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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

Reduction in infarct size and improvement of neurological function by minocycline treatment in the wild type, but not in MCPIP1-deficient mice. The brain infarct size was assessed 48 h after MCAO. (A) Infarct images obtained by TTC staining at 48 h after MCAO. The normal tissue was stained deep red and the infarct was stained milky. (B) Brain infarcts were quantified as percentage area of ischemic hemisphere. The infarct size of minocycline-pretreated wild type mice was significantly reduced compared to that of the control. There was no significant difference in brain infarct size between the minocycline-pretreated and control in MCPIP1-deficient mice. (C) Brain water content as a measure of brain edema of the ischemic hemisphere. The brain edema was significantly reduced at 48 h after MCAO in minocycline-pretreated wild type mice compared to that of the control group. In the MCPIP1-deficient mice, there was no significant difference in brain water content between minocycline-pretreated and control group without minocycline treatment. (D) Neurological function assessment was performed 24 h after MCAO. The neurological scores of minocycline-pretreated wild type mice were significantly improved compared to that of the control. In MCPIP1-deficient mice, there was no significant difference in neurological deficits between minocycline-pretreated and control group. Values represent mean ± SD, *P < 0.05, n = 10 mice per group. MCAO, middle cerebral artery occlusion; MCPIP1, monocyte chemotactic protein-induced protein 1; TTC, 2,3,5-triphenyltetrazolium chloride.
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Fig3: Reduction in infarct size and improvement of neurological function by minocycline treatment in the wild type, but not in MCPIP1-deficient mice. The brain infarct size was assessed 48 h after MCAO. (A) Infarct images obtained by TTC staining at 48 h after MCAO. The normal tissue was stained deep red and the infarct was stained milky. (B) Brain infarcts were quantified as percentage area of ischemic hemisphere. The infarct size of minocycline-pretreated wild type mice was significantly reduced compared to that of the control. There was no significant difference in brain infarct size between the minocycline-pretreated and control in MCPIP1-deficient mice. (C) Brain water content as a measure of brain edema of the ischemic hemisphere. The brain edema was significantly reduced at 48 h after MCAO in minocycline-pretreated wild type mice compared to that of the control group. In the MCPIP1-deficient mice, there was no significant difference in brain water content between minocycline-pretreated and control group without minocycline treatment. (D) Neurological function assessment was performed 24 h after MCAO. The neurological scores of minocycline-pretreated wild type mice were significantly improved compared to that of the control. In MCPIP1-deficient mice, there was no significant difference in neurological deficits between minocycline-pretreated and control group. Values represent mean ± SD, *P < 0.05, n = 10 mice per group. MCAO, middle cerebral artery occlusion; MCPIP1, monocyte chemotactic protein-induced protein 1; TTC, 2,3,5-triphenyltetrazolium chloride.

Mentions: We examined the effects of minocycline treatment on ischemic brain infarction. MCPIP1-deficient and wild type mice were pretreated with minocycline as described in the Methods section, and these mice were subjected to MCAO 12 h after minocycline treatment. The brain infarct size was assessed with TTC staining 48 h after MCAO, and the results showed that the infarct size of minocycline-pretreated wild type mice was significantly reduced compared to that of the control (39.4% ± 5.3% versus 19.3% ± 4.1%, respectively, Figure 3A). MCPIP1-deficient mice failed to evoke minocycline-treatment-induced tolerance compared with that of the control MCPIP1 knockout group without minocycline treatment (54.3% ± 6.5% versus 47.6% ± 6.9%, respectively, Figure 3A). There was no significant difference in brain infarct size between minocycline-pretreated and control in MCPIP1 knockout mice. The neurological functions of mice were determined, and the results showed that the neurological scores of minocycline-pretreated wild type mice were significantly improved compared to that of the control. In MCPIP1-deficient mice, there was no significant difference in neurological deficits between the minocycline-pretreated and control group without minocycline treatment (Figure 3B). Edema is one of the earliest pathological changes after ischemic neuronal damage. The results showed that the brain edema was significantly reduced at 48 h after MCAO in minocycline-pretreated wild type mice compared to that of the control group. In the MCPIP1-deficient mice, there was no significant difference in brain water content between the minocycline-pretreated and control group without minocycline treatment (Figure 3C). We also examined the effects of minocycline treatment post-stroke on ischemic brain infarction and neurological functions. MCPIP1-deficient and wild type mice were treated with minocycline (50 mg/kg per day) at 2 h after MCAO. The results showed that the infarct size of minocycline-treated wild type mice was significantly reduced compared to that of the control (40.3% ± 6.1% versus 23.3% ± 4.7%, P < 0.05, Figure 4A). MCPIP1-deficient mice failed to evoke minocycline-treatment-induced neuroprotection compared with that of the control MCPIP1 knockout group without minocycline treatment (55.1% ± 4.9% versus 52.8% ± 5.3%, P > 0.05, Figure 4A). There was no significant difference in brain infarct size between the minocycline-treated and control in MCPIP1 knockout mice. The neurological functions of mice were determined, and the results showed that the neurological scores of minocycline-treated wild type mice were significantly improved compared to that of the control. In MCPIP1-deficient mice, there was no significant difference in neurological deficits between the minocycline-treated and control group without minocycline treatment (Figure 4B). These results indicated that MCPIP1 may mediate minocycline-induced neuroprotection afforded by minocycline administration after stroke or before stroke.Figure 3


