JNK-IN-8, a c-Jun N-terminal kinase inhibitor,improves functional recovery through suppressing neuroinflammation in ischemic stroke

C-Jun N-terminal kinase (JNK) is a pivotal MAPK (mitogen-activated protein kinase), which activated by ischemia brain injury and plays a fairly crucial function in cerebral ischemic injury. Emerging studies demonstrated that JNK-IN-8 (a JNK inhibitor with high specificity) regulates traumatic brain injury through controlling neuronal apoptosis and inflammation. However, the function of JNK-IN-8 in ischemic stroke and the mechanisms underlying of JNK-IN-8 about neuroprotection are not well understood. In this work, male rats were treated with JNK-IN-8 after transient middle cerebral artery occlusion, and then the modified improved neurological function score (mNSS), the foot-fault test (FFT), interleukin-1β (IL-1β), IL-6, and tumor necrosis factor-α (TNF-α) levels were assessed. We found that JNK-IN-8-treated rats with MCAO exerted an observable melioration in space learning as tested by the improved mNSS, and showed sensorimotor functional recovery as measured by the FFT. JNK-IN-8 also played anti-inflammatory roles as indicated through decreased activation of microglia and decreased IL-6, IL-1β, and TNF-α expression. Furthermore, JNK-IN-8 suppressed the activation of JNK and nuclear factor-κB (NF-κB) signaling as indicated by the decreased level of phosphorylated-JNK and p65. All data demonstrate that JNK-IN-8 inhibits neuroinflammation and improved neurological function by inhibiting JNK/NF-κB and is a promising agent for the prevention of ischemic brain injury.     

蜂胶黄酮对大鼠脑缺血再灌注损伤后IL-1β、IL-6和TNF-α含量的影响

目的探讨蜂胶黄酮对脑缺血再灌注损伤的保护作用。方法采用大鼠大脑中动脉栓塞模型（MCAO）,研究蜂胶黄酮对脑缺血再灌注脑梗死体积和行为学评分的影响,对脑组织内IL-1β、IL-6和TNF-α含量的影响。结果蜂胶黄酮能够减少MCAO脑梗死体积和行为学评分,降低脑组织中IL-1β、IL-6和TNF-α的含量（P〈0．05）。结论蜂胶黄酮对大鼠脑缺血再灌注损伤有一定保护作用,其作用机制与降低脑组织中IL-1β、IL-6和TNF-α含量有关。     

L-Arg对LPS诱导的急性肺损伤大鼠肺表面活性物质和肺泡巨噬细胞功能的影响

目的:探讨L-精氨酸(L-Arg)对脂多糖(LPS)诱导的急性肺损伤大鼠肺表面活性物质和肺泡巨噬细胞功能的影响。方法:舌下静脉注射脂多糖(LPS)复制肺损伤模型。健康雄性SD大鼠48只,随机分为对照组、模型组(LPS组)和L-Arg治疗组(L-Arg组)(n=16)。分别于给予LPS 3 h或6 h后给予生理盐水(对照组及LPS组,ip)和L-Arg(500 mg/kg ip)(L-Arg治疗组),治疗3 h。原位杂交法(ISH)检测肺组织中肺表面活性蛋白A(SP-A)mRNA的表达;测定肺泡灌洗液(BALF)中的总蛋白(TP)。体外分离培养大鼠肺泡巨噬细胞,以LPS(终浓度10 mg/L)处理巨噬细胞,观察L-Arg对肺泡巨噬细胞的影响。结果:与对照组比较,大鼠肺损伤后SP-A mRNA表达减弱,BALF中TP增多(P<0.01)。肺损伤3 h用L-Arg治疗3 h后,SP-A mRNA阳性细胞表达明显增强,BALF中TP较LPS组相同时间点明显降低(P<0.05,P<0.01),肺损伤减轻。体外实验中,与正常对照组相比,LPS组细胞培养上清中乳酸脱氢酶(LDH)、一氧化氮(NO)、肿瘤坏死因子-α(TNFα-)和白细胞介素-6(IL-6)浓度明显增高(P<0.01);L-Arg明显减少LPS所致的LDH的释放,降低TNFα-和IL-6浓度。结论:L-Arg可减轻内毒素性肺损伤,此机制可能与增强SP-AmRNA表达有关;LPS可刺激巨噬细胞分泌促炎因子和NO,L-Arg可抑制LPS对巨噬细胞的作用。     

