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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

Minocycline neuroprotection to OGD-induced neuronal damage via MCPIP1 in neuron-glia cells from MCPIP1−/−mice. (A, B) Cell viability assays by MTT and cell death rate by trypan blue inclusion revealed that treatment of OGD on the mixed neuron-glia cells resulted in cell death and pretreatment with minocycline can increase the resistance to OGD-induced neuronal damage. Conversely, there was no significant difference on OGD-induced neuronal damage in neuron-glia cells from MCPIP1−/− mice with the treatment of minocycline compared with that of the control group without minocycline treatment. (C, D) Pretreatment of minocycline decreased OGD-induced expression of TNFα and IL-1β in the mixed neuron-glia cells. There was no significant reduction of OGD-induced TNFα and IL-1β expression in mixed neuron-glia cells from MCPIP1−/− mice with the treatment of minocycline compared with that of the control group without minocycline treatment. MCPIP1, monocyte chemotactic protein-induced protein 1; MTT, 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide; OGD, oxygen glucose deprivation.
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Fig6: Minocycline neuroprotection to OGD-induced neuronal damage via MCPIP1 in neuron-glia cells from MCPIP1−/−mice. (A, B) Cell viability assays by MTT and cell death rate by trypan blue inclusion revealed that treatment of OGD on the mixed neuron-glia cells resulted in cell death and pretreatment with minocycline can increase the resistance to OGD-induced neuronal damage. Conversely, there was no significant difference on OGD-induced neuronal damage in neuron-glia cells from MCPIP1−/− mice with the treatment of minocycline compared with that of the control group without minocycline treatment. (C, D) Pretreatment of minocycline decreased OGD-induced expression of TNFα and IL-1β in the mixed neuron-glia cells. There was no significant reduction of OGD-induced TNFα and IL-1β expression in mixed neuron-glia cells from MCPIP1−/− mice with the treatment of minocycline compared with that of the control group without minocycline treatment. MCPIP1, monocyte chemotactic protein-induced protein 1; MTT, 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide; OGD, oxygen glucose deprivation.

Mentions: Given the observed effects of minocycline on wild type and MCPIP1−/− mice subjected to MCAO, we investigated the roles of minocycline on OGD-induced neuronal damage in primary neuron-glia cells from wild type and MCPIP1−/− mice. Cell viability assays revealed that the treatment of OGD on the mixed neuron-glia cells resulted in cell death, and pretreatment with minocycline (10 μM) increased the resistance to OGD-induced neuronal damage (Figure 6A, B) and reduced the expression of TNFα and IL-1β induced by OGD in neuron-glia cells (Figure 6C, D). More significantly, cell viability analysis revealed that depletion of MCPIP1 in neuron-glia cells from MCPIP1−/− mice reduced the protective effects of minocycline on the cells subjected to OGD. In addition, MCPIP1 depletion in neuron-glia cells also increased the expression of TNFα and IL-1β induced by OGD in the cells pretreated by minocycline compared with that of the control group from the wild type. We also investigated the roles of minocycline on OGD-induced neuronal damage in primary cortical neurons from wild type mice with or without knockdown of MCPIP1. Minocycline (10 μM) significantly induced the expression of MCPIP1 in cortical neurons (Figure 7A). Cell viability assays revealed that the treatment of OGD on cortical neurons resulted in cell death, and the pretreatment with minocycline can increase the resistance to OGD-induced neuronal damage (Figure 7B) and reduced the expression of TNFα induced by OGD in cortical neurons (Figure 7C). More interestingly, cell viability analysis revealed that the depletion of MCPIP1 by siMCPIP1 reduced the protective effects of minocycline on cortical neurons subjected to OGD (Figure 7B). In addition, MCPIP1 depletion significantly increased the expression of TNFα induced by OGD in cortical neurons pretreated by minocycline compared with that of the control group of siControl (Figure 7C).Figure 6


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)

