TGF-β1抑制大鼠创伤性颅脑损伤后海马神经元的凋亡

目的转化生长因子β1（TGF-β1）在星形胶质细胞增生和神经元的存活方面都发挥这重要作用,但是其在创伤性颅脑损伤（TBI）中的作用还未见报道。方法本课题组先前的研究利用大鼠模型发现在大鼠颅脑损伤后,TGF-β1的表达显著上调,表明TGF-β1参与到创伤性颅脑损伤修复的过程中。结果本研究聚焦在TGF-β1对TBI后海马神经元凋亡的影响,神经元的免疫荧光染色和Westernblot结果显示：对TBI后的海马神经元给予适当浓度及时程的TGF-β1处理,海马神经元的的凋亡明显降低。结论这些结果的发现,将有助于更好的理解TGF-β1在大鼠创伤性颅脑损伤中的作用,也有望使得TGF—β1成为临床上救治创伤性颅脑损伤的靶点。     

糖皮质激素对重度颅脑损伤患者血浆TNF-α、IL-1β水平的影响

目的 研究糖皮质激素对颅脑损伤患者血浆肿瘤坏死因子α（TNF-α）、白细胞介素-1β（IL-1β）水平的影响。方法 随机将重度颅脑损伤患者分为激素治疗组（20例）与非激素治疗对照组（22例），激素组给予地塞米松10mg／d，共7d。正常组选择健康体检者15例。采用ELSIA法检测两组患者伤后第1、2、7、14天血浆中TNF一仪、IL-1β含量。结果 在颅脑损伤第1、2天激素组血浆TNF-α、IL-1β水平明显高于正常组（P（0．01），在第7、14天明显低于非激素组（P（0．01），但与正常组无明显差异（P〉0．05）。非激素组血浆TNF-α、IL-1β水平在各时间点均明显高于正常组（P〈0．01）。结论 糖皮质激素对降低颅脑损伤患者血浆TNF-α、IL-1β水平具有明显的延迟性，使损伤早期因TNF-α、IL-1β显著升高引起的有害作用未能消除，至恢复期又使TNF-α、IL-1β明显降低，其神经保护作用不能发挥。     

The microglial NLRP3 inflammasome is activated by amyotrophic lateral sclerosis proteins

Microglial NLRP3 inflammasome activation is emerging as a key contributor to neuroinflammation during neurodegeneration. Pathogenic protein aggregates such as β-amyloid and α-synuclein trigger microglial NLRP3 activation, leading to caspase-1 activation and IL-1β secretion. Both caspase-1 and IL-1β contribute to disease progression in the mouse SOD1^G93A model of amyotrophic lateral sclerosis (ALS), suggesting a role for microglial NLRP3. Prior studies, however, suggested SOD1^G93A mice microglia do not express NLRP3, and SOD1^G93A protein generated IL-1β in microglia independent to NLRP3. Here, we demonstrate using Nlrp3-GFP gene knock-in mice that microglia express NLRP3 in SOD1^G93A mice. We show that both aggregated and soluble SOD1^G93A activates inflammasome in primary mouse microglia leading caspase-1 and IL-1β cleavage, ASC speck formation, and the secretion of IL-1β in a dose- and time-dependent manner. Importantly, SOD1^G93A was unable to induce IL-1β secretion from microglia deficient for Nlrp3, or pretreated with the specific NLRP3 inhibitor MCC950, confirming NLRP3 as the key inflammasome complex mediating SOD1-induced microglial IL-1β secretion. Microglial NLRP3 upregulation was also observed in the TDP-43^Q331K ALS mouse model, and TDP-43 wild-type and mutant proteins could also activate microglial inflammasomes in a NLRP3-dependent manner. Mechanistically, we identified the generation of reactive oxygen species and ATP as key events required for SOD1^G93A-mediated NLRP3 activation. Taken together, our data demonstrate that ALS microglia express NLRP3, and that pathological ALS proteins activate the microglial NLRP3 inflammasome. NLRP3 inhibition may therefore be a potential therapeutic approach to arrest microglial neuroinflammation and ALS disease progression.     

