丹酚酸B对小鼠创伤性脑损伤的保护作用

目的通过小鼠控制性皮层撞击(CCI)模型研究丹酚酸B(SalB)对创伤性脑损伤(TBI)的保护作用。方法采用脑含水量测定、运动功能评分、水迷宫测试评价丹酚酸B对小鼠TBI的保护作用。采用酶联免疫吸附法(ELISA)检测炎症因子肿瘤坏死因子α(TNF-α)和白介素1β(IL-1β)的表达水平,评价丹酚酸B对TBI后脑组织炎症反应的抑制作用。结果丹酚酸B显著减轻TBI后脑水肿和运动功能缺损,改善学习记忆功能,并抑制炎症因子TNF-α和IL-1β的表达。结论丹酚酸B对小鼠TBI具有保护作用,这种保护作用可能与其抗炎作用相关。     

甘氨酸通过抑制大鼠创伤性脑损伤后炎症反应来发挥神经保护作用

目的 探讨甘氨酸对创伤性脑损伤(TBI)大鼠的神经保护作用及机制。方法 将SD雄性大鼠随机分为对照(Sham)组、脑损伤+溶剂(TBI+Vehicle)组和脑损伤+甘氨酸(TBI+Glycine)组,采用Feeney's自由落体法建立创伤性脑损伤模型; 术后1 h侧脑室注射甘氨酸(2 mg/kg)或等体积的溶剂; 术后24 h,取脑组织样本; 采用脑含水量测定、蛋白免疫印迹法和免疫荧光法评价甘氨酸对大鼠TBI的神经保护作用; 采用ELISA法检测炎症因子白介素1β(IL-1β)、白介素6(IL-6)和肿瘤坏死因子α(TNF-α)的表达水平,评价甘氨酸对TBI后脑组织炎症反应的抑制作用。结果 甘氨酸可减轻TBI后脑水肿,减少皮层神经元损伤; 同时甘氨酸可抑制TBI后炎症因子IL-1β、IL-6和TNF-α的过度释放。结论 甘氨酸对大鼠TBI具有神经保护作用; 甘氨酸对TBI后相关炎症因子过度增高的抑制可能部分解释其神经保护作用机制。     

姜黄素对APP/PS1双转基因小鼠认知功能、炎症反应及海马区突触素表达的影响

目的 探讨姜黄素对APP/PS1双转基因小鼠认知功能、炎症反应及海马区突触素表达的影响。方法 60只6月龄APP/PS1双转基因雄性小鼠随机分为A组、B组和C组,各20只;A组采用姜黄素100mg/kg/d加入小鼠饲料喂养,B组采用姜黄素300mg/kg/d喂养,C组采用姜黄素600mg/kg/d喂养;3组均喂养6个月。治疗前、治疗3、6个月,采用Morris水迷宫实验评估小鼠认知功能,采用免疫吸附试验法检测尾静脉血血清白介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）等炎症因子水平;治疗6个月,采用免疫组化染色法检测小鼠海马 CA1 区突触素表达情况。结果 姜黄素治疗3、6个月,3组小鼠认知功能明显改善（P<0.05）,血清IL-6、TNF-α水平明显降低（P<0.05）。与A组比较,B组和C组治疗3、6个月认知功能明显改善（P<0.05）,血清IL-6、TNF-α水平均明显降低（P<0.05）,海马CA1区突触素表达水平明显升高（P<0.05）。而B组与C组均无统计学差异（P>0.05）。结论 姜黄素有助于改善APP/PS1双转基因小鼠海马突触素表达,抑制小鼠炎症反应,改善小鼠认知功能。     

