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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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Histological quantitation of the AS lesion in aortas. After induction of diabetes and treatment with 20 μg oxLDL, 20 μg β2GPI, 40 μg oxLDL/β2GPI complex, 44 μg CRP/oxLDL/β2GPI complex and PBS (as described in the Methods section), aortas were removed, longitudinally cut open,stained with Sudan IV, or paraffin-embedded, sectioned, deparaffinated and then stained with haematoxylin and eosin. The quantitation of the AS lesion and intima thickness were measured using Image Pro-Plus 6.0. The percentage of the plaque area to the total aorta area and the ratios of intima/media thickness of lesions at the aortic roots were calculated. Results are shown as means ± SEM. A: Representative images of Sudan IV stain(4×). B: Percentage of plaque area to total aorta area (n=4 mice per group). *P < 0.05, compared with the CRP/oxLDL/β2GPI group, †P < 0.05, compared with the PBS group. C: Representative H&E stain at the root of the aortas, 200× magnification. D: The ratios of intima/media thickness of lesions at the aortic roots (n=4). *P < 0.05, compared with the CRP/oxLDL/β2GPI group, †P < 0.05, compared with the PBS group.
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Figure 3: Histological quantitation of the AS lesion in aortas. After induction of diabetes and treatment with 20 μg oxLDL, 20 μg β2GPI, 40 μg oxLDL/β2GPI complex, 44 μg CRP/oxLDL/β2GPI complex and PBS (as described in the Methods section), aortas were removed, longitudinally cut open,stained with Sudan IV, or paraffin-embedded, sectioned, deparaffinated and then stained with haematoxylin and eosin. The quantitation of the AS lesion and intima thickness were measured using Image Pro-Plus 6.0. The percentage of the plaque area to the total aorta area and the ratios of intima/media thickness of lesions at the aortic roots were calculated. Results are shown as means ± SEM. A: Representative images of Sudan IV stain(4×). B: Percentage of plaque area to total aorta area (n=4 mice per group). *P < 0.05, compared with the CRP/oxLDL/β2GPI group, †P < 0.05, compared with the PBS group. C: Representative H&E stain at the root of the aortas, 200× magnification. D: The ratios of intima/media thickness of lesions at the aortic roots (n=4). *P < 0.05, compared with the CRP/oxLDL/β2GPI group, †P < 0.05, compared with the PBS group.

Mentions: Pale yellow wax-like hills were spread along the aortic intima of the DM group, which were lined with lipid streak and atherosclerotic plaque. The entire aortic intima was uneven with a stiff wall. Tissues of the DM group seemed crisper and more difficult to extend, except those in the PBS group. Among all the DM groups, PBS group possessed the least lipid deposits, whereas CRP/oxLDL/β2GPI group exhibited relatively obvious AS plaques, ranking second to the oxLDL group (Figure 3A). As seen in Figure 3B, the lesion in the CRP/oxLDL/β2GPI group was about 2.4-fold that of the β2GPI group (P<0.05), 1.66-fold that of the oxLDL/β2GPI group (P<0.05), and 6-fold that of the PBS group (P<0.05).


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)

Histological quantitation of the AS lesion in aortas. After induction of diabetes and treatment with 20 μg oxLDL, 20 μg β2GPI, 40 μg oxLDL/β2GPI complex, 44 μg CRP/oxLDL/β2GPI complex and PBS (as described in the Methods section), aortas were removed, longitudinally cut open,stained with Sudan IV, or paraffin-embedded, sectioned, deparaffinated and then stained with haematoxylin and eosin. The quantitation of the AS lesion and intima thickness were measured using Image Pro-Plus 6.0. The percentage of the plaque area to the total aorta area and the ratios of intima/media thickness of lesions at the aortic roots were calculated. Results are shown as means ± SEM. A: Representative images of Sudan IV stain(4×). B: Percentage of plaque area to total aorta area (n=4 mice per group). *P < 0.05, compared with the CRP/oxLDL/β2GPI group, †P < 0.05, compared with the PBS group. C: Representative H&E stain at the root of the aortas, 200× magnification. D: The ratios of intima/media thickness of lesions at the aortic roots (n=4). *P < 0.05, compared with the CRP/oxLDL/β2GPI group, †P < 0.05, compared with the PBS group.
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Related In: Results  -  Collection

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Figure 3: Histological quantitation of the AS lesion in aortas. After induction of diabetes and treatment with 20 μg oxLDL, 20 μg β2GPI, 40 μg oxLDL/β2GPI complex, 44 μg CRP/oxLDL/β2GPI complex and PBS (as described in the Methods section), aortas were removed, longitudinally cut open,stained with Sudan IV, or paraffin-embedded, sectioned, deparaffinated and then stained with haematoxylin and eosin. The quantitation of the AS lesion and intima thickness were measured using Image Pro-Plus 6.0. The percentage of the plaque area to the total aorta area and the ratios of intima/media thickness of lesions at the aortic roots were calculated. Results are shown as means ± SEM. A: Representative images of Sudan IV stain(4×). B: Percentage of plaque area to total aorta area (n=4 mice per group). *P < 0.05, compared with the CRP/oxLDL/β2GPI group, †P < 0.05, compared with the PBS group. C: Representative H&E stain at the root of the aortas, 200× magnification. D: The ratios of intima/media thickness of lesions at the aortic roots (n=4). *P < 0.05, compared with the CRP/oxLDL/β2GPI group, †P < 0.05, compared with the PBS group.
Mentions: Pale yellow wax-like hills were spread along the aortic intima of the DM group, which were lined with lipid streak and atherosclerotic plaque. The entire aortic intima was uneven with a stiff wall. Tissues of the DM group seemed crisper and more difficult to extend, except those in the PBS group. Among all the DM groups, PBS group possessed the least lipid deposits, whereas CRP/oxLDL/β2GPI group exhibited relatively obvious AS plaques, ranking second to the oxLDL group (Figure 3A). As seen in Figure 3B, the lesion in the CRP/oxLDL/β2GPI group was about 2.4-fold that of the β2GPI group (P<0.05), 1.66-fold that of the oxLDL/β2GPI group (P<0.05), and 6-fold that of the PBS group (P<0.05).

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