PM2.5鼻腔滴注致小鼠海马组织损伤的炎性机制

目的: 探讨鼻腔滴注不同浓度的PM_2.5对小鼠海马组织损伤的炎性机制。方法: 30只C57BL/6J小鼠随机分为3组(n=10):对照组、低剂量组、高剂量组。采用鼻腔滴注方法进行染毒,每次染毒前测量体重,低剂量组和高剂量组PM_2.5染毒剂量分别为1.5 mg/kg BW和7.5 mg/kg BW,对照组给予等体积的生理盐水,隔天染毒一次,共12次。用ELISA法测定血清中肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)和白介素-6(IL-6)水平。HE染色和电镜观察肺和海马组织的病理变化及超微结构。用抗体芯片技术测定海马组织炎性细胞因子水平。结果: PM_2.5鼻腔滴注染毒对小鼠血清中TNF-α、IL-1β和IL-6水平无显著影响(P＞0.05),肺组织结构也无明显病理改变。而在海马组织中,低剂量和高剂量PM_2.5暴露均能导致海马CA3区神经元排列紊乱,并存在神经元细胞周围突触数量减少,小血管周围水肿等超微结构变化。利用抗体芯片检测海马组织炎性细胞因子表达变化,结果显示,与对照组比较,低剂量组海马组织中CX3CL1、CSF2和TECK等炎性细胞因子水平显著升高(P＜0.05),而MIG和sTNFR1显著降低(P＜0.05);高剂量组中炎性因子CX3CL1、CSF2和TCA-3等显著升高(P＜0.05),而Leptin、MIG和FASLG等显著降低(P＜0.05)。结论: PM_2.5鼻腔滴注可诱导小鼠海马组织结构损伤,其作用途径可能为嗅脑通路,引起海马损伤的炎性机制可能与TNF-α和IL-6等促炎因子的显著升高和sTNFR1 、FASLG等炎性疾病标志物的显著降低有关。     

芪蝎活血通络汤对局灶性脑缺血再灌注损伤模型大鼠神经功能、氧化应激及炎症因子的影响

目的:探讨芪蝎活血通络汤对局灶性脑缺血再灌注损伤模型大鼠神经功能、氧化应激及炎症因子的影响。方法:50只SD大鼠随机选取40只采用线栓法构建脑缺血再灌注模型,其中36只成功,4只死亡。随机数字表法将36只脑缺血再灌注模型大鼠分为模型组、低剂量组(芪蝎活血通络汤2.0 mg/kg灌胃处理)、中剂量组(芪蝎活血通络汤4.0 mg/kg灌胃处理)和高剂量组(芪蝎活血通络汤8.0 mg/kg灌胃处理),每组9只,剩余10只大鼠仅切开皮肤分离和夹闭血管(对照组)。建模后芪蝎活血通络汤各剂量组给予相应剂量灌胃,对照组和模型组给予同剂量生理盐水灌胃,持续4周。用药4周后测评各组大鼠神经功能缺损程度、双侧贴纸去除时间、平衡木过杆时间,并测定各组大鼠脑组织中含水量、脑梗死面积以及氧化应激[过氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、过氧化氢酶(CAT)、丙二醛(MDA)]和炎症因子[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)]指标。结果:与对照组比较,模型组大鼠m NSS评分、脑组织含水量、MDA含量、IL-6、IL-1β、TNF-α水平升高(P<0.05),双侧贴纸去除时间、平衡木过杆时间延长(P<0.05),脑梗死面积增大(P<0.05),SOD、GSH-Px、CAT活性降低(P<0.05);与模型组比较,芪蝎活血通络汤各剂量组大鼠m NSS评分、脑组织含水量、MDA含量、IL-6、IL-1β、TNF-α水平降低(P<0.05),双侧贴纸去除时间、平衡木过杆时间缩短(P<0.05),脑梗死面积缩小(P<0.05),SOD、GSH-Px、CAT活性升高(P<0.05);与低剂量组比较,中、高剂量组大鼠m NSS评分降低(P<0.05),双侧贴纸去除时间、平衡木过杆时间缩短(P<0.05);高剂量组大鼠脑梗死面积、脑组织MDA、IL-6、IL-1β、TNF-α低于低剂量组(P<0.05),SOD、GSH-Px、CAT高于低剂量组(P<0.05)。结论:芪蝎活血通络汤可降低大鼠脑缺血再灌注损伤,改善神经功能,其治疗作用可能与抗氧化,抗炎作用有关,8.0 mg/kg剂量效果最显著。     

