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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

MCPIP1 is expressed in neurons and microglia. Co-localization of MCPIP1 expression (A, D, G) of NSE (B), CD11b (E), and GFAP (H) at 24 h after minocycline treatment. Yellow fluorescence indicates co-localization of MCPIP1/NSE (C) and MCPIP1/CD11b (F). MCPIP1/GFAP (I) showed no yellow-fluorescent structures. n = 5 mice per group. GFAP, glial fibrillary acidic protein; MCPIP1, monocyte chemotactic protein-induced protein 1; NSE, neuron-specific enolase.
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Fig2: MCPIP1 is expressed in neurons and microglia. Co-localization of MCPIP1 expression (A, D, G) of NSE (B), CD11b (E), and GFAP (H) at 24 h after minocycline treatment. Yellow fluorescence indicates co-localization of MCPIP1/NSE (C) and MCPIP1/CD11b (F). MCPIP1/GFAP (I) showed no yellow-fluorescent structures. n = 5 mice per group. GFAP, glial fibrillary acidic protein; MCPIP1, monocyte chemotactic protein-induced protein 1; NSE, neuron-specific enolase.

Mentions: In ischemic stroke, minocycline treatment has been demonstrated to have a significant protective effect against brain damage, but the underlying mechanism remains poorly understood. First, we examined whether minocycline treatment induces MCPIP1 in mouse brain. The MCPIP1 mRNA level in mouse cortex brain was significantly induced by minocycline treatment compared to controls. Significant increase of MCPIP1 transcript level was detected at 6 h and reached 11.5 ± 1.9-fold at 12 h after minocycline treatment (P < 0.001) and began to decline by 24 hr (Figure 1A). Consistently, the MCPIP1 protein level in mouse brain was significantly elevated by minocycline treatment than in the controls: 6.1 ± 0.93-fold increase at 24 h after minocycline treatment and remained high till 48 hr (P < 0.05; Figure 1B). To determine the cellular localization of MCPIP1 expression, sections from mice brain at 24 h after minocycline treatment were subjected to immunohistochemistry with antibody against MCPIP1. MCPIP1 expression was elevated by minocycline treatment and the immunoreactivity co-localized with NSE, a neuron specific marker, implying that the elevated level of MCPIP1 protein induced by minocycline was located mainly in neurons (Figure 2C). Examination of the co-localization of MCPIP1 with CD11b, the leukocyte-specific receptor and regarded as a marker for macrophages/microglia, showed a few MCPIP1/CD11b-positive cells (Figure 2F). Examination of the co-localization study of MCPIP1 with GFAP, a marker for activated astrocytes, showed that MCPIP1 expression was not located in the astrocytes (Figure 2I). These observations indicate that MCPIP1 protein is upregulated mainly in neurons and microglia in the mice brain after minocycline treatment.Figure 1


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)

MCPIP1 is expressed in neurons and microglia. Co-localization of MCPIP1 expression (A, D, G) of NSE (B), CD11b (E), and GFAP (H) at 24 h after minocycline treatment. Yellow fluorescence indicates co-localization of MCPIP1/NSE (C) and MCPIP1/CD11b (F). MCPIP1/GFAP (I) showed no yellow-fluorescent structures. n = 5 mice per group. GFAP, glial fibrillary acidic protein; MCPIP1, monocyte chemotactic protein-induced protein 1; NSE, neuron-specific enolase.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Fig2: MCPIP1 is expressed in neurons and microglia. Co-localization of MCPIP1 expression (A, D, G) of NSE (B), CD11b (E), and GFAP (H) at 24 h after minocycline treatment. Yellow fluorescence indicates co-localization of MCPIP1/NSE (C) and MCPIP1/CD11b (F). MCPIP1/GFAP (I) showed no yellow-fluorescent structures. n = 5 mice per group. GFAP, glial fibrillary acidic protein; MCPIP1, monocyte chemotactic protein-induced protein 1; NSE, neuron-specific enolase.
Mentions: In ischemic stroke, minocycline treatment has been demonstrated to have a significant protective effect against brain damage, but the underlying mechanism remains poorly understood. First, we examined whether minocycline treatment induces MCPIP1 in mouse brain. The MCPIP1 mRNA level in mouse cortex brain was significantly induced by minocycline treatment compared to controls. Significant increase of MCPIP1 transcript level was detected at 6 h and reached 11.5 ± 1.9-fold at 12 h after minocycline treatment (P < 0.001) and began to decline by 24 hr (Figure 1A). Consistently, the MCPIP1 protein level in mouse brain was significantly elevated by minocycline treatment than in the controls: 6.1 ± 0.93-fold increase at 24 h after minocycline treatment and remained high till 48 hr (P < 0.05; Figure 1B). To determine the cellular localization of MCPIP1 expression, sections from mice brain at 24 h after minocycline treatment were subjected to immunohistochemistry with antibody against MCPIP1. MCPIP1 expression was elevated by minocycline treatment and the immunoreactivity co-localized with NSE, a neuron specific marker, implying that the elevated level of MCPIP1 protein induced by minocycline was located mainly in neurons (Figure 2C). Examination of the co-localization of MCPIP1 with CD11b, the leukocyte-specific receptor and regarded as a marker for macrophages/microglia, showed a few MCPIP1/CD11b-positive cells (Figure 2F). Examination of the co-localization study of MCPIP1 with GFAP, a marker for activated astrocytes, showed that MCPIP1 expression was not located in the astrocytes (Figure 2I). These observations indicate that MCPIP1 protein is upregulated mainly in neurons and microglia in the mice brain after minocycline treatment.Figure 1

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