Necrostatin-1 reduces histopathology and improves functional outcome after controlled cortical impact in mice - PubMed (original) (raw)

Necrostatin-1 reduces histopathology and improves functional outcome after controlled cortical impact in mice

Zerong You et al. J Cereb Blood Flow Metab. 2008 Sep.

Abstract

Necroptosis is a newly identified type of programmed necrosis initiated by the activation of tumor necrosis factor alpha (TNFalpha)/Fas. Necrostatin-1 is a specific inhibitor of necroptosis that reduces ischemic tissue damage in experimental stroke models. We previously reported decreased tissue damage and improved functional outcome after controlled cortical impact (CCI) in mice deficient in TNFalpha and Fas. Hence, we hypothesized that necrostatin-1 would reduce histopathology and improve functional outcome after CCI in mice. Compared with vehicle-/inactive analog-treated controls, mice administered necrostatin-1 before CCI had decreased propidium iodide-positive cells in the injured cortex and dentate gyrus (6 h), decreased brain tissue damage (days 14, 35), improved motor (days 1 to 7), and Morris water maze performance (days 8 to 14) after CCI. Improved spatial memory was observed even when drug was administered 15 mins after CCI. Necrostatin-1 treatment did not reduce caspase-3-positive cells in the dentate gyrus or cortex, consistent with a known caspase-independent mechanism of necrostatin-1. However, necrostatin-1 reduced brain neutrophil influx and microglial activation at 48 h, suggesting a novel anti-inflammatory effect in traumatic brain injury (TBI). The data suggest that necroptosis plays a significant role in the pathogenesis of cell death and functional outcome after TBI and that necrostatin-1 may have therapeutic potential for patients with TBI.

PubMed Disclaimer

Figures

Figure 1

Figure 1

Brain lesion volume is reduced in Necrostatin-1 treated mice after controlled cortical impact (CCI). (A) Pretreatment with necrostatin-1 reduced lesion volume at 14 days after CCI compared to treatment with vehicle (Veh) or inactive analogue (ANA)(p < 0.001 ANOVA, *p < 0.05 versus ANA or Veh). Pretreatment with necrostatin-1 also reduced lesion size assessed at 35 days (*p < 0.05). For 14 days, n = 7–16/group; for 35 days, n = 4/group). (B) Necrostatin-1 was administered after CCI at the times indicated. Post injury treatment with necrostatin-1 reduced lesion size compared to vehicle when administered at 5 or 15 min but not 30 min after CCI (*p < 0.05, n = 4–5/group).

Figure 2

Figure 2

Reduced motor deficits after controlled cortical impact (CCI) in Necrostatin-1 treated mice. Vestibulo-motor function assessed by the wire grip test was similar among all groups prior to CCI. Post injury motor function was significantly improved in mice administered necrostatin-1 (Nec-1) compared to inactive analogue (ANA) or vehicle (Veh) (p < 0.05 for group effect, n=8/group). (B) Administration of necrostatin-1 (n = 5) to mice 5 min after CCI did not affect motor performance vs. vehicle treated animals (n = 9).

Figure 3

Figure 3

Necrostatin-1 treatment improves Morris water maze (MWM) performance after controlled cortical impact (CCI). (A) Morris water maze performance in naive adult animals administered necrostatin-1 (Nec-1) or vehicle (Veh). No difference in performance was observed between uninjured Veh- and Nec-1-treated mice in hidden or visible platform trials, or in probe trials (n=4/group). (B) Performance in the hidden platform trials was significantly improved in animals pretreated with Necrostatin-1 versus inactive analogue (ANA) or vehicle (Veh) (p < 0.05 for group effect, n = 7/group). (C) Probe trial performance in mice administered Necrostatin-1 (Nec-1) vs. Veh or ANA. No effect of drug treatment was observed in sham injured mice on probe trial performance, however injured mice administered Nec-1 before or at 5 or 15 minutes after CCI had improved performance vs. vehicle-treated animals (*p < 0.05), n= 4–9/group).

Figure 4

Figure 4

Pre-treatment with necrostatin-1 (Nec-1) reduces propidium iodide (PI)-positive cells at 6 h after controlled cortical impact (CCI) in injured cortex and hippocampus. (A) Representative photomicrographs showing reduced numbers of PI-positive cells in cortical and dentate gyrus brain regions after CCI in Nec-1 and vehicle-treated mice. Magnification × 200. (B) Quantitation of PI-positive cells in injured cortex and dentate gyrus. *p < 0.01 vs. vehicle treated animals (n = 12/group).

Figure 5

Figure 5

No effect of necrostatin-1 on caspase-3-positive cells after controlled cortical impact. Necrostatin-1 or vehicle was administered to mice (n = 7–9/group) immediately before CCI and caspase-3 positive cells were quantitated in injured cortex and dentate gyrus at 48 h. (A) Representative photomicrographs showing similar numbers of caspase-3-positive cells in dentate gyrus after CCI in Necrostatin-1 (Nec-1) and vehicle-treated (Veh) mice. (B) Quantitation of caspase-3-positive cells in injured cortex and dentate gyrus.

Figure 6

Figure 6

Pre-treatment with necrostatin-1 reduces neuroinflammation assessed at 48 h after controlled cortical impact. (A) Representative photomicrographs (top panels) showing neutrophil accumulation in injured cortex in mice administered vehicle (Veh) or necrostatin-1 (Nec-1). Bottom panels show quantitation of neutrophils in injured cortices of Veh and Nec-1 treated mice. * p < 0.05 vs. Veh (n = 10–11/group). (B) Marked reduction in microglial activation in mice administered necrostatin-1 vs. vehicle prior to controlled cortical impact. Upper panels show representative immunohistochemical staining using the microglial specific marker IBA-1. The graph in the lower panel shows quantitation of microglial activation using image analysis software. *p < 0.05 vs. vehicle (n = 7–9/group). Magnification × 200 in all photomicrographs.

Similar articles

Cited by

References

    1. Aoyama N, Katayama Y, Kawamata T, Maeda T, Mori T, Yamamoto T, Kikuchi T, Uwahodo Y. Effects of antioxidant, OPC-14117, on secondary cellular damage and behavioral deficits following cortical contusion in the rat. Brain Res. 2002;934:117–124. - PubMed
    1. Beer R, Franz G, Schopf M, Reindl M, Zelger B, Schmutzhard E, Poewe W, Kampfl A. Expression of Fas and Fas ligand after experimental traumatic brain injury in the rat. J Cereb Blood Flow Metab. 2000;20:669–677. - PubMed
    1. Bermpohl D, You Z, Korsmeyer SJ, Moskowitz MA, Whalen MJ. Traumatic brain injury in mice deficient in Bid: effects on histopathology and functional outcome. J Cereb Blood Flow Metab. 2006;26:625–633. - PubMed
    1. Bermpohl DYZ, Lo EH, Kim H, Moskowitz MA, Whalen MJ. TNF alpha and Fas mediate tissue damage and functional outcome after traumatic brain injury in mice. J Cereb Blood Flow Metab. 2007 In press. - PubMed
    1. Clark RS, Kochanek PM, Watkins SC, Chen M, Dixon CE, Seidberg NA, Melick J, Loeffert JE, Nathaniel PD, Jin KL, Graham SH. Caspase-3 mediated neuronal death after traumatic brain injury in rats. J Neurochem. 2000;74:740–753. - PubMed

Publication types

MeSH terms

Substances

Grants and funding

LinkOut - more resources