丙泊酚对全肝缺血再灌注大鼠脑损伤的保护作用及机制研究

目的:探究丙泊酚对全肝缺血再灌注(THIR)大鼠脑损伤的保护作用及机制。方法:选取72只健康成年雄性SD大鼠,将其按照抽签法分成假手术组、对照组以及丙泊酚组。所有大鼠予以12h禁食处理,采用3%戊巴比妥钠行腹腔注射麻醉处理,常规消毒后取上腹部正中切口进入腹腔。假手术组仅暴露肝门,不予以阻断处理。对照组与丙泊酚组则以无创动脉夹阻断肝固有动脉、门静脉和胆总管,在右肾动脉水平处阻断肝下下腔静脉,膈肌水平阻断肝上下腔静脉,进入全肝缺血阶段,阻断30 min后去除动脉夹恢复肝血流。其中丙泊酚组在全肝缺血前10 min予以丙泊酚50 mg/kg腹腔注射干预,假手术组与对照组则予以等量的生理盐水腹腔注射干预。比较三组大鼠再灌注24h后的脑组织细胞凋亡率、特异性半胱氨酸蛋白酶-3(Caspase-3)蛋白表达水平,脑组织超氧化物歧化酶(SOD)、丙二醛(MDA)、一氧化氮(NO)水平,血清白介素-6(IL-6)以及肿瘤坏死因子-α(TNF-α)水平。结果:对照组与丙泊酚组大鼠的细胞凋亡率及Caspase-3相对表达量均高于假手术组,而丙泊酚组细胞凋亡率及Caspase-3相对表达量均低于对照组(均P<0.05)。对照组与丙泊酚组大鼠脑组织SOD水平均低于假手术组,而丙泊酚组脑组织SOD水平高于对照组;对照组与丙泊酚组大鼠脑组织MDA、NO水平均高于假手术组,而丙泊酚组脑组织MDA、NO水平低于对照组(均P<0.05)。对照组与丙泊酚组大鼠血清IL-6、TNF-α水平均高于假手术组,而丙泊酚组血清IL-6、TNF-α水平均低于对照组(均P<0.05)。结论:丙泊酚可有效抑制THIR大鼠脑损伤引起的细胞凋亡,其主要机制可能与抑制Caspase-3表达、炎症反应以及抗自由基损伤有关。     

