Limits...
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.

Gu H, Tang C, Peng K, Sun H, Yang Y - J. Biomed. Biotechnol. (2009)

Bottom Line: Here, we investigated the effect of chronic mild stress (CMS) on the development of atherosclerosis as well as the expression of Toll-like receptors (TLRs) signaling pathway in adolescent apolipoprotein E knockout (apoE-/-) mice.We found that atherosclerosis lesions both in aortic tress and sinuses of CMS mice were significantly increased linearly in response to duration of CMS exposure.CMS mice demonstrated markedly increased aortic atherosclerosis that were associated with significant increases in levels of expression of TLR4, MyD88, nuclear factor kappaB (NF-kappaB), MCP-1, IL-1beta, TNF-alpha, and sICAM-1.

View Article: PubMed Central - PubMed

Affiliation: Key Lab for Arteriosclerology of Hunan Province, Institute of Cardiovascular Disease, University of South China, Hengyang, China.

ABSTRACT
Here, we investigated the effect of chronic mild stress (CMS) on the development of atherosclerosis as well as the expression of Toll-like receptors (TLRs) signaling pathway in adolescent apolipoprotein E knockout (apoE-/-) mice. Mice were subjected to daily CMS for 0, 4, and 12 weeks, respectively. To identify the expression of Toll-like receptor 4 signaling pathway in adolescent apolipoprotein E knockout mice subjected to CMS, we compared gene expression in aortas of stressed and unstressed mice using TLRs signaling pathway real-time PCR microarrays consisting of 87 genes. We found that atherosclerosis lesions both in aortic tress and sinuses of CMS mice were significantly increased linearly in response to duration of CMS exposure. Among 87 genes analyzed, 15 genes were upregulated in stressed mice, especially TLR4, myeloid differentiation factor 88 (MyD88), and IL-1beta, and 28 genes were downregulated compared with nonstressed mice. CMS mice demonstrated markedly increased aortic atherosclerosis that were associated with significant increases in levels of expression of TLR4, MyD88, nuclear factor kappaB (NF-kappaB), MCP-1, IL-1beta, TNF-alpha, and sICAM-1. Taken together, our results suggest an important role for TLR4 signaling pathway in atherosclerosis in a CMS mouse model.

Show MeSH

Related in: MedlinePlus

Validation of changes in TLR-4 (73 KD) and NF-κB p65 (80 kDa) protein expression by western blotting. Proteins extracted were prepared from arteries (n = 6) of mice subjected to CMS or normal conditions for 0, 4, and 12 weeks, respectively. Each lane shows representative western blots using anti-TLR4 or NF-κB p65 and anti-GAPDH bodies in (a1) and (a2), respectively. Each panel summarizes densitometric readings of band intensities normalized to GAPDH, which was measured by densitometry with Image J image analysis software.  (b1), (b2) densitometric measurements TLR4 and NF-κB p65 from Western blots, respectively. Data are mean  ± SEM. C: control group; S: chronic mild stress group (n = 6 per group). *P < .05, **P < .01 compared with the control. The data are representative of three experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2734999&req=5

fig6: Validation of changes in TLR-4 (73 KD) and NF-κB p65 (80 kDa) protein expression by western blotting. Proteins extracted were prepared from arteries (n = 6) of mice subjected to CMS or normal conditions for 0, 4, and 12 weeks, respectively. Each lane shows representative western blots using anti-TLR4 or NF-κB p65 and anti-GAPDH bodies in (a1) and (a2), respectively. Each panel summarizes densitometric readings of band intensities normalized to GAPDH, which was measured by densitometry with Image J image analysis software. (b1), (b2) densitometric measurements TLR4 and NF-κB p65 from Western blots, respectively. Data are mean ± SEM. C: control group; S: chronic mild stress group (n = 6 per group). *P < .05, **P < .01 compared with the control. The data are representative of three experiments.

Mentions: Western blotting with an Ab against TLR4 and NF-κB showed reactive protein bands at the size of TLR4 (73 kDa) and NF-κB p65 (80 kDa) in aortas of chronic mild stress and control groups following different weeks of CMS. As demonstrated in Figure 6, TLR4 and NF-κB protein levels in mice subjected to 4 and 12 weeks chronic mild stress were more abundant compared with that in the corresponding control group mice, consistent with the mRNA pattern (Table 2(a)). Thus, both protein and mRNA of TLR4 and NF-κB p65 were more abundant in CMS mice than that in control mice, respectively (P < .05, P < .01). In Figure 6(b(1)) a mean relative intensity of TLR4 in CMS mice was stronger than that in control mice (P < .05, P < .01).