TLR4基因突变对小鼠坐骨神经损伤修复的影响

目的 探讨Toll样受体4（TLR4）基因突变达对小鼠坐骨神经损伤修复的影响。方法 取10只C3H/HeJ小鼠（TLR4基因突变作为突变组,20只C3H/HeN小鼠（TLR4基因正常）随机分为假手术组（n=10）和模型组（n=10）。突变组和模型组在暴露的坐骨神经中部用止血钳夹持60 s以建立小鼠坐骨神经损伤模型,假手术组仅暴露坐骨神经而不进行夹伤。造模后4周,采用坐骨神经功能指数（SFI）评分评定坐骨神经功能,然后每组取3只小鼠行HE染色观察坐骨神经病理改变,每组取3只小鼠采用RT-PCR检测坐骨神经组织白细胞介素-1β（IL-1β）、肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）mRNA表达水平,每组取4只小鼠采用免疫印迹法检测坐骨神经组织生长相关蛋白43（GAP43）、p75神经营养素受体（p75NTR）蛋白表达水平。结果 与假手术组相比,模型组SFI评分显著降低（P<0.05）,HE染色显示细胞形态异常并出现大量嗜中性粒细胞和巨噬细胞,坐骨神经组织IL-1β、IL-6、TNF-α mRNA表达水平以及GAP43、p75NTR蛋白表达水平均显著增高（P<0.05）。与模型组相比,突变组SFI评分明显增高（P<0.05）,组织结构病理改变明显改善,IL-1β、IL-6、TNF-α mRNA表达水平以及GAP43、p75NTR蛋白表达水平显著降低（P<0.05）。结论 TLR4基因突变可促进小鼠坐骨神经损伤后修复,可能与降低IL-1β、IL-6、TNF-α等炎症因子水平有关。     

α-亚麻酸对小鼠创伤性脑损伤后急性炎症及神经功能的影响

目的探讨α-亚麻酸(α-linolenic acid,ALA)对小鼠创伤性脑损伤(traumatic brain injury,TBI)后急性炎症及神经功能的影响。方法将C57BL6/N小鼠分为高ALA饮食组(孕鼠20只,新生鼠88只)和低ALA饮食组(孕鼠20只,新生鼠84只),采用气相色谱法检测两组小鼠脑多不饱和脂肪酸含量;运用控制性皮质撞击建立小鼠TBI模型。采用实时定量聚合酶链反应、酶联免疫吸附法和蛋白质印迹法分别检测TBI后0、4、24、96h两组小鼠炎症因子及细胞标志物表达。运用转轮实验、横木行走实验和场景恐惧实验观察两组小鼠TBI后神经功能恢复情况。结果高ALA饮食组小鼠脑二十二碳六烯酸(docosahexaenoic acid,DHA)含量高于低ALA饮食组[15.48%±1.20%、9.98%±1.10%,P0.05];高ALA饮食组小鼠TBI后TNF-α和IL-1β、IL-6和CCL12显著低于低ALA饮食组(P0.05)。高ALA饮食组小鼠TBI 24h后运动功能恢复较低ALA饮食组快。高ALA饮食组小鼠TBI 24 h后认知功能均优于低ALA饮食组。结论增加脑部DHA水平可以减少急性炎症和改善TBI后神经功能恢复。     

葛根素对大鼠颅脑损伤的保护作用

目的 探讨葛根素对大鼠颅脑损伤（TBI）的保护作用及其机制。方法 将45只SD大鼠随机分为假手术组、模型组、低剂量葛根素组、中剂量葛根素组和高剂量葛根素组,每组9只。采用Feeney氏自由落体法制备TBI大鼠模型。低、中、高剂量葛根素组腹腔注射葛根素,剂量分别为10、25、50 mg/kg。造模后1、3、7 d采用改良神经功能缺损评分（mNSS）评价神经功能。造模后7 d,干湿法测定脑组织含水量;ELISA法检测脑组织丙二醛（MDA）、超氧化物歧化酶（SOD）、谷胱甘肽（GSH）、过氧化氢酶（CAT）、核因子κB （NF-κB）、细胞间黏附分子-1（ICAM-1）、白介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）、caspase-3水平;免疫印迹法检测Bax、Bcl-2的表达。结果 葛根素能显著降低TBI大鼠mNSS（P<0.05）,显著减轻脑组织水肿（P<0.05）,显著降低脑组织MDA、SOD、GSH、CAT、NF-κB、ICAM-1、IL-6、TNF-α、caspase-3水平（P<0.05）,显著下调Bax表达而上调Bcl-2表达（P<0.05）。结论 葛根素可通过减轻颅脑水肿、抑制氧化应激及炎性反应以及调节Bax/Bcl-2表达从而发挥神经保护作用。     

丹酚酸B对活化血小板诱导脑微血管内皮细胞炎症应答的影响

目的 研究丹酚酸B对活化血小板诱导的人脑微血管内皮细胞(HBMECs)炎症应答的干预效应.方法 体外激活的血小板与HBMECs共培养12h,建立炎症应答模型,实时荧光定量RT-PCR检测HBMECs中细胞间黏附分子-1(ICAM-1)、IL-1β、Ⅱ-6以及Ⅱ-8、趋化蛋白-I(MCP-1)mRNA表达水平.同时采用10 μg/mL丹酚酸B预适应HBMECs 24h,检测上述炎症介质表达水平的变化.结果 丹酚酸B预适应可不同程度减少活化血小板诱导的HBMECs炎症模型中ICAM-1、IL-1β、IL-6、IL-8、MCP-1 mRNA表达水平,与干预前比较,差异均有统计学意义(P＜0.05).结论 丹酚酸B可阻抑活化血小板诱导的脑微血管内皮细胞炎症应答,推测是其治疗脑缺血的机制之一.     

