Neuroprotective activity of leukemia inhibitory factor is relayed through myeloid zinc finger-1 in a rat model of stroke

Objective: To determine whether leukemia inhibitory factor (LIF) upregulates superoxide dismutase 3 (SOD3), a neuroprotective antioxidant enzyme, through the transcription factor myeloid zinc finger-1 (MZF-1). Hypothesis: MZF-1 facilitates LIF-mediated neuroprotection by increasing transcription of the SOD3 gene in neurons. Methods: After middle cerebral artery occlusion or sham surgery, male Sprague-Dawley rats were injected with PBS or LIF (125 μg/kg) (n=4 per group). Rats were euthanized 72 h after injury and western blotting was used to measure MZF-1 protein expression in brain tissue. For in vitro studies, rat neurons were transfected with scrambled or MZF-1 siRNA (n=3 per group). Neurons were treated with PBS or 200 ng/mL LIF prior to 24 h in vitro ischemia induced by oxygen glucose deprivation. Lactate dehydrogenase (LDH) release was measured to assess neuronal death. MZF-1 levels were quantified in cultured neurons with immunocytochemistry. SOD3 and MZF-1 mRNA levels were measured with real-time PCR. Results: LIF (0.938 OD units ± 0.170), but not PBS (0.562 OD units ± 0.223), significantly increased MZF-1 protein expression in ipsilateral tissue 72 h after stroke compared to sham surgery (0.411 OD units ± 0.039, p<0.05). LIF treatment prior to 24 h in vitro ischemia significantly increased the percentage of MZF-1-positive neurons (52.17 % ± 0.93) compared to PBS 44.84 % ± 1.11, p<0.01). PCR results confirmed the increase in MZF-1 mRNA (1.89 fold change ± 0.33) in LIF-treated neurons compared to PBS-treated neurons (1.00 fold change ± 0.23, p<0.05). Moreover, LIF increased SOD3 mRNA after 24 h ischemia (1.35 fold change ± 0.03) compared to PBS (1.00 fold change ± 0.13, p<0.05). LIF decreased LDH release (363,967 ± 68,557 neuronal units) compared to PBS (559,856 ± 60,555 neuronal units, p<0.05) among neurons transfected with scrambled siRNA. However, MZF-1 siRNA attenuated the neuroprotective effect of LIF (690,633 neuronal units ± 19,167; p<0.05). Conclusions: MZF-1 plays a fundamental role in a novel neuroprotective pathway by enhancing SOD3 expression and provides insight regarding protection by antioxidant enzymes during stroke.

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Leukemia inhibitory factor modulates the peripheral immune response in a rat model of emergent large vessel occlusion

Journal of Neuroinflammation ◽

10.1186/s12974-018-1326-y ◽

2018 ◽

Vol 15 (1) ◽

Cited By ~ 6

Author(s):

Stephanie M. Davis ◽

Lisa A. Collier ◽

Edric D. Winford ◽

Christopher C. Leonardo ◽

Craig T. Ajmo ◽

...

Keyword(s):

Immune Response ◽

Rat Model ◽

Leukemia Inhibitory Factor ◽

Large Vessel ◽

Inhibitory Factor ◽

Large Vessel Occlusion ◽

Vessel Occlusion ◽

Peripheral Immune Response

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Evaluation of Leukemia Inhibitory Factor (LIF) in a Rat Model of Postoperative Pain

Journal of Pain ◽

10.1016/j.jpain.2011.02.351 ◽

2011 ◽

Vol 12 (7) ◽

pp. 819-832 ◽

Cited By ~ 8

Author(s):

Christina M. Spofford ◽

Shekher Mohan ◽

Sinyoung Kang ◽

Jun Ho Jang ◽

Timothy J. Brennan

Keyword(s):

Postoperative Pain ◽

Rat Model ◽

Leukemia Inhibitory Factor ◽

Inhibitory Factor

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Zinc finger proteins 36 (ZFP36s) family and leukemia inhibitory factor (LIF) expression in the human endometrial cycle: a prospective case-control study

Fertility and Sterility ◽

10.1016/j.fertnstert.2012.07.301 ◽

2012 ◽

Vol 98 (3) ◽

pp. S83

Author(s):

E. Evans-Hoeker ◽

S.L. Young ◽

L. Yuan ◽

B. Lessey ◽

K. Caron ◽

...

