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MCP-induced protein 1 mediates the minocycline-induced neuroprotection against cerebral ischemia/reperfusion injury in vitro and in vivo.

Jin Z, Liang J, Wang J, Kolattukudy PE - J Neuroinflammation (2015)

Bottom Line: Minocycline, a broad-spectrum tetracycline antibiotic, has shown anti-inflammatory and neuroprotective effects in ischemic brain injury.Similarly, in vitro data showed that minocycline significantly induced the expression of MCPIP1 in primary neuron-glial cells, cortical neurons, and reduced oxygen glucose deprivation (OGD)-induced cell death.Our in vitro and in vivo studies demonstrate that MCPIP1 is an important mediator of minocycline-induced protection from brain ischemia.

View Article: PubMed Central - PubMed

Affiliation: School of Basic Medicine, Zhejiang Chinese Medical University, Hangzhou, 310053, Zhejiang, China. jinzq@hotmail.com.

ABSTRACT

Background: Minocycline, a broad-spectrum tetracycline antibiotic, has shown anti-inflammatory and neuroprotective effects in ischemic brain injury. The present study seeks to determine whether monocyte chemotactic protein-induced protein 1 (MCPIP1), a recently identified modulator of inflammatory reactions, is involved in the cerebral neuroprotection conferred by minocycline treatment in the animal model of focal cerebral ischemia and to elucidate the mechanisms of minocycline-induced ischemic brain tolerance.

Methods: Focal cerebral ischemia was induced by middle cerebral artery occlusion (MCAO) for 2 h in male C57BL/6 mice and MCPIP1 knockout mice followed by 24- or 48-h reperfusion. Twelve hours before ischemia or 2 h after MCAO, mice were injected intraperitoneally with 90 mg/kg of minocycline hydrochloride. Thereafter, the animals were injected twice a day, at a dose of 90 mg/kg after ischemia until sacrificed. Transcription and expression of MCPIP1 gene was monitored by quantitative real-time PCR (qRT-PCR), Western blot, and immunohistochemistry. The neurobehavioral scores, infarction volumes, and proinflammatory cytokines in brain and NF-κB signaling were evaluated after ischemia/reperfusion.

Results: MCPIP1 protein and mRNA levels significantly increased in mouse brain undergoing minocycline pretreatment. Minocycline treatment significantly attenuated the infarct volume, neurological deficits, and upregulation of proinflammatory cytokines in the brain of wild type mice after MCAO. MCPIP1-deficient mice failed to evoke minocycline-treatment-induced tolerance compared with that of the control MCPIP1-deficient group without minocycline treatment. Similarly, in vitro data showed that minocycline significantly induced the expression of MCPIP1 in primary neuron-glial cells, cortical neurons, and reduced oxygen glucose deprivation (OGD)-induced cell death. The absence of MCPIP1 blocked minocycline-induced protection on neuron-glial cells and cortical neurons treated with OGD.

Conclusions: Our in vitro and in vivo studies demonstrate that MCPIP1 is an important mediator of minocycline-induced protection from brain ischemia.

No MeSH data available.


Related in: MedlinePlus

Inhibition of NF-κB activation in the ischemic brain by minocycline treatment in the wild type, but not in MCPIP1-deficient mice. A representative Western blot shows protein levels of p-65 phosphorylation. The phosphorylation of p-65 was significantly reduced at 24 h after MCAO in minocycline-pretreated wild type mice compared to that of the control. In MCPIP1-deficient mice, there was no significant difference in p-65 phosphorylation level between the minocycline-pretreated and control group without minocycline treatment. Densitometric analysis was used to quantify phospho-p-65 protein levels versus total p-65 in three independent Western blots, and the data are expressed as the normalized folds with respect to sham. Values represent mean ± SD. MCAO, middle cerebral artery occlusion; MCPIP1, monocyte chemotactic protein-induced protein 1.
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Fig8: Inhibition of NF-κB activation in the ischemic brain by minocycline treatment in the wild type, but not in MCPIP1-deficient mice. A representative Western blot shows protein levels of p-65 phosphorylation. The phosphorylation of p-65 was significantly reduced at 24 h after MCAO in minocycline-pretreated wild type mice compared to that of the control. In MCPIP1-deficient mice, there was no significant difference in p-65 phosphorylation level between the minocycline-pretreated and control group without minocycline treatment. Densitometric analysis was used to quantify phospho-p-65 protein levels versus total p-65 in three independent Western blots, and the data are expressed as the normalized folds with respect to sham. Values represent mean ± SD. MCAO, middle cerebral artery occlusion; MCPIP1, monocyte chemotactic protein-induced protein 1.

Mentions: Since activation of the NF-κB-signaling pathway is involved in the production of proinflammatory cytokines, we tested whether minocycline treatment affects NF-κB activation. The results showed that phosphorylation of p-65 was significantly reduced at 24 h after MCAO in the minocycline-pretreated wild type mice compared to that of the control. In MCPIP1-deficient mice, there was no significant difference in p-65 phosphorylation level between the minocycline-pretreated and control group without minocycline treatment (Figure 8).Figure 8


MCP-induced protein 1 mediates the minocycline-induced neuroprotection against cerebral ischemia/reperfusion injury in vitro and in vivo.