美满霉素抑制血管性认知功能损伤大鼠海马星型胶质细胞激活和神经炎症(英文)

目的观察美满霉素(minocycline)对血管性认知功能损伤大鼠海马组织GFAP、COX-2、NF-κB、IL-1β和TNF-α表达的影响,探讨美满霉素对血管性认知功能损伤脑保护作用的机制。方法Wistar大鼠随机分为假手术组(S组)、血管性认知功能损伤模型组(M组)和美满霉素治疗组(MT组)。免疫组织化学法检测大鼠海马组织COX-2和NF-κB的表达,蛋白质印迹和免疫组织化学法检测大鼠海马组织GFAP的表达,ELISA法检测大鼠海马组织IL-1β和TNF-α的表达。结果MT 组 GFAP、COX-2、NF-κB、IL-1β和 TNF-α表达较 M 组均降低(P<0.01) ;MT 和 M 组GFAP、COX-2、NF-κB、IL-1β和 TNF-α表达均显著高于 S 组(P<0.01)。结论美满霉素能降低血管性认知功能损伤大鼠海马组织中GFAP、COX-2、NF-κB、IL-1β和TNF-α的表达,抑制血管性认知功能损伤大鼠海马星型胶质细胞活化和神经炎症,发挥脑保护作用。     

阿司匹林对脑缺血炎症反应的抑制作用

目的探讨阿司匹林对脑缺血再灌注损伤炎症反应的影响及其机制.方法采用线栓法制备短暂性大脑中动脉缺血模型,用免疫组织化学染色观察脑组织中核转录因子-κB(NF-κB)活性、白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)的表达;HE染色观察中性粒细胞浸润.结果与对照组比较,小剂量阿司匹林组和大剂量阿司匹林组的NF-κB活性降低,以大剂量的阿司匹林作用更为明显;各阿司匹林组的IL-1β、TNF-α的表达明显降低,白细胞浸润明显减少.结论阿司匹林可抑制脑缺血再灌注损伤过程中的炎症反应;其机制可能与抑制NF-κB的激活和IL-1β、TNF-α表达有关.     

黄芩苷对海人酸致痫小鼠海马白细胞介素-1β及肿瘤坏死因子-α表达的影响

目的 探讨黄芩苷对海人酸诱导的小鼠癫痫持续状态后海马组织白细胞介素-1 β(IL-1β)、肿瘤坏死因子-α(TNF-α)表达的影响.方法 将54只ICR雄性小鼠随机分为对照组、癫痫持续状态(SE)组、黄芩苷治疗组,每组18只.采用侧脑室注入海人酸建立小鼠癫痫持续状态模型.HE染色观察黄芩苷对小鼠癫痫持续状态后海马神经细胞的形态学影响.通过RT-PCR和Western blot分别检测小鼠海马组织中IL-1β mRNA、TNF-α mRNA及IL-1β、TNF-α蛋白的表达量.结果 黄芩苷明显改善了SE后小鼠海马组织的病理形态学,并且可降低IL-1β、TNF-α的表达(P＜0.05).结论 黄芩苷可能通过降低癫痫鼠海马组织中IL-1 β、TNF-α的表达发挥抗炎作用,从而对脑组织进行保护.     