SPECT及SOD检测在颅脑损伤患者中的应用

目的探讨超氧化物歧化酶（SOD）及单光子发射计算机体层摄影（SPECT）在评价颅脑损伤后病情和预后中的作用。方法选择符合要求的颅脑损伤患者60例，按照GCS评分标准分成轻型损伤组45例，中型损伤组8例和重型损伤组7例。20例健康员工为对照组。对人选的所有对象进行SPECT、CT扫描和SOD检测，其中颅脑损伤患者在伤后24h内和2周后进行SPECT和CT检查，伤后12h、24h、36h、48h、72h、1周、2周、1月和2-月抽取外周静脉血测定SOD含量。并对所有患者在伤后6月按GOS评价预后。结果入院初和伤后2周，对颅脑损伤患者SPECT检出阳性率要明显高于CT检出阳性率（P〈0．01），尤其对轻型损伤的患者更是如此．SPECT检出的颅脑损伤病灶数也比CT多。外周血SOD含量在伤后24h显著下降（P〈0．05），其中中型和重型损伤组SOD值下降最明显，直到1月后其值才恢复正常；伤后24h外周血SOD值与入院时GCS评分及伤后6月GOS评分呈正相关（P〈0.05）。结论颅脑损伤后早期进行SPECT和SOD的检测可以评价病情等级并可以对预后情况最出初步估计。     

大鼠脑外伤后自噬被激活并在早期对受损神经元起保护作用

目的研究大鼠脑外伤后自噬是否被激活并探讨其在脑外伤后神经细胞损伤和修复中的作用。方法建立大鼠定量脑外伤模型,于脑外伤后不同时间点处死动物并取脑;应用透射电镜检测脑组织自噬双层膜结构以及次级溶酶体的形成情况;应用白噬标记抗体LC3B和DBeclin-1对脑外伤后不同时间点的脑组织进行免疫荧光和Western blot检测;LC3和caspase-3或Beclin1和Fluoro—Jade双标记检测。结果脑外伤后1h在损伤区周围即检测到双层膜结构,并且一直持续到脑外伤后32天。脑外伤后1h,脑组织中LC3和Beclin-1表达增加,损伤后3天内阳性细胞以神经元为主,之后阳性胶质细胞增加,第8天达到高峰,并可持续至脑外伤后32天仍维持高表达。大多数阳性细胞分布在损伤区周围（包括海马）而不是损伤区。此外,脑外伤后24小时以前,在损伤区周围不是所有的LC3阳性细胞都与caspase-3阳性细胞重叠。同样脑外伤后6h至48h,Beclin1阳性海马神经元与Fluoro—Jade染色不重叠。结论脑外伤后自噬被激活,在损伤后早期保护损伤区周围神经细胞免于凋亡和退行性变,并对神经细胞损伤与修复发挥长期作用。     

Transgenic mice that overexpress the anti-apoptotic Bcl-2 protein have improved histological outcome but unchanged behavioral outcome after traumatic brain injury

Increasing evidence suggests that apoptosis is a contributing factor to neuronal cell death in traumatic brain injury (TBI). There is increased expression, cleavage and activation of caspases as well as other proteins known to regulate apoptosis in neurons after TBI. These proteins include the proto-oncogene Bcl-2 which belongs to a family of proteins with both pro- and anti-apoptotic properties. To investigate the role of apoptosis in TBI and the importance of Bcl-2 protein on the severity and outcome of injury, Bcl-2 overexpressing transgenic and wild-type control mice were subjected to the controlled cortical impact model of TBI. There was no significant difference in the cleavage of caspase-3 or caspase-9 detected by Western blotting of hippocampal samples from transgenic or wild-type mice after TBI. Bcl-2 transgenic mice had smaller contusion volumes and increased numbers of surviving neurons in CA2 but not other regions of hippocampus compared to wild-type controls. By contrast, there was no difference in motor function determined by the round beam balance and wire grip tests between transgenic and wild-type mice after TBI. Cognitive function assessed by the Morris water maze was also not different between groups. These results suggest that overexpression of Bcl-2 is only partially neuroprotective and other members of this protein family may prove to be more important in protecting neurons from cell death.     

