Effects of Chronic Restraint Stress on Apoptosis of Amygdala Cells in Rats

doi:10.12116/j.issn.1004-5619.2021.410101

Abstract

Abstract:

Objective To explore the damage effects of chronic restraint stress （CRS） on amygdala cells through the rat CRS model. Methods The rat CRS model was established, and the changes in body weight and adrenal mass in control group and CRS group were monitored at 1 d, 7 d, 14 d and 21 d. The behavior changes were evaluated by the percentage of retention time of open arms and open arm entries using the elevated plus maze （EPM）. ELISA was used to detect the concentrations of rat’s corticotropin releasing hormone （CRH）, adrenocorticotropic hormone （ACTH） and cortisol. The changes of expression of glucocorticoid receptor （GR） and glial fibrillary acidic protein （GFAP） in amygdala were determined by immunohistochemistry and Western blotting. Ultrastructure changes of glial cell were observed by transmission electron microscopy. The apoptosis rate of amygdala was measured by flow cytometry. Results Compared with the control group at the same time points, body weight of CRS 1 d, 7 d, 14 d and 21 d groups increased slowly, but adrenal mass increased significantly; the serum level of CRH, cortisol and ACTH increased significantly at 7 d, 14 d and 21 d respectively; the expression of GR in amygdala was increased while that of GFAP was decreased; EPM test suggested that the percentage of retention time of open arms and open arm entries decreased significantly after 14 d. The CRS group showed different degrees of glial cell damage in amygdala, and the apoptosis rate of glial cell was significantly increased in 21 d group. Conclusion This study successfully established a CRS model in rats, and anxiety-like behavioral changes in model rats may be caused by apoptosis of amygdala astrocytes.

Key words: forensic pathology, animal model, amygdala, apoptosis, elevated plus maze, enzyme linked immunosorbent assay, Western blotting, chronic restraint stress, rats

CLC Number:

DF795.1

Li-ru DONG, Jun-bo LIAN, Shuang-jie HUO, Dan LUO, Hu YANG, Xu-dong SONG, Xiao-jing ZHANG, Bin CONG. Effects of Chronic Restraint Stress on Apoptosis of Amygdala Cells in Rats[J]. Journal of Forensic Medicine, 2022, 38(4): 459-467.

Figures/Tables 9

References 28

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时间点	对照组	CRS组
0 d	258.00±7.48	262.38±11.89
1 d	273.63±12.13	261.88±6.17^1）
7 d	304.75±12.43	277.00±16.64^1）
14 d	330.50±15.64	303.25±18.99^1）
21 d	361.38±19.32	309.62±19.08^1）

时间点	对照组	CRS组
0 d	258.00±7.48	262.38±11.89
1 d	273.63±12.13	261.88±6.17^1）
7 d	304.75±12.43	277.00±16.64^1）
14 d	330.50±15.64	303.25±18.99^1）
21 d	361.38±19.32	309.62±19.08^1）

组别	总进臂次数	总运动距离/mm	开臂停留时间百分比/%	进入开臂次数百分比/%
对照	14.72±8.84	22 852.91±7 578.87	10.94±5.64	27.98±13.09
CRS 1 d	13.80±8.58	20 362.41±2 630.74	9.82±3.54	16.71±4.76
CRS 7 d	15.50±3.78	23 585.06±3 474.36	5.10±3.30^1）	10.82±2.39^1）
CRS 14 d	16.50±11.20	23 095.60±2 808.39	4.62±4.66^1）	12.23±3.09^1）
CRS 21 d	12.70±6.60	19 038.74±5 628.17	3.00±1.12^1）	12.22±6.45^1）

组别	总进臂次数	总运动距离/mm	开臂停留时间百分比/%	进入开臂次数百分比/%
对照	14.72±8.84	22 852.91±7 578.87	10.94±5.64	27.98±13.09
CRS 1 d	13.80±8.58	20 362.41±2 630.74	9.82±3.54	16.71±4.76
CRS 7 d	15.50±3.78	23 585.06±3 474.36	5.10±3.30^1）	10.82±2.39^1）
CRS 14 d	16.50±11.20	23 095.60±2 808.39	4.62±4.66^1）	12.23±3.09^1）
CRS 21 d	12.70±6.60	19 038.74±5 628.17	3.00±1.12^1）	12.22±6.45^1）

组别	CRH	ACTH	皮质醇
对照	10.57±1.74	11.52±0.72	2.87±1.79
CRS 1 d	9.83±0.97	12.55±1.26	3.15±1.82
CRS 7 d	13.67±0.53^1）	11.19±1.38	3.38±3.01
CRS 14 d	13.61±0.57^1）	12.22±0.81	6.03±3.00^1）
CRS 21 d	13.87±1.44^1）	14.61±0.71^1）	8.40±7.42^1）