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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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Effect of CRP/oxLDL/β2GPI on receptors associated with lipid metabolism gene expression in the aortas. After induction of diabetes and treatment with oxLDL, β2GPI, oxLDL/β2GPI complex, CRP/oxLDL/β2GPI complex and PBS (as described in the Methods section), aortas were removed and the expression of selected genes was measured using real-time PCR. Estimates of the quantity of the PCR product were obtained by densitometry using the Quantity One analysis software package. Results are show as means ± SEM of mRNA level normalized to β-actin in the normal (non-diabetes) group. A: ABCA1 mRNA and ABCG1 mRNA expression level in the aortas. B: CD36 mRNA and SR-BI mRNA expression level in the aortas. Expression level of NC group was normalized as 1, n= 4. *P < 0.05, compared with the CRP/oxLDL/β2GPI group, †P < 0.05, compared with the PBS group, ‡P< 0.05, compared with the NC group.
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Figure 5: Effect of CRP/oxLDL/β2GPI on receptors associated with lipid metabolism gene expression in the aortas. After induction of diabetes and treatment with oxLDL, β2GPI, oxLDL/β2GPI complex, CRP/oxLDL/β2GPI complex and PBS (as described in the Methods section), aortas were removed and the expression of selected genes was measured using real-time PCR. Estimates of the quantity of the PCR product were obtained by densitometry using the Quantity One analysis software package. Results are show as means ± SEM of mRNA level normalized to β-actin in the normal (non-diabetes) group. A: ABCA1 mRNA and ABCG1 mRNA expression level in the aortas. B: CD36 mRNA and SR-BI mRNA expression level in the aortas. Expression level of NC group was normalized as 1, n= 4. *P < 0.05, compared with the CRP/oxLDL/β2GPI group, †P < 0.05, compared with the PBS group, ‡P< 0.05, compared with the NC group.

Mentions: Real-time PCR results showed that the PBS and NC groups seemed to share the same relative expression level of all the target mRNAs. The other DM groups presented the opposite tendency, with significantly up-regulated mRNA level of ATP-binding cassette transporter protein A1 (ABCA1) and ATP-binding cassette transporter protein G1 (ABCG1) (Figure 5A) and down-regulated CD36 mRNA expression level (Figure 5B). The CRP/oxLDL/β2GPI group obviously promoted ABCG-1 mRNA expression and suppressed CD36 mRNA expression compared with the oxLDL group (P<0.05). However, scavenger receptor type B1 (SR-BI) mRNA expression of the CRP/oxLDL/β2GPI group was almost the same as that of the PBS and NC groups, with remarkably depressed SR-BI compared with the other DM groups (P < 0.05) (Figure 5B).


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)

Effect of CRP/oxLDL/β2GPI on receptors associated with lipid metabolism gene expression in the aortas. After induction of diabetes and treatment with oxLDL, β2GPI, oxLDL/β2GPI complex, CRP/oxLDL/β2GPI complex and PBS (as described in the Methods section), aortas were removed and the expression of selected genes was measured using real-time PCR. Estimates of the quantity of the PCR product were obtained by densitometry using the Quantity One analysis software package. Results are show as means ± SEM of mRNA level normalized to β-actin in the normal (non-diabetes) group. A: ABCA1 mRNA and ABCG1 mRNA expression level in the aortas. B: CD36 mRNA and SR-BI mRNA expression level in the aortas. Expression level of NC group was normalized as 1, n= 4. *P < 0.05, compared with the CRP/oxLDL/β2GPI group, †P < 0.05, compared with the PBS group, ‡P< 0.05, compared with the NC group.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 5: Effect of CRP/oxLDL/β2GPI on receptors associated with lipid metabolism gene expression in the aortas. After induction of diabetes and treatment with oxLDL, β2GPI, oxLDL/β2GPI complex, CRP/oxLDL/β2GPI complex and PBS (as described in the Methods section), aortas were removed and the expression of selected genes was measured using real-time PCR. Estimates of the quantity of the PCR product were obtained by densitometry using the Quantity One analysis software package. Results are show as means ± SEM of mRNA level normalized to β-actin in the normal (non-diabetes) group. A: ABCA1 mRNA and ABCG1 mRNA expression level in the aortas. B: CD36 mRNA and SR-BI mRNA expression level in the aortas. Expression level of NC group was normalized as 1, n= 4. *P < 0.05, compared with the CRP/oxLDL/β2GPI group, †P < 0.05, compared with the PBS group, ‡P< 0.05, compared with the NC group.
Mentions: Real-time PCR results showed that the PBS and NC groups seemed to share the same relative expression level of all the target mRNAs. The other DM groups presented the opposite tendency, with significantly up-regulated mRNA level of ATP-binding cassette transporter protein A1 (ABCA1) and ATP-binding cassette transporter protein G1 (ABCG1) (Figure 5A) and down-regulated CD36 mRNA expression level (Figure 5B). The CRP/oxLDL/β2GPI group obviously promoted ABCG-1 mRNA expression and suppressed CD36 mRNA expression compared with the oxLDL group (P<0.05). However, scavenger receptor type B1 (SR-BI) mRNA expression of the CRP/oxLDL/β2GPI group was almost the same as that of the PBS and NC groups, with remarkably depressed SR-BI compared with the other DM groups (P < 0.05) (Figure 5B).

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