with depression, a method for establishing it and an application thereof. Background technique [0002] Diabetes Mellitus (DM) is a chronic metabolic disease characterized by hyperglycemia. With the aging of the population in my country, its incidence is increasing day by day. According to the statistics of the International Diabetes Federation in 2017, there are about 425 million diabetic patients in the world, and 114.4 million in my country. head. Diabetes Mellitus with Depression (DD), with cognitive impairment as the main clinical manifestation, is one of the common chronic complications of DM. According to the latest research results, DM patients with depression are as high as 60-75%, of which severe depression accounts for 10%-35%. The mortality rate of DD patients is significantly higher than that of simple DM pa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0793C12Q1/02
Inventor 王宇红刘检凌佳韩远山
Owner HUNAN UNIV OF CHINESE MEDICINE
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