Effects of Chronic Mild Stress on the Development of Atherosclerosis and Expression of Toll-Like Receptor 4 Signaling Pathway in Adolescent Apolipoprotein E Knockout Mice

Table 2

CMS alters TLRs signaling pathway gene expression profile in aortas of apoE-/- mice. ApoE-/- mice 4 weeks old were subjected to a 24-hour CMS daily for 0, 4, 12 weeks. Mice were sacrificed, and aortas were harvested. Total RNA was extracted, and microarray analysis was determined as described under “materials and methods.” TLRs signaling pathway array consisting of 87 were used to analyze gene expression in aorta from mice with or without chronic mild stress. The microarrays were used according to the manufacturer's instructions. The table showed gene expression changes ≥2.0-fold versus control. Calculate the fold change for each gene from control group to stress group as 2-ΔΔCt. Among 87 genes analyzed, 15 genes were upregulated, and 28 genes were downregulated by CMS.

(a) Genes upregulated in aorta of mice exposure to CMS compared with control mice (n  = 5)

Well	Symbol	Description	Fold increase	𝑃-value
A03	Ccl2	Chemokine (C-C motif) ligand 2	5.3	.02
A04	Cd14	CD14 antigen	2.4	.013
A09	Clec4e	C-type lectin domain family 4, member e	3.88	.026
A10	Csf2	Colony stimulating factor 2 (granulocyte-macrophage)	2.06	.018
C03	Il1b	Interleukin 1 beta	11.67	.008
C06	Il6	Interleukin 6	3.54	.018
C08	Irak1	Interleukin-1 receptor-associated kinase 1	2.78	.016
D12	Myd88	Myeloid differentiation primary response gene 88	3.62	.032
E04	Nfkbib	Nuclear factor of kappa light chain gene enhancer in B-cells inhibitor, beta	2.61	.041
E05	Nfkbil1	Nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor-like 1	3.35	.035
E09	Pglyrp1	Peptidoglycan recognition protein 1	5.84	.011
F02	Rela	V-rel reticuloendotheliosis viral oncogene homolog A (avian)	2.17	.031
F11	Tlr4	Toll-like receptor 4	4.04	.014
G05	Tnf	Tumor necrosis factor	2.15	.018
H03	Hsp90ab1	Heat shock protein 90kDa alpha (cytosolic), class B member 1	2.05	.012

(b) Genes downregulated in aorta of mice exposure to CMS compared with control mice (n  = 5)

Well	Symbol	Description	Fold decrease	𝑃-value
A01	Btk	Bruton agammaglobulinemia tyrosine kinase	−3.13	.011
A02	Casp8	Caspase 8	−2.26	.024
A11	Csf3	Colony stimulating factor 3 (granulocyte)	−8.03	.032
B01	Elk1	ELK1, member of ETS oncogene family	−2.47	.021
B03	Fos	FBJ osteosarcoma oncogene	−2.06	.013
C01	Il12a	Interleukin 12A	−2.67	.028
C07	Il6ra	Interleukin 6 receptor, alpha	−2.9	.022
C09	Irak2	Interleukin-1 receptor-associated kinase 2	−2.08	.037
C11	Irf3	Interferon regulatory factor 3	−4.33	.019
C12	Jun	Jun oncogene	−2.06	.042
D01	Lta	Lymphotoxin A	−13.13	.011
D02	Muc13	Mucin 13, epithelial transmembrane	−2.12	.016
D10	Mapk8ip3	Mitogen-activated protein kinase 8 interacting protein 3	−6.89	.027
E02	Nfkb2	Nuclear factor of kappa light polypeptide gene enhancer in B-cells	−2.06	.036
E06	Nfrkb	Nuclear factor related to kappa B binding protein	−4.94	.012
E07	Nr2C2	Nuclear receptor subfamily 2, group C, member 2	−3.24	.028
E08	Peli1	Pellino 1	−2.45	.023
F01	Mapk8	Mitogen-activated protein kinase 8	−2.34	.026
F05	Ticam1	Toll-like receptor adaptor molecule 1	−24.17	.003
F06	Ticam2	Toll-like receptor adaptor molecule 2	−2.4	.044
F08	Tlr1	Toll-like receptor 2	−2.05	.032
F12	Tlr5	Toll-like receptor 5	−6.57	.026
G04	Tlr9	Toll-like receptor 9	−5.37	.022
G06	Tnfaip3	Tumor necrosis factor, alpha-induced protein 3	−4.81	.023
G10	Traf6	Tnf receptor-associated factor 6	−2.52	.034
G11	Ube2n	Ubiquitin-conjugating enzyme E2N	−2.39	.028
H04	Gapdh	Glyceraldehyde-3-phosphate dehydrogenase	−3.26	.032
H05	Actb	Actin, beta, cytoplasmic	−2.63	.025