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.

Gu H, Tang C, Peng K, Sun H, Yang Y - J. Biomed. Biotechnol. (2009)

Validation of changes in TLR-4 (73 KD) and NF-κB p65 (80 kDa) protein expression by western blotting. Proteins extracted were prepared from arteries (n = 6) of mice subjected to CMS or normal conditions for 0, 4, and 12 weeks, respectively. Each lane shows representative western blots using anti-TLR4 or NF-κB p65 and anti-GAPDH bodies in (a1) and (a2), respectively. Each panel summarizes densitometric readings of band intensities normalized to GAPDH, which was measured by densitometry with Image J image analysis software.  (b1), (b2) densitometric measurements TLR4 and NF-κB p65 from Western blots, respectively. Data are mean  ± SEM. C: control group; S: chronic mild stress group (n = 6 per group). *P < .05, **P < .01 compared with the control. The data are representative of three experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2734999&req=5

fig6: Validation of changes in TLR-4 (73 KD) and NF-κB p65 (80 kDa) protein expression by western blotting. Proteins extracted were prepared from arteries (n = 6) of mice subjected to CMS or normal conditions for 0, 4, and 12 weeks, respectively. Each lane shows representative western blots using anti-TLR4 or NF-κB p65 and anti-GAPDH bodies in (a1) and (a2), respectively. Each panel summarizes densitometric readings of band intensities normalized to GAPDH, which was measured by densitometry with Image J image analysis software. (b1), (b2) densitometric measurements TLR4 and NF-κB p65 from Western blots, respectively. Data are mean ± SEM. C: control group; S: chronic mild stress group (n = 6 per group). *P < .05, **P < .01 compared with the control. The data are representative of three experiments.
Mentions: Western blotting with an Ab against TLR4 and NF-κB showed reactive protein bands at the size of TLR4 (73 kDa) and NF-κB p65 (80 kDa) in aortas of chronic mild stress and control groups following different weeks of CMS. As demonstrated in Figure 6, TLR4 and NF-κB protein levels in mice subjected to 4 and 12 weeks chronic mild stress were more abundant compared with that in the corresponding control group mice, consistent with the mRNA pattern (Table 2(a)). Thus, both protein and mRNA of TLR4 and NF-κB p65 were more abundant in CMS mice than that in control mice, respectively (P < .05, P < .01). In Figure 6(b(1)) a mean relative intensity of TLR4 in CMS mice was stronger than that in control mice (P < .05, P < .01).

Bottom Line: Here, we investigated the effect of chronic mild stress (CMS) on the development of atherosclerosis as well as the expression of Toll-like receptors (TLRs) signaling pathway in adolescent apolipoprotein E knockout (apoE-/-) mice.We found that atherosclerosis lesions both in aortic tress and sinuses of CMS mice were significantly increased linearly in response to duration of CMS exposure.CMS mice demonstrated markedly increased aortic atherosclerosis that were associated with significant increases in levels of expression of TLR4, MyD88, nuclear factor kappaB (NF-kappaB), MCP-1, IL-1beta, TNF-alpha, and sICAM-1.

View Article: PubMed Central - PubMed

Affiliation: Key Lab for Arteriosclerology of Hunan Province, Institute of Cardiovascular Disease, University of South China, Hengyang, China.

ABSTRACT
Here, we investigated the effect of chronic mild stress (CMS) on the development of atherosclerosis as well as the expression of Toll-like receptors (TLRs) signaling pathway in adolescent apolipoprotein E knockout (apoE-/-) mice. Mice were subjected to daily CMS for 0, 4, and 12 weeks, respectively. To identify the expression of Toll-like receptor 4 signaling pathway in adolescent apolipoprotein E knockout mice subjected to CMS, we compared gene expression in aortas of stressed and unstressed mice using TLRs signaling pathway real-time PCR microarrays consisting of 87 genes. We found that atherosclerosis lesions both in aortic tress and sinuses of CMS mice were significantly increased linearly in response to duration of CMS exposure. Among 87 genes analyzed, 15 genes were upregulated in stressed mice, especially TLR4, myeloid differentiation factor 88 (MyD88), and IL-1beta, and 28 genes were downregulated compared with nonstressed mice. CMS mice demonstrated markedly increased aortic atherosclerosis that were associated with significant increases in levels of expression of TLR4, MyD88, nuclear factor kappaB (NF-kappaB), MCP-1, IL-1beta, TNF-alpha, and sICAM-1. Taken together, our results suggest an important role for TLR4 signaling pathway in atherosclerosis in a CMS mouse model.

Show MeSH
Related in: MedlinePlus