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C-reactive protein/oxidised low-density lipoprotein/β2-glycoprotein I complex promotes atherosclerosis in diabetic BALB/c mice via p38mitogen-activated protein kinase signal pathway.

Zhang R, Zhou SJ, Li CJ, Wang XN, Tang YZ, Chen R, Lv L, Zhao Q, Xing QL, Yu DM, Yu P - Lipids Health Dis (2013)

Bottom Line: The mRNA expressions of receptors associated with lipid metabolism were quantified by real-time PCR.CRP/oxLDL/β2GPI complex dramatically promoted infiltration of SMCs, macrophages, and T cells, improved the mRNA expression of ABCA1 and ABCG1, but reduced the mRNA expression of SR-BI and CD36 and increased the phosphorylation of p38MAPK and MKK3/6 (all P < 0.05).The highest expression levels of IL-1, IL-9, PF-4, bFGF, and IGF-II were detected in the CRP/oxLDL/β2GPI group (P < 0.05).

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Diabetic Nephropathy Hemodialysis, Key Laboratory of Hormones and Development, Ministry of Health, Metabolic Diseases Hospital & Tianjin Institute of Endocrinology Tianjin Medical University, Tongan Street, Tianjin, Heping District 300070, China.

ABSTRACT

Background: The aim of this study was to investigate the effect of C-reactive protein/oxidised low-density lipoprotein/β2-glycoprotein I (CRP/oxLDL/β2GPI) complex on atherosclerosis (AS) in diabetic BALB/c mice.

Methods: BALB/c mice were fed high-fat and normal diet. Eight weeks later, the mice fed with high-fat diet were injected with streptozotocin (STZ) to induce diabetes. The diabetic mice were respectively injected twice monthly with 20 μg oxLDL, 20 μg β2GPI, 40 μg oxLDL/β2GPI complex, 44 μg CRP/oxLDL/β2GPI complex, and PBS. Aortas were stained with Sudan IV to investigate lipid plaque formation. The infiltration condition of smooth muscle cells (SMCs), macrophages, and T cells in the aortas were determined by immunohistochemistry (IH). The mRNA expressions of receptors associated with lipid metabolism were quantified by real-time PCR. The phosphorylation of p38 mitogen-activated protein kinase (p38MAPK) and MKK3/6 in aorta tissues were assessed by Western blot. The expression of inflammation cytokines was evaluated by protein chip.

Results: The lipid plaques were more extensive, the lumen area was obviously narrower, the ratio of intima and media thickness were increased, and the normal internal elastic lamia structure and endothelial cell disappeared (P < 0.05) in the oxLDL and CRP/oxLDL/β2GPI groups (P < 0.05). CRP/oxLDL/β2GPI complex dramatically promoted infiltration of SMCs, macrophages, and T cells, improved the mRNA expression of ABCA1 and ABCG1, but reduced the mRNA expression of SR-BI and CD36 and increased the phosphorylation of p38MAPK and MKK3/6 (all P < 0.05). The highest expression levels of IL-1, IL-9, PF-4, bFGF, and IGF-II were detected in the CRP/oxLDL/β2GPI group (P < 0.05).

Conclusions: CRP/oxLDL/β2GPI complex aggravated AS in diabetic BALB/c mice by increasing lipid uptake, the mechanism of which may be mediated by the p38MAPK signal pathway.

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Effects of CRP/oxLDL/β2GPI on inflammatory cytokines in the aortas. Extracts of protein from the NC and diabetes animals were added into each well of Protein chip. A laser scanner (Scan-array Gx, PerkinElmer) was used to read the signals for each spot. The relative fold differences in the cytokine amount were determined by an analysis tool (Raybio, USA). A: Protein chip images. Red: positive control; black: bFGF; yellow: IGF-II; pink: IL-1; blue: IL-9; green: PF-4; white: negative control. B: IL-1, IL-9, bFGF, IGF-II and PF-4 average relative optical density in the aortas.
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Figure 7: Effects of CRP/oxLDL/β2GPI on inflammatory cytokines in the aortas. Extracts of protein from the NC and diabetes animals were added into each well of Protein chip. A laser scanner (Scan-array Gx, PerkinElmer) was used to read the signals for each spot. The relative fold differences in the cytokine amount were determined by an analysis tool (Raybio, USA). A: Protein chip images. Red: positive control; black: bFGF; yellow: IGF-II; pink: IL-1; blue: IL-9; green: PF-4; white: negative control. B: IL-1, IL-9, bFGF, IGF-II and PF-4 average relative optical density in the aortas.

Mentions: The mice protein antibody chip detected 24 kinds of cytokines. Each cytokine was represented by duplicate spots, as seen in Table 1. According to the chemiluminescence results, five cytokines were detected (Figure 7A) as follows: basic fibroblast growth factor (bFGF), insulin-like growth factor α (IGF-α), IL-1α, IL-9, and platelet factor-4 (PF-4). Among them, PF-4 was the most predominant. Values were obtained using LabWorks software. These five cytokines were induced to higher levels in the oxLDL/β2GPI group, ranking second to the CRP/oxLDL/β2GPI group (Table 2 and Figure 7B). Average light intensity was normalised without background for each pair of cytokine spots detected using Image Pro-Plus software. Results were represented as bars (n = 3 mice per group).