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)

Reduction in infarct size and improvement of neurological function by minocycline treatment in the wild type, but not in MCPIP1-deficient mice. The brain infarct size was assessed 48 h after MCAO. (A) Infarct images obtained by TTC staining at 48 h after MCAO. The normal tissue was stained deep red and the infarct was stained milky. (B) Brain infarcts were quantified as percentage area of ischemic hemisphere. The infarct size of minocycline-pretreated wild type mice was significantly reduced compared to that of the control. There was no significant difference in brain infarct size between the minocycline-pretreated and control in MCPIP1-deficient mice. (C) Brain water content as a measure of brain edema of the ischemic hemisphere. The brain edema was significantly reduced at 48 h after MCAO in minocycline-pretreated wild type mice compared to that of the control group. In the MCPIP1-deficient mice, there was no significant difference in brain water content between minocycline-pretreated and control group without minocycline treatment. (D) Neurological function assessment was performed 24 h after MCAO. The neurological scores of minocycline-pretreated wild type mice were significantly improved compared to that of the control. In MCPIP1-deficient mice, there was no significant difference in neurological deficits between minocycline-pretreated and control group. Values represent mean ± SD, *P < 0.05, n = 10 mice per group. MCAO, middle cerebral artery occlusion; MCPIP1, monocyte chemotactic protein-induced protein 1; TTC, 2,3,5-triphenyltetrazolium chloride.
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Fig3: Reduction in infarct size and improvement of neurological function by minocycline treatment in the wild type, but not in MCPIP1-deficient mice. The brain infarct size was assessed 48 h after MCAO. (A) Infarct images obtained by TTC staining at 48 h after MCAO. The normal tissue was stained deep red and the infarct was stained milky. (B) Brain infarcts were quantified as percentage area of ischemic hemisphere. The infarct size of minocycline-pretreated wild type mice was significantly reduced compared to that of the control. There was no significant difference in brain infarct size between the minocycline-pretreated and control in MCPIP1-deficient mice. (C) Brain water content as a measure of brain edema of the ischemic hemisphere. The brain edema was significantly reduced at 48 h after MCAO in minocycline-pretreated wild type mice compared to that of the control group. In the MCPIP1-deficient mice, there was no significant difference in brain water content between minocycline-pretreated and control group without minocycline treatment. (D) Neurological function assessment was performed 24 h after MCAO. The neurological scores of minocycline-pretreated wild type mice were significantly improved compared to that of the control. In MCPIP1-deficient mice, there was no significant difference in neurological deficits between minocycline-pretreated and control group. Values represent mean ± SD, *P < 0.05, n = 10 mice per group. MCAO, middle cerebral artery occlusion; MCPIP1, monocyte chemotactic protein-induced protein 1; TTC, 2,3,5-triphenyltetrazolium chloride.
Mentions: We examined the effects of minocycline treatment on ischemic brain infarction. MCPIP1-deficient and wild type mice were pretreated with minocycline as described in the Methods section, and these mice were subjected to MCAO 12 h after minocycline treatment. The brain infarct size was assessed with TTC staining 48 h after MCAO, and the results showed that the infarct size of minocycline-pretreated wild type mice was significantly reduced compared to that of the control (39.4% ± 5.3% versus 19.3% ± 4.1%, respectively, Figure 3A). MCPIP1-deficient mice failed to evoke minocycline-treatment-induced tolerance compared with that of the control MCPIP1 knockout group without minocycline treatment (54.3% ± 6.5% versus 47.6% ± 6.9%, respectively, Figure 3A). There was no significant difference in brain infarct size between minocycline-pretreated and control in MCPIP1 knockout mice. The neurological functions of mice were determined, and the results showed that the neurological scores of minocycline-pretreated wild type mice were significantly improved compared to that of the control. In MCPIP1-deficient mice, there was no significant difference in neurological deficits between the minocycline-pretreated and control group without minocycline treatment (Figure 3B). Edema is one of the earliest pathological changes after ischemic neuronal damage. The results showed that the brain edema was significantly reduced at 48 h after MCAO in minocycline-pretreated wild type mice compared to that of the control group. In the MCPIP1-deficient mice, there was no significant difference in brain water content between the minocycline-pretreated and control group without minocycline treatment (Figure 3C). We also examined the effects of minocycline treatment post-stroke on ischemic brain infarction and neurological functions. MCPIP1-deficient and wild type mice were treated with minocycline (50 mg/kg per day) at 2 h after MCAO. The results showed that the infarct size of minocycline-treated wild type mice was significantly reduced compared to that of the control (40.3% ± 6.1% versus 23.3% ± 4.7%, P < 0.05, Figure 4A). MCPIP1-deficient mice failed to evoke minocycline-treatment-induced neuroprotection compared with that of the control MCPIP1 knockout group without minocycline treatment (55.1% ± 4.9% versus 52.8% ± 5.3%, P > 0.05, Figure 4A). There was no significant difference in brain infarct size between the minocycline-treated and control in MCPIP1 knockout mice. The neurological functions of mice were determined, and the results showed that the neurological scores of minocycline-treated wild type mice were significantly improved compared to that of the control. In MCPIP1-deficient mice, there was no significant difference in neurological deficits between the minocycline-treated and control group without minocycline treatment (Figure 4B). These results indicated that MCPIP1 may mediate minocycline-induced neuroprotection afforded by minocycline administration after stroke or before stroke.Figure 3

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