急性颅脑损伤大鼠脑组织ATP酶活性及TNF-α的变化及意义

目的 观察大鼠急性颅脑损伤后脑组织ATP酶活性及肿瘤坏死因子-α（TNF-α）的变化,探讨急性颅脑损伤后脑水肿的发病机制.方法 将72只SD大鼠随机分为正常组（N组）、假手术对照组（S组）、急性颅脑损伤模型组（ACI组）,其中S组和ACI组于造模后分为2、6、24、72 h时间点,每个时间点8只大鼠;取大鼠伤灶区脑组织测定Na＋-K＋-ATP酶和Ca2＋-ATP酶活性、TNF-α含量及脑组织含水量.结果 急性颅脑损伤后脑组织Na＋-K＋-ATP酶、Ca2＋-ATP酶活性降低（P〈0.05）,TNF-α含量升高（P〈0.05）;颅脑损伤后24 h脑水肿较严重.相关性分析提示,Ca2＋-ATP酶活性与TNF-α含量呈负相关（P〈0.05）.结论 急性颅脑损伤后可引起脑组织ATP酶活性降低、TNF-α含量增加,两者可能协同参与了急性颅脑损伤后脑水肿.     

Levels of Amyloid Beta-42, Interleukin-6 and Tumor Necrosis Factor-Alpha in Alzheimer’s Disease and Vascular Dementia

Amyloid β42 (Aβ42) and proinflammatory cytokines such as interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) have been suggested to contribute to the pathogenesis of Alzheimer's disease (AD) and vascular dementia (VaD). Our aim was to examine whether the changes in these parameters would be able to discriminate the patients with AD from those with VaD and from healthy individuals. We have analyzed the levels of Aβ42, IL-6 and TNF-α in the serum of newly diagnosed 28 AD patients, 16 VaD patients and 26 healthy non-demented controls. We also investigated whether there is an association between Aβ42, IL-6 and TNF-α levels and mini-mental state examination (MMSE) scores and body mass indexes (BMI) of patients. Our data showed a significant decrease in serum Aβ-42 levels in AD patients compared to VaD patients and controls. Levels of IL-6 and TNF-α were not different between AD patients, VaD patients and controls. We observed a correlation between Aβ-42 levels and MMSE scores and BMI levels in both AD and VaD patients. However, Aβ-42 levels were not correlated with IL-6 and TNF-α levels. Significantly lower levels of Aβ42 found in the serum of AD patients than that of VaD patients and controls suggests that it can be a specific biochemical marker for AD.     

Effect of Danshen aqueous extract on serum hs-CRP, IL-8, IL-10, TNF-α levels, and IL-10 mRNA, TNF-α mRNA expression levels, cerebral TGF-β1 positive expression level and its neuroprotective mechanisms in CIR rats

To observe the effects of Danshen aqueous extract (DSAE) on the cerebral tissue and nerve stem cells in cerebral ischemia reperfusion (CIR) rats. The model rats were prepared by occlusion of the middle cerebral artery for 2 h and then by reperfusion. They were randomly divided into five groups: a control group, an CIR group and three DSAE-treated groups. As compared with the sham control group, there was significant increase (P < 0.05, P < 0.01) in the serum high-sensitivity C-reactive protein (hs-CRP) and interleukin-8 (IL-8) levels, interleukin-10 (IL-10), tumor necrosis factor-alpha (TNF-α) levels, and IL-10 mRNA, TNF-α mRNA expression levels, function score, Infarct size, TUNEL + cell counts, cerebral transforming growth factor beta 1 (TGF-β1) positive expression and cerebral neuron specific enolase (NSE) levels, and decrease in fas-associated protein with death domain (FADD) and death-associated protein (Daxx) positive expression levels in the CIR group. Compared with CIR group, DSAE treatment dose-dependently significantly decreased serum hs-CRP, IL-8, IL-10, TNF-α levels, and IL-10 mRNA, TNF-α mRNA expression levels, function score, Infarct size, TUNEL + cell counts, cerebral TGF-β1 positive expression and cerebral NSE levels, and increase FADD and Daxx positive expression levels in the CIR + DSAE groups. Taken together, these results suggest that DSAE has a neuroprotective role in the CIR rats, which may be related to improvement of immunity function, proteins and genes expression.     