The protective effect of erdosteine on short-term global brain ischemia/reperfusion injury in rats

Experimental studies have demonstrated that free radicals play a major role on neuronal injury during ischemia/reperfusion (I/R) in rats. Erdosteine is a thioderivative endowed with mucokinetic, mucolytic and free-radical-scavenging properties. The aim of the present study was to investigate the effect of erdosteine treatment against short-term global brain ischemia/reperfusion injury in rats. The study was carried out on Wistar rats divided into four groups. (i) Control group, (ii) ischemia/reperfusion group, (iii) ischemia/reperfusion+erdosteine group, and (iv) erdosteine group. Superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) activities as well as thiobarbituric acid reactive substances (TBARSs) and nitric oxide (NO) levels were analysed in erythrocyte and plasma of rats. Plasma NO levels were significantly higher in the ischemia/reperfusion group than the other groups. The activities of SOD and GSH-Px were decreased, while TBARS levels increased in the ischemia/reperfusion group compared to other groups in both plasma and erythrocyte. The erythrocyte CAT activity was higher in erdosteine group and there was a statistically significant increase, when compared with the erdosteine plus ischemia/reperfusion group. By treating the rats with erdosteine, the depletion of endogenous antioxidant enzymes (SOD, CAT, GSH-Px) and increase of TBARS and NO levels were prevented. This study, therefore, suggests that erdosteine reduces parameters of oxidative stress is well supported by the data.     

大剂量维生素C和维生素E对急性颅脑损伤病人神经损伤、神经营养及氧化应激的影响

目的 探讨大剂量维生素C和维生素E对急性颅脑损伤病人神经损伤、神经营养及氧化应激的影响。方法 2018年1月至2018年11月前瞻性收集84例急性颅脑损伤并随机分为对照组（n=42,接受常规治疗）和观察组（n=42,接受大剂量维生素C和维生素E联合常规治疗）。治疗前、治疗后4、7 d,采用酶联免疫吸附法测定血清神经损伤指标[包括神经元特异性烯醇化酶（NSE）、S100蛋白、脑红蛋白（NGB）、泛素羧基末端水解酶L1（UCH-L1））、神经营养指标[包括神经营养因子-α（NTF-α）、脑源性神经营养因子（BDNF）、神经生长因子（NGF）、胰岛素样生长因子-1（IGF-I）,采用放射免疫沉淀法测定氧化应激指标[包括超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GPx）、过氧化氢酶（CAT）、丙二醛（MDA）、晚期氧化蛋白产物（AOPP）]。结果 治疗后4、7 d,两组血清NSE、S100B、NGB、UCH-L1、MDA、AOPP含量均显著降低（P<0.05）,血清NTF-α、BDNF、NGF、IGF-I、SOD、GPx、CAT含量均显著增高（P<0.05）,而且,观察组均明显优于对照组（P<0.05）。结论 大剂量维生素C和维生素E治疗能够减轻急性颅脑损伤病人神经损伤程度、氧化应激反应并改善神经营养状态。     

Role for mammalian chitinase 3‐like protein 1 in traumatic brain injury

Traumatic brain injury (TBI) is accompanied by inflammatory infiltrates and CNS tissue response. The astrocytosis associated with TBI has been proposed to have both beneficial and detrimental effects on surviving neural tissue. We recently observed prominent astrocytic expression of YKL‐40/chitinase 3‐like protein 1 (CHI3L1) associated with severity of brain injury. The physiological role of CHI3L1 in the CNS is unknown; however, its distribution at the perimeter of contusions and temporal course of expression suggested that in TBI it might be an important component of the astrocytic response to modulate CNS inflammation. To address this hypothesis, we used serially sectioned brains to quantitatively compare the neuropathological outcomes of TBI produced by controlled cortical impact in wild type (WT) and chi3l1 knockout (KO) mice where the murine YKL‐40 homologue, breast regression protein 39 (BRP‐39/CHI3l1), had been homozygously disrupted. At 21 days post‐injury, chi3l1 KO mice displayed greater astrocytosis (increased GFAP staining) in the hemispheres ipsilateral and contralateral to impact compared with WT mice. Similarly, Iba1 expression as a measure of microglial/macrophage response was significantly increased in chi3l1 KO compared with WT in the hemisphere contralateral to impact. We conclude that astrocytic expression of CHI3L1 limits the extent of both astrocytic and microglial/macrophage facets of neuroinflammation and suggests a novel potential therapeutic target for modulating neuroinflammation.     