Salvianolic acid B attenuates brain damage and inflammation after traumatic brain injury in mice

Salvianolic acid B (SalB), a bioactive compound isolated from the Chinese medicinal herb Danshen, has been shown to exert various anti-oxidative and anti-inflammatory activities in in vitro and in vivo studies. Here, we investigated the protective effects of SalB on traumatic brain injury (TBI) in mice. When administered within 2 h after TBI onset, SalB (25 mg/kg) reduced brain edema, lesion volume and motor functional deficits, and improved spatial learning and memory abilities. Moreover, SalB treatment inhibited the neutrophil infiltration and microglial activation at 48 h after TBI. Enzyme-linked immunosorbent assay (ELISA) for brain tissue homogenates was performed at 24 h after TBI to evaluate the expression of inflammation-related cytokines. The results showed that SalB suppressed the expression of pro-inflammatory cytokines TNF-α and IL-1β, whereas enhanced the expression of anti-inflammatory cytokines IL-10 and TGF-β1. All of these findings extended the protective role of SalB in the model of TBI and suggested that these protective effects might be associated with its anti-inflammatory activities. Thus SalB may have therapeutic potential for patients with TBI and perhaps other forms of acute brain injury.     

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。     

偏执型分裂症外周血IL-1β、TNF-α和酪氨酸羟化酶mRNA表达水平与临床症状的关系

目的检测偏执型精神分裂症患者白介素-1β、肿瘤坏死因子-α和酪氨酸羟化酶（TH）的基因表达水平,探讨其与临床症状的关系。方法采用RT-PCR和半定量技术,分别检测39例偏执型精神分裂症患者和30例正常对照外周血单个核细胞IL-1β、TNF-α和TH的基因表达水平,同时应用PANSS量表评定偏执型精神分裂症患者临床症状。结果研究显示病例组的IL-1β、TNF-α、TH基因表达水平均显著高于正常对照组（P〈0.01）。且同时发现IL-1β（r=0.420）、TNF-α（r=0.430）基因表达水平与PANSS量表的一般病理症状分显著相关（P〈0.01）。结论偏执型精神分裂症患者可能存在致炎性细胞因子和儿茶酚胺类神经递质的过度表达;致炎性细胞因子可能参与偏执型精神分裂症一般病理症状的形成。     

高血压脑出血患者血清和颅内血肿液中IL-1β、IL-6、TNF—α的含量研究

目的通过测定高血压脑出血患者静脉血清及颅内血肿液中白细胞介素-1β（IL-1β）、白细胞介素．6（IL-6）、肿瘤坏死因子-α（TNF—α）水平,探讨这些炎症因子与脑出血病灶及外周血之间的关系。方法在高血压脑出血后24h,72h,7d和14d取80例高血压脑出血患者静脉血清及血肿液检测IL-1β、IL-6、TNF-α的含量（实验组）,并与30例正常体检的患者（对照组）对比。结果高血压脑出血患者血肿液IL-1β、IL-6、TNF-α含量显著高于对照组人群静脉血,并且显著高于实验组患者自身静脉血。实验组患者静脉血清中IL-1β、IL-6、TNF-α含量显著高于对照组人群静脉血清,且病情越重几种炎症因子水平越高。脑出血不同时期静脉血清IL-1β、IL-6、TNF—α．含量比较,在72h最高,此后呈下降趋势。结论高血压脑出血患者IL-1β、IL-6、TNF-α参与了脑出血的炎症反应,是脑出血后脑组织损伤的重要机制之一。     

Postinjury treatment with rolipram increases hemorrhage after traumatic brain injury

The pathology caused by traumatic brain injury (TBI) is exacerbated by the inflammatory response of the injured brain. Two proinflammatory cytokines that contribute to inflammation after TBI are tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β). From previous studies using the parasagittal fluid-percussion brain injury model, we reported that the anti-inflammatory drug rolipram, a phosphodiesterase 4 inhibitor, reduced TNF-α and IL-1β levels and improved histopathological outcome when administered 30 min prior to injury. We now report that treatment with (±)-rolipram given 30 min after injury significantly reduced TNF-α levels in the cortex and hippocampus. However, postinjury administration of (±)-rolipram significantly increased cortical contusion volume and increased atrophy of the cortex compared with vehicle-treated animals at 10 days postinjury. Thus, despite the reduction in proinflammatory cytokine levels, histopathological outcome was worsened with post-TBI (±)-rolipram treatment. Further histological analysis of (±)-rolipram-treated TBI animals revealed significant hemorrhage in the contused brain. Given the well-known role of (±)-rolipram of increasing vasodilation, it is likely that (±)-rolipram worsened outcome after fluid-percussion brain injury by causing increased bleeding.     