Levels of Porphyromonas gingivalis Fimbriae and Inflammatory Cytokines in Gingival Crevicular Fluid from Adult Human Subjects

The Porphyromonas gingivalis fimbriae level was examined in the gingival crevicular fluid (GCF) from adult human subjects using an immunoblot assay with a monoclonal antibody. The cytokines, interleukin-1α (IL-1α), IL-1β, IL-6 and tumor necrosis factor-a (TNF-α) levels in the GCF were quantified by enzyme-linked immunosorbent assay (ELISA). The reactivity of the GCF samples with the monoclonal antibody against P. gingivalis fimbriae was related to the IL-1β, IL-6 and TNF-α levels. Moreover, the fimbriae content was associated with the gingival index (GI). In contrast, no significant correlation was seen between the fimbriae content and IL-1α level. These results suggest that there are possible associations between P. gingivalis fimbriae and IL-1β, IL-6 and TNF-α in the gingival crevicular fluid.     

Picroside II Attenuates CCI-Induced Neuropathic Pain in Rats by Inhibiting Spinal Reactive Astrocyte-Mediated Neuroinflammation Through the NF-κB Pathway

Reactive astrocyte-mediated neuroinflammatory responses in the spinal dorsal horn have been reported to play a pivotal role in pathological pain. Chronic constriction injury (CCI) enhances the activation of nuclear factor kappa B (NF-κB), which is involved in neuropathic pain (NP). Picroside II (PII), a major active component of Picrorhiza scrophulariiflora, has been investigated for its anti-oxidative, anti-inflammatory, and anti-apoptotic activities. Here, we explored the analgesic effects of PII on a model of CCI-induced NP and investigated the levels of the GFAP protein and the mRNA and protein levels of pro-inflammatory cytokines in the spinal cord, including interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α). CCI significantly induced mechanical allodynia and thermal hyperalgesia. Intraperitoneal administration of PII remarkably reversed the CCI-induced mechanical allodynia and thermal hyperalgesia and reduced the mRNA and protein levels of IL-1β, IL-6, and TNF-α in the spinal cord. Additionally, according to the in vitro data, the PII treatment inhibited LPS-induced increases in the mRNA and protein levels of IL-1β, IL-6, and TNF-α and suppressed the NF-κB pathway by inhibiting the phosphorylation of NF-κB/p65 and the degradation of inhibitor of NF-κB (IκB) in astrocytes without toxicity to astrocytes. Overall, the analgesic effect of PII correlated with the inhibition of spinal reactive astrocyte-mediated neuroinflammation through the NF-κB pathway in rats with NP.     

Protective effects of salusin-α and salusin-β on renal ischemia/reperfusion damage and their levels in ischemic acute renal failure

Salusin-α and salusin-β are expressed in many tissues including the central nervous system, vessels and kidneys; they have been shown to decrease endoplasmic reticulum stress during heart ischemia/reperfusion (I/R) and to decrease apoptosis. We investigated the relation of salusin-α and salusin-β levels to acute ischemic renal failure. We also investigated whether these peptides are protective against renal I/R damage. Fifty-three rats were divided into six groups: control, I/R, I/R + salusin-α1, I/R + salusin-α10, I/R + salusin-β1 and I/R + salusin-β10. After removing the right kidney, the left kidney was subjected to ischemia for 1 h and reperfusion for 23 h. The treatment groups were injected subcutaneously at the beginning of ischemia with 1 or 10 μg/kg salusin-α, and 1 or 10 μg/kg salusin-β. Histopathology was assessed at the end of the experiment. Superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-PX) activity and malondialdehyde (MDA) levels were measured in the kidney tissue. Serum levels of blood urea nitrogen (BUN), creatinine (Cre), tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and interleukin-1 beta (IL-1β) also were measured. Levels of salusin-α and salusin-β were measured in the serum and kidney tissues of the control and I/R groups. SOD, CAT and GSH-PX activities were decreased and the levels of MDA, TNF-α, IL-6, IL-1β, BUN and Cre were increased in the I/R group compared to controls. Severe glomerular and tubular damage was apparent in the I/R group compared to controls. The level of salusin-β was decreased in the serum and kidney tissue of the I/R group compared to controls, whereas the level of salusin-α was decreased in the serum and increased in the kidney tissue. Salusin-α and salusin-β administration increased SOD and GSH-PX enzyme activation and decreased the levels of MDA, TNF-α, IL-6 and IL-1β compared to the I/R group. BUN and Cre levels were decreased in the I/R + salusin-α1 group and the level of Cre was decreased in I/R + salusin-β10 group compared to the I/R group. We demonstrated a protective effect of salusin-α and salusin-β against renal I/R damage. Changes in the levels of salusin-α and salusin-β in the I/R group suggest that these peptides may be associated with acute renal failure.     