C-reactive protein/oxidised low-density lipoprotein/β2-glycoprotein I complex promotes atherosclerosis in diabetic BALB/c mice via p38mitogen-activated protein kinase signal pathway.

Zhang R, Zhou SJ, Li CJ, Wang XN, Tang YZ, Chen R, Lv L, Zhao Q, Xing QL, Yu DM, Yu P - Lipids Health Dis (2013)

Effects of CRP/oxLDL/β2GPI on inflammatory cytokines in the aortas. Extracts of protein from the NC and diabetes animals were added into each well of Protein chip. A laser scanner (Scan-array Gx, PerkinElmer) was used to read the signals for each spot. The relative fold differences in the cytokine amount were determined by an analysis tool (Raybio, USA). A: Protein chip images. Red: positive control; black: bFGF; yellow: IGF-II; pink: IL-1; blue: IL-9; green: PF-4; white: negative control. B: IL-1, IL-9, bFGF, IGF-II and PF-4 average relative optical density in the aortas.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3643870&req=5

Figure 7: Effects of CRP/oxLDL/β2GPI on inflammatory cytokines in the aortas. Extracts of protein from the NC and diabetes animals were added into each well of Protein chip. A laser scanner (Scan-array Gx, PerkinElmer) was used to read the signals for each spot. The relative fold differences in the cytokine amount were determined by an analysis tool (Raybio, USA). A: Protein chip images. Red: positive control; black: bFGF; yellow: IGF-II; pink: IL-1; blue: IL-9; green: PF-4; white: negative control. B: IL-1, IL-9, bFGF, IGF-II and PF-4 average relative optical density in the aortas.
Mentions: The mice protein antibody chip detected 24 kinds of cytokines. Each cytokine was represented by duplicate spots, as seen in Table 1. According to the chemiluminescence results, five cytokines were detected (Figure 7A) as follows: basic fibroblast growth factor (bFGF), insulin-like growth factor α (IGF-α), IL-1α, IL-9, and platelet factor-4 (PF-4). Among them, PF-4 was the most predominant. Values were obtained using LabWorks software. These five cytokines were induced to higher levels in the oxLDL/β2GPI group, ranking second to the CRP/oxLDL/β2GPI group (Table 2 and Figure 7B). Average light intensity was normalised without background for each pair of cytokine spots detected using Image Pro-Plus software. Results were represented as bars (n = 3 mice per group).

Bottom Line: The mRNA expressions of receptors associated with lipid metabolism were quantified by real-time PCR.CRP/oxLDL/β2GPI complex dramatically promoted infiltration of SMCs, macrophages, and T cells, improved the mRNA expression of ABCA1 and ABCG1, but reduced the mRNA expression of SR-BI and CD36 and increased the phosphorylation of p38MAPK and MKK3/6 (all P < 0.05).The highest expression levels of IL-1, IL-9, PF-4, bFGF, and IGF-II were detected in the CRP/oxLDL/β2GPI group (P < 0.05).

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Diabetic Nephropathy Hemodialysis, Key Laboratory of Hormones and Development, Ministry of Health, Metabolic Diseases Hospital & Tianjin Institute of Endocrinology Tianjin Medical University, Tongan Street, Tianjin, Heping District 300070, China.

ABSTRACT

Background: The aim of this study was to investigate the effect of C-reactive protein/oxidised low-density lipoprotein/β2-glycoprotein I (CRP/oxLDL/β2GPI) complex on atherosclerosis (AS) in diabetic BALB/c mice.

Methods: BALB/c mice were fed high-fat and normal diet. Eight weeks later, the mice fed with high-fat diet were injected with streptozotocin (STZ) to induce diabetes. The diabetic mice were respectively injected twice monthly with 20 μg oxLDL, 20 μg β2GPI, 40 μg oxLDL/β2GPI complex, 44 μg CRP/oxLDL/β2GPI complex, and PBS. Aortas were stained with Sudan IV to investigate lipid plaque formation. The infiltration condition of smooth muscle cells (SMCs), macrophages, and T cells in the aortas were determined by immunohistochemistry (IH). The mRNA expressions of receptors associated with lipid metabolism were quantified by real-time PCR. The phosphorylation of p38 mitogen-activated protein kinase (p38MAPK) and MKK3/6 in aorta tissues were assessed by Western blot. The expression of inflammation cytokines was evaluated by protein chip.

Results: The lipid plaques were more extensive, the lumen area was obviously narrower, the ratio of intima and media thickness were increased, and the normal internal elastic lamia structure and endothelial cell disappeared (P < 0.05) in the oxLDL and CRP/oxLDL/β2GPI groups (P < 0.05). CRP/oxLDL/β2GPI complex dramatically promoted infiltration of SMCs, macrophages, and T cells, improved the mRNA expression of ABCA1 and ABCG1, but reduced the mRNA expression of SR-BI and CD36 and increased the phosphorylation of p38MAPK and MKK3/6 (all P < 0.05). The highest expression levels of IL-1, IL-9, PF-4, bFGF, and IGF-II were detected in the CRP/oxLDL/β2GPI group (P < 0.05).

Conclusions: CRP/oxLDL/β2GPI complex aggravated AS in diabetic BALB/c mice by increasing lipid uptake, the mechanism of which may be mediated by the p38MAPK signal pathway.

Show MeSH
Related in: MedlinePlus