Simvastatin therapy prevents brain trauma‐induced increases in β‐amyloid peptide levels

Elevations in β‐amyloid peptide (Aβ) levels after traumatic brain injury (TBI) may confer risk for developing Alzheimer's disease in head trauma patients. We investigated the effects of simvastatin, a 3‐hydroxy‐3‐methylglutaryl‐CoA reductase inhibitor, on hippocampal Aβ burden in a clinically relevant head injury/intervention model using mice expressing human Aβ. Simvastatin therapy blunted TBI‐induced increases in Aβ, reduced hippocampal tissue damage and microglial activation, and improved behavioral outcome. The ability of statins to reduce post‐injury Aβ load and ameliorate pathological sequelae of brain injury makes them potentially effective in reducing the risk of developing Alzheimer's disease in TBI patients. Ann Neurol 2009;66:407–414     

脑出血后灶周组织形态和焦亡相关蛋白表达水平的研究

目的 观察自发性脑出血后血肿周围脑组织内神经胶质细胞的组织形态学特点及焦亡相关蛋白caspase-1、IL-18和IL-1β的表达水平变化。方法 高血压性脑出血死亡者(发病后3Symbol～A@33h死亡)的全脑标本5例,出血侧为实验组,非出血侧为对照组; 血肿周围的脑组织(1×1×0.3 cm³)的石蜡切片行常规H&E染色及caspase-1、IL-18和IL-1β蛋白的免疫组化染色,光学显微镜下观察脑组织内神经胶质细胞的组织形态特点及caspase-1、IL-18和IL-1β蛋白的表达水平变化,计数阳性细胞的数量,析因设计的方差分析和费舍尔最小显著差异确定显著性水平。结果 血肿周围脑组织内的神经胶质细胞胀大,核质凝集、碎裂和溶解; caspase-1、IL-18和IL-1β蛋白表达在神经胶质细胞、血管内皮细胞和壁细胞的胞浆内,IL-18和IL-1β蛋白还同时表达在上述细胞的细胞外基质内; caspase-1、IL-18和IL-1β蛋白表达明显增强,阳性细胞显著增多(P Symbol|@@0.01)。结论 自发性ICH后血肿周围脑组织内的神经胶质细胞胀大,核质凝集、碎裂和溶解; caspase-1、IL-18和IL-1β蛋白表达水平显著上调。这提示神经胶质细胞的焦亡途径被启动,其可能是脑出血后脑组织继发性损伤的机制之一。     

右美托咪啶对创伤性脑损伤大鼠模型的神经保护作用实验研究

目的探讨右美托咪啶对创伤性脑损伤组织炎症与细胞凋亡影响。方法建立创伤性脑损伤大鼠模型,设置实验组和对照组,其中实验组予以右美托咪啶6μg/kg腹腔注射干预,对照组予以等量生理盐水腹腔注射,损伤后第3天和第7天,采用ELISA检测损伤组织内肿瘤坏死因子(TNF-α)和白介素-1β(IL-1β)浓度水平,Western-bolt分析损伤组织内凋亡信号关键蛋白caspase-3表达,TUNEL法观察损伤组织细胞凋亡情况。结果右美托咪啶处理可明显降低创伤性脑损伤组织内炎性因子TNF-α和IL-1β浓度(P0.05),减少凋亡信号关键蛋白caspase-3表达(P0.05),显著抑制创伤性脑损伤组织中神经细胞凋亡(P0.05)。结论右美托咪啶处理具有明显神经保护作用,其机制可能与减轻创伤性脑损伤组织炎性反应和抑制神经细胞凋亡有关。     

急性颅脑损伤患者血清及脑脊液中炎性因子的变化及临床意义

目的探讨炎症细胞因子IL-1β,IL-6与颅脑损伤程度的关系及临床意义。方法对近年来我院收治的780例急性颅脑损伤患者按损伤程度分组（轻、中、重及特重型组）,检测血清及脑脊液中IL-1β及IL-6水平,并对比了不同程度急性颅脑损伤患者上述因子水平的差异并分析其临床意义。结果轻型、中型、重型及特重型颅脑损伤患者血清及脑脊液IL-1β及IL-6水平均显著增加,而重型和特重型颅脑损伤患者两种因子水平要显著高于其他患者,死亡患者IL-1β及IL-6水平高于其他所有患者。结论血清及脑脊液中IL-1β及IL-6水平与颅脑损伤的程度密切相关,能够作为临床监测指标。     