Keyword(s):

Zinc Finger ◽

Leukemia Inhibitory Factor ◽

Case Control Study ◽

Zinc Finger Proteins ◽

Case Control ◽

Inhibitory Factor ◽

Control Study ◽

Prospective Case Control Study

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Neural Stem Cells Transfected with Leukemia Inhibitory Factor Promote Neuroprotection in a Rat Model of Cerebral Ischemia

Neuroscience Bulletin ◽

10.1007/s12264-019-00405-5 ◽

2019 ◽

Vol 35 (5) ◽

pp. 901-908 ◽

Cited By ~ 6

Author(s):

Lili Tian ◽

Wenli Zhu ◽

Yuanchu Liu ◽

Ye Gong ◽

Aowei Lv ◽

...

Keyword(s):

Stem Cells ◽

Cerebral Ischemia ◽

Neural Stem Cells ◽

Rat Model ◽

Leukemia Inhibitory Factor ◽

Inhibitory Factor

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Leukemia Inhibitory Factor Decreases Neurogenesis and Angiogenesis in a Rat Model of Intracerebral Hemorrhage

Current Medical Science ◽

10.1007/s11596-019-2034-2 ◽

2019 ◽

Vol 39 (2) ◽

pp. 298-304 ◽

Cited By ~ 1

Author(s):

Chuan-zhen Liu ◽

Hua-jun Zhou ◽

Jian-hua Zhong ◽

Tao Tang ◽

Han-jin Cui ◽

...

Keyword(s):

Intracerebral Hemorrhage ◽

Rat Model ◽

Leukemia Inhibitory Factor ◽

Inhibitory Factor

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Abstract T P248: Leukemia Inhibitory Factor Induced-Akt Activation Protects Neurons from Ischemic Damage by Increasing Superoxide Dismutase Activity and Expression

Stroke ◽

10.1161/str.46.suppl_1.tp248 ◽

2015 ◽

Vol 46 (suppl_1) ◽

Author(s):

Stephanie M Davis ◽

Lisa A Collier ◽

Christopher C Leonardo ◽

Hilary A Seifert ◽

Craig T Ajmo ◽

...

Keyword(s):

Superoxide Dismutase ◽

Lactate Dehydrogenase ◽

Zinc Finger ◽

Leukemia Inhibitory Factor ◽

Glucose Deprivation ◽

Oxygen Glucose Deprivation ◽

Akt Inhibitor ◽

Sod Activity ◽

Myeloid Zinc Finger

Objective: To determine the molecular mechanisms by which leukemia inhibitory factor (LIF) protects neurons during permanent middle cerebral artery occlusion (MCAO). Hypothesis: LIF protects neurons from ischemic oxidative injury through Akt-mediated increases in superoxide dismutase 3 (SOD3) expression and total SOD activity. Methods: Male Sprague-Dawley rats injected with vehicle (PBS) or LIF (125 μg/kg) were euthanized 24, 48, and 72 h post-MCAO or sham surgery for assessment of SOD activity and SOD3 expression. Rat cortical neurons were subjected to 24 h oxygen glucose deprivation or normoxia following treatment with vehicle , 50, 200, or 1000 ng/mL LIF (n=3 per group). In a third experiment, neurons were treated with vehicle, 50 ng/mL LIF,10 μM Akt inhibitor, or 50 ng/mL LIF+10 μM Akt inhibitor (n=3 per group) prior to oxygen glucose deprivation. Lactate dehydrogenase levels in media were measured to assess neuronal death. We used immunocytochemistry to assess expression of SOD3, phospho-Akt (Ser473), and myeloid zinc finger-1 in neurons. Results: LIF significantly increased brain SOD activity (2.085 ± 0.476 U/mg lysate; n=5) compared to vehicle (0.919 ± 0.285 U/mg lysate; n=7) 72 h post-MCAO (p<0.05). LIF significantly increased SOD3 protein expression in the brain (3.707 ± 0.541 units; n=3) compared to vehicle (1.401 ± 0.825 units; n=3) at 72 h post-MCAO (p<0.05). Co-localization of phospho-Akt and SOD3 occurred 24 h post-MCAO. Additionally, 50 ng/mL LIF (0.789 ± 0.018 U/mL media; n=3) significantly decreased lactate dehydrogenase levels in vitro compared to vehicle (1.000 ± 0.023 U/mL media; n=3) (p<0.0001). Co-incubation of 50/mL ng LIF with 10 μM Akt inhibitor (1.070 ±0.106 U/mL media; n=3) reversed the protective effect of LIF (0.705 ± 0.050 U/mL media; n=3) (p<0.01). Increases in SOD3, phospho-Akt, and myeloid zinc finger-1 staining after oxygen glucose deprivation and LIF were abolished upon Akt inhibition. Conclusion: Our data demonstrates that LIF-mediated protection against ischemia in vivo and in vitro results from Akt-dependent increases in SOD3 expression and SOD activity.

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