Exercise Pre-conditioning Reduces Brain Inflammation and Protects against Toxicity Induced by Traumatic Brain Injury: Behavioral and Neurochemical Approach

Although the favorable effects of physical exercise in neurorehabilitation after traumatic brain injury (TBI) are well known, detailed pathologic and functional alterations exerted by previous physical exercise on post-traumatic cerebral inflammation have been limited. In the present study, it is showed that fluid percussion brain injury (FPI) induced motor function impairment, followed by increased plasma fluorescein extravasation and cerebral inflammation characterized by interleukin-1β, tumor necrosis factor-α (TNF-α) increase, and decreased IL-10. In addition, myeloperoxidase (MPO) increase and Na?,K?-ATPase activity inhibition after FPI suggest that the opening of blood-brain barrier (BBB) followed by neurtrophils infiltration and cerebral inflammation may contribute to the failure of selected targets leading to secondary damage. In fact, Pearson's correlation analysis revealed strong correlation of MPO activity increase with Na?,K?-ATPase activity inhibition in sedentary rats. Statistical analysis also revealed that previous running exercise (4 weeks) protected against FPI-induced motor function impairment and fluorescein extravasation. Previous physical training also induced IL-10 increase per se and protected against cerebral IL-1β, and TNF-α increase and IL-10 decrease induced by FPI. This protocol of physical training was effective against MPO activity increase and Na?,K?-ATPase activity inhibition after FPI. The present protection correlated with MPO activity decrease suggests that the alteration of cerebral inflammatory status profile elicited by previous physical training reduces initial damage and limits long-term secondary degeneration after TBI. This prophylactic effect may facilitate functional recovery in patients suffering from brain injury induced by TBI.     

几种激素和IL-1β对人脑星形胶质细胞IL-6 、TNF-α分泌的影响

目的 观察T3等因素对体外培养人胎大脑星形胶质细胞分泌IL-6、TNF-α的调节作用。方法 纯化培养人胎大脑星形胶质细胞,应用酶联免疫分析(ELISA)方法检测培养上清液中IL-6、TNF-α的水平。结果 (1)星形胶质细胞(AC)在体外培养条件下可自发分泌IL-6,而TNF-α则几乎检测不到。(2)LPS(0.1μg/mL)即可诱导AC产生IL-6和TNF-α。(3)IL-1β是IL-6分泌的主要诱导剂,但不诱导TNF-α分泌。(4)氢化可的松可明显抑制AC分泌IL-6、TNF-α。(5)T3在72h可刺激IL-6的分泌。(6)胰岛素对IL-6的分泌没有明显的调节作用。结论 AC可通过分泌细胞因子参与炎症反应等病理过程并维持中枢神经系统的正常发育、内环境的稳定,且受多种因素的调节。在中枢神经系统中T3、胰岛素主要参与调节发育和代谢,可能不直接参与炎症和免疫机制调节。     

Propofol administration improves neurological function associated with inhibition of pro-inflammatory cytokines in adult rats after traumatic brain injury

BackgroundNeurological deficits following traumatic brain injury (TBI) result in dramatic impacts on the survivors, but the effect of propofol and associated mechanism are waiting to be determined.MethodsAdult male Sprague–Dawley rats were randomly assigned into Sham, TBI, TBI + Intralipid and TBI + Propofol group. Modified Feeney method was adopted to generate TBI model from free hammer fall injury, and animals in TBI + Propofol group were immediately treated with propofol administration for 2 hours after TBI, rats after TBI without propofol treatment was used as injury control, intralipid as vehicle in propofol was injected in TBI + intralipid group. Then, neurological severity scores (NSS) were evaluated at 1, 3, 7 and 14 days. Moreover, the expressions of IL-1β, IL-6 and TNF-α mRNA and protein were examined using quantitative real time-polymerase chain reaction and Western blot, immunohistochemical staining was used to localize cytokines.ResultsThe NSS increased greatly in the rats induced by TBI, while propofol could effectively decreased NSS, confirming the neuroprotective effect of propofol. Moreover, the mRNA expressions of IL-1β, IL-6 and TNF-α, at 1, 3, 7 days after operation (dpo), were significantly augmented in the injured cortex, compared with sham one. But there was no difference between TBI and TBI + Intralipid group, but markedly decreased after propofol treatment. Additionally, the protein level of IL-1β, IL-6 and TNF-α in four groups determined by Western blot and immunohistochemistry showed the similar change with mRNA expression.ConclusionPropofol treatment could elicit a robust neuroprotective response, resulting in significant neurological function improvement for TBI rats, which was independent with intralipid. The underlying molecular mechanism may be partially associated with an inhibition of pro-inflammatory cytokines.