葛根素通过Nrf2-ARE信号通路抵抗氧化应激对创伤性脑损伤的神经保护作用

目的 探讨葛根素对创伤性脑损伤(TBI)模型大鼠的神经保护作用及脑组织红系衍生的核因子相关因子2(Nrf2)一抗氧化反应原件(ARE)信号通路参与的机制。方法 选择健康成年雄性大鼠体重250～300 g构建TBI模型,将大鼠分为4组:创伤组(A组)、假手术组(B组)、葛根素治疗的创伤组(C组)和葛根素治疗的假手术组(D组); 通过采用改良的神经功能缺损评分(mNSS)评价神经功能,脑组织干湿重称量法评价脑水肿,Nissl染色,TUNEL染色评价脑损伤体积和神经元的凋亡,使用酶活试剂盒检测损伤48 h后抗氧化酶SOD,GSH,和GSSG的活性以及氧化应激产物MDA和NO的水平,最后使用western blot和RT-PCR的方法检测Nrf2-ARE信号通路及其下游分子HO-1,NQO1的表达水平。结果 TBI手术后损伤组mNSS评分明显增高(P<0.05),葛根素治疗组能够明显降低mNSS评分(P<0.05)。TBI手术后损伤组脑水肿加重及神经元凋亡增加(P<0.05),葛根素治疗组能够明显挽救脑水肿及神经元凋亡(P<0.05)。TBI手术12 h后损伤组脑内抗氧化酶SOD,GSH,和GSSG的活性增加及氧化应激产物MDA和NO的水平升高(P<0.05),葛根素治疗组能够明显降低抗氧化酶SOD,GSH,和GSSG的活性以及氧化应激产物MDA和NO的水平(P<0.05)。western blot和RT-PCR显示葛根素不改变Nrf2的翻译和表达,但是RT-PCR显示葛根素能够明显促进Nrf2-ARE信号通路下游分子HO-1,NQO1的表达。结论 葛根素可能通过Nrf2-ARE信号通路抵抗氧化应激对创伤性脑损伤发挥神经保护作用。     

Impact of pediatric traumatic brain injury on hippocampal neurogenesis

Traumatic brain injury(TBI) is a major cause of mortality and morbidity in the pediatric population. With advances in medical care, the mortality rate of pediatric TBI has declined. However, more children and adolescents are living with TBI-related cognitive and emotional impairments, which negatively affects the quality of their life. Adult hippocampal neurogenesis plays an important role in cognition and mood regulation. Alterations in adult hippocampal neurogenesis are associated with a variety of neurological and neurodegenerative diseases, including TBI. Promoting endogenous hippocampal neurogenesis after TBI merits significant attention. However, TBI affects the function of neural stem/progenitor cells in the dentate gyrus of hippocampus, which results in aberrant migration and impaired dendrite development of adult-born neurons. Therefore, a better understanding of adult hippocampal neurogenesis after TBI can facilitate a more successful neuro-restoration of damage in immature brains. Secondary injuries, such as neuroinflammation and oxidative stress, exert a significant impact on hippocampal neurogenesis. Currently, a variety of therapeutic approaches have been proposed for ameliorating secondary TBI injuries. In this review, we discuss the uniqueness of pediatric TBI, adult hippocampal neurogenesis after pediatric TBI, and current efforts that promote neuroprotection to the developing brains, which can be leveraged to facilitate neuroregeneration.     

Neutralization of interleukin-1β reduces cerebral edema and tissue loss and improves late cognitive outcome following traumatic brain injury in mice

Increasing evidence suggests that interleukin-1β (IL-1β) is a key mediator of the inflammatory response following traumatic brain injury (TBI). Recently, we showed that intracerebroventricular administration of an IL-1β-neutralizing antibody was neuroprotective following TBI in mice. In the present study, an anti-IL-1β antibody or control antibody was administered intraperitoneally following controlled cortical injury (CCI) TBI or sham injury in 105 mice and we extended our histological, immunological and behavioral analysis. First, we demonstrated that the treatment antibody reached target brain regions of brain-injured animals in high concentrations (> 11 nm) remaining up to 8 days post-TBI. At 48 h post-injury, the anti-IL-1β treatment attenuated the TBI-induced hemispheric edema (P < 0.05) but not the memory deficits evaluated using the Morris water maze (MWM). Neutralization of IL-1β did not influence the TBI-induced increases (P < 0.05) in the gene expression of the Ccl3 and Ccr2 chemokines, IL-6 or Gfap. Up to 20 days post-injury, neutralization of IL-1β was associated with improved visuospatial learning in the MWM, reduced loss of hemispheric tissue and attenuation of the microglial activation caused by TBI (P < 0.05). Motor function using the rotarod and cylinder tests was not affected by the anti-IL-1β treatment. Our results suggest an important negative role for IL-1β in TBI. The improved histological and behavioral outcome following anti-IL-1β treatment also implies that further exploration of IL-1β-neutralizing compounds as a treatment option for TBI patients is warranted.     