Minocycline neuroprotection to OGD-induced neuronal damage via MCPIP1 in neuron-glia cells from MCPIP1−/−mice. (A, B) Cell viability assays by MTT and cell death rate by trypan blue inclusion revealed that treatment of OGD on the mixed neuron-glia cells resulted in cell death and pretreatment with minocycline can increase the resistance to OGD-induced neuronal damage. Conversely, there was no significant difference on OGD-induced neuronal damage in neuron-glia cells from MCPIP1−/− mice with the treatment of minocycline compared with that of the control group without minocycline treatment. (C, D) Pretreatment of minocycline decreased OGD-induced expression of TNFα and IL-1β in the mixed neuron-glia cells. There was no significant reduction of OGD-induced TNFα and IL-1β expression in mixed neuron-glia cells from MCPIP1−/− mice with the treatment of minocycline compared with that of the control group without minocycline treatment. MCPIP1, monocyte chemotactic protein-induced protein 1; MTT, 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide; OGD, oxygen glucose deprivation.
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Related In: Results  -  Collection

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Fig6: Minocycline neuroprotection to OGD-induced neuronal damage via MCPIP1 in neuron-glia cells from MCPIP1−/−mice. (A, B) Cell viability assays by MTT and cell death rate by trypan blue inclusion revealed that treatment of OGD on the mixed neuron-glia cells resulted in cell death and pretreatment with minocycline can increase the resistance to OGD-induced neuronal damage. Conversely, there was no significant difference on OGD-induced neuronal damage in neuron-glia cells from MCPIP1−/− mice with the treatment of minocycline compared with that of the control group without minocycline treatment. (C, D) Pretreatment of minocycline decreased OGD-induced expression of TNFα and IL-1β in the mixed neuron-glia cells. There was no significant reduction of OGD-induced TNFα and IL-1β expression in mixed neuron-glia cells from MCPIP1−/− mice with the treatment of minocycline compared with that of the control group without minocycline treatment. MCPIP1, monocyte chemotactic protein-induced protein 1; MTT, 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide; OGD, oxygen glucose deprivation.
Mentions: Given the observed effects of minocycline on wild type and MCPIP1−/− mice subjected to MCAO, we investigated the roles of minocycline on OGD-induced neuronal damage in primary neuron-glia cells from wild type and MCPIP1−/− mice. Cell viability assays revealed that the treatment of OGD on the mixed neuron-glia cells resulted in cell death, and pretreatment with minocycline (10 μM) increased the resistance to OGD-induced neuronal damage (Figure 6A, B) and reduced the expression of TNFα and IL-1β induced by OGD in neuron-glia cells (Figure 6C, D). More significantly, cell viability analysis revealed that depletion of MCPIP1 in neuron-glia cells from MCPIP1−/− mice reduced the protective effects of minocycline on the cells subjected to OGD. In addition, MCPIP1 depletion in neuron-glia cells also increased the expression of TNFα and IL-1β induced by OGD in the cells pretreated by minocycline compared with that of the control group from the wild type. We also investigated the roles of minocycline on OGD-induced neuronal damage in primary cortical neurons from wild type mice with or without knockdown of MCPIP1. Minocycline (10 μM) significantly induced the expression of MCPIP1 in cortical neurons (Figure 7A). Cell viability assays revealed that the treatment of OGD on cortical neurons resulted in cell death, and the pretreatment with minocycline can increase the resistance to OGD-induced neuronal damage (Figure 7B) and reduced the expression of TNFα induced by OGD in cortical neurons (Figure 7C). More interestingly, cell viability analysis revealed that the depletion of MCPIP1 by siMCPIP1 reduced the protective effects of minocycline on cortical neurons subjected to OGD (Figure 7B). In addition, MCPIP1 depletion significantly increased the expression of TNFα induced by OGD in cortical neurons pretreated by minocycline compared with that of the control group of siControl (Figure 7C).Figure 6

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