Jin Z, Liang J, Wang J, Kolattukudy PE - J Neuroinflammation (2015)

Inhibition of NF-κB activation in the ischemic brain by minocycline treatment in the wild type, but not in MCPIP1-deficient mice. A representative Western blot shows protein levels of p-65 phosphorylation. The phosphorylation of p-65 was significantly reduced at 24 h after MCAO in minocycline-pretreated wild type mice compared to that of the control. In MCPIP1-deficient mice, there was no significant difference in p-65 phosphorylation level between the minocycline-pretreated and control group without minocycline treatment. Densitometric analysis was used to quantify phospho-p-65 protein levels versus total p-65 in three independent Western blots, and the data are expressed as the normalized folds with respect to sham. Values represent mean ± SD. MCAO, middle cerebral artery occlusion; MCPIP1, monocyte chemotactic protein-induced protein 1.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4359584&req=5

Fig8: Inhibition of NF-κB activation in the ischemic brain by minocycline treatment in the wild type, but not in MCPIP1-deficient mice. A representative Western blot shows protein levels of p-65 phosphorylation. The phosphorylation of p-65 was significantly reduced at 24 h after MCAO in minocycline-pretreated wild type mice compared to that of the control. In MCPIP1-deficient mice, there was no significant difference in p-65 phosphorylation level between the minocycline-pretreated and control group without minocycline treatment. Densitometric analysis was used to quantify phospho-p-65 protein levels versus total p-65 in three independent Western blots, and the data are expressed as the normalized folds with respect to sham. Values represent mean ± SD. MCAO, middle cerebral artery occlusion; MCPIP1, monocyte chemotactic protein-induced protein 1.
Mentions: Since activation of the NF-κB-signaling pathway is involved in the production of proinflammatory cytokines, we tested whether minocycline treatment affects NF-κB activation. The results showed that phosphorylation of p-65 was significantly reduced at 24 h after MCAO in the minocycline-pretreated wild type mice compared to that of the control. In MCPIP1-deficient mice, there was no significant difference in p-65 phosphorylation level between the minocycline-pretreated and control group without minocycline treatment (Figure 8).Figure 8

Bottom Line: Minocycline, a broad-spectrum tetracycline antibiotic, has shown anti-inflammatory and neuroprotective effects in ischemic brain injury.Similarly, in vitro data showed that minocycline significantly induced the expression of MCPIP1 in primary neuron-glial cells, cortical neurons, and reduced oxygen glucose deprivation (OGD)-induced cell death.Our in vitro and in vivo studies demonstrate that MCPIP1 is an important mediator of minocycline-induced protection from brain ischemia.

View Article: PubMed Central - PubMed

Affiliation: School of Basic Medicine, Zhejiang Chinese Medical University, Hangzhou, 310053, Zhejiang, China. jinzq@hotmail.com.

ABSTRACT

Background: Minocycline, a broad-spectrum tetracycline antibiotic, has shown anti-inflammatory and neuroprotective effects in ischemic brain injury. The present study seeks to determine whether monocyte chemotactic protein-induced protein 1 (MCPIP1), a recently identified modulator of inflammatory reactions, is involved in the cerebral neuroprotection conferred by minocycline treatment in the animal model of focal cerebral ischemia and to elucidate the mechanisms of minocycline-induced ischemic brain tolerance.

Methods: Focal cerebral ischemia was induced by middle cerebral artery occlusion (MCAO) for 2 h in male C57BL/6 mice and MCPIP1 knockout mice followed by 24- or 48-h reperfusion. Twelve hours before ischemia or 2 h after MCAO, mice were injected intraperitoneally with 90 mg/kg of minocycline hydrochloride. Thereafter, the animals were injected twice a day, at a dose of 90 mg/kg after ischemia until sacrificed. Transcription and expression of MCPIP1 gene was monitored by quantitative real-time PCR (qRT-PCR), Western blot, and immunohistochemistry. The neurobehavioral scores, infarction volumes, and proinflammatory cytokines in brain and NF-κB signaling were evaluated after ischemia/reperfusion.

Results: MCPIP1 protein and mRNA levels significantly increased in mouse brain undergoing minocycline pretreatment. Minocycline treatment significantly attenuated the infarct volume, neurological deficits, and upregulation of proinflammatory cytokines in the brain of wild type mice after MCAO. MCPIP1-deficient mice failed to evoke minocycline-treatment-induced tolerance compared with that of the control MCPIP1-deficient group without minocycline treatment. Similarly, in vitro data showed that minocycline significantly induced the expression of MCPIP1 in primary neuron-glial cells, cortical neurons, and reduced oxygen glucose deprivation (OGD)-induced cell death. The absence of MCPIP1 blocked minocycline-induced protection on neuron-glial cells and cortical neurons treated with OGD.

Conclusions: Our in vitro and in vivo studies demonstrate that MCPIP1 is an important mediator of minocycline-induced protection from brain ischemia.

No MeSH data available.


Related in: MedlinePlus