Underlying mechanism of protection from hypoxic injury seen with n-butanol extract of Potentilla anserine L. in hippocampal neurons

The alcohol and n-butanol extract of Potentilla anserine L. significantly protects myocardium from acute ischemic injury. However, its effects on rat hippocampal neurons and the mechanism of protection remain unclear. In this study, primary cultured hippocampal neurons from neonatal rats were incubated in 95% N2 and 5% CO2 for 4 hours. Results indicated that hypoxic injury decreased the viability of neurons, increased the expression levels of caspase-9 and caspase-3 mRNA, as well as cytochrome c, Caspase-9, and Caspase-3 protein. Pretreatment with 0.25, 0.062 5, 0.015 6 mg/mL n-butanol extract of Potentilla anserine L. led to a significant increase in cell viability. Expression levels of caspase-9 and caspase-3 mRNA, as well as cytochrome c, Caspase-9, and Caspase-3 protein, were attenuated. The neuroprotective effect of n-butanol extract of Potentilla anserine L. was equivalent to tanshinone IIA. Our data suggest that the n-butanol extract of Potentilla anserine L. could protect primary hippocampal neurons from hypoxic injury by deactivating mitochondrial cell death.     

Altered antioxidant defense system in clinically stable patients with schizophrenia and their unaffected siblings

Objective

To determine Red Blood Cell (RBC) antioxidant enzyme activities and plasma Thiobarbituric Acid Reactive Substances (TBARS) in clinically stable patients with schizophrenia and their unaffected siblings.

Methods

A case-control study carried out on three groups: 60 schizophrenic patients treated with neuroleptics, 33 of their unaffected siblings and 30 healthy controls with no family psychiatric history. Biological markers were measured on fasting patients after a period of tobacco abstinence: RBC antioxidant enzyme activities – superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT) – by spectrophotometry and plasma levels of TBARS by spectrofluorimetry.

Results

RBC SOD and CAT activities were significantly lower in schizophrenic patients and their unaffected siblings compared to the control group (P < 0.001). Schizophrenic patients also had significantly lower RBC GSH-Px activity than controls (P = 0.03), whereas their unaffected siblings had significantly higher RBC GSH-Px activity than controls (P = 0.04). Plasma TBARS were higher in schizophrenic patients than their unaffected siblings: 2.1 ± 0.8 μmol/l vs. 1.7 ± 0.6 μmol/l (P = 0.06).

Conclusions

Our results showed a decrease in antioxidant enzyme activities and an increase in lipid peroxidation confirming the existence of oxidative stress in schizophrenic patients treated with neuroleptics. Additionally, this suggests that the increase in GSH-Px activity in unaffected siblings would be a protective mechanism against oxidative stress and damage. Other studies are necessary to confirm these findings.     

Inflammatory cytokine receptor blockade in a rodent model of mild traumatic brain injury

In rodent models of traumatic brain injury (TBI), both Interleukin‐1β (IL‐1β) and tumor necrosis factor‐α (TNFα) levels increase early after injury to return later to basal levels. We have developed and characterized a rat mild fluid percussion model of TBI (mLFP injury) that results in righting reflex response times (RRRTs) that are less than those characteristic of moderate to severe LFP injury and yet increase IL‐1α/β and TNFα levels. Here we report that blockade of IL‐1α/β and TNFα binding to IL‐1R and TNFR1, respectively, reduced neuropathology in parietal cortex, hippocampus, and thalamus and improved outcome. IL‐1β binding to the type I IL‐1 receptor (IL‐1R1) can be blocked by a recombinant form of the endogenous IL‐1R antagonist IL‐1Ra (Kineret). TNFα binding to the TNF receptor (TNFR) can be blocked by the recombinant fusion protein etanercept, made up of a TNFR2 peptide fused to an Fc portion of human IgG1. There was no benefit from the combined blockades compared with individual blockades or after repeated treatments for 11 days after injury compared with one treatment at 1 hr after injury, when measured at 6 hr or 18 days, based on changes in neuropathology. There was also no further enhancement of blockade benefits after 18 days. Given that both Kineret and etanercept given singly or in combination showed similar beneficial effects and that TNFα also has a gliotransmitter role regulating AMPA receptor traffic, thus confounding effects of a TNFα blockade, we chose to focus on a single treatment with Kineret. © 2015 Wiley Periodicals, Inc.