Propofol Administration Modulates AQP-4 Expression and Brain Edema After Traumatic Brain Injury

The increased intracranial pressure caused by brain edema following traumatic brain injury (TBI) always leads to poor patient prognosis. Aquaporin-4 (AQP-4) plays an important role in edema formation and resolution, which may provide a novel therapeutic target for edema treatment. In this present study, we found that propofol treatment, within a short time, after TBI significantly reduced brain edema in a controlled cortical injury rat model and suppressed in vivo expression of AQP-4. The ameliorating effect of propofol was associated with attenuated expression of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α). In addition, the regulatory effect of propofol on AQP-4 expression was investigated in cultured astrocytes. Results showed that propofol could block the stimulatory effect of IL-1β and TNF-α on AQP-4 expression in cultured astrocytes. We also found that both NFκB and p38/MAPK pathways were involved in IL-1β and TNF-α-induced AQP-4 expression and that propofol functions as a dual inhibitor of NFκB and p38/MAPK pathways. In conclusion, treatment with propofol, within a short time, after TBI attenuates cerebral edema and reduces the expression of AQP-4. Propofol modulates acute AQP-4 expression by attenuating IL-1β and TNF-α expression and inhibiting IL-1β and TNF-α induced AQP-4 expression.     

膝关节炎患者关节置换术后细菌感染严重程度与IL-1β、TNF-α、IL-6水平的相关性

目的探讨膝关节炎患者关节置换术后细菌感染严重程度与IL-1β、TNF-α、IL-6水平的相关性。方法选取2016年8月至2018年8月于我院进行治疗的70例膝关节炎关节置换术后细菌感染患者作为试验组,参照Michel Lequesen推荐的膝关节炎严重性判断标准将膝关节炎关节置换术后细菌感染患者分为极严重组、非常严重组、严重组、中度组和轻度组。选取同期来我院体检中心进行体检的健康者50例为对照组。采用全自动生化免疫分析仪测定血清IL-1β、IL-6及TNF-α水平。结果试验组患者血清IL-1β、TNF-α、IL-6水平均明显高于对照组(均P<0.05)。极严重组患者血清IL-1β、TNF-α、IL-6水平显著高于非常严重组、严重组、中度组和轻度组(均P<0.05)。非常严重组患者血清IL-1β、TNF-α、IL-6水平显著高于严重组、中度组和轻度组(均P<0.05)。严重组患者血清IL-1β、TNF-α、IL-6水平显著高于中度组和轻度组(均P<0.05)。IL-1β、TNF-α、IL-6水平与膝关节炎关节置换术后细菌感染严重程度呈显著正相关(均P<0.05)。结论膝关节炎关节置换术后细菌感染患者血清IL-1β、TNF-α、IL-6水平明显升高,并且病情越严重,IL-1β、TNF-α、IL-6水平越高。     

Relationship between anti-fibrotic effect of Panax notoginseng saponins and serum cytokines in rat hepatic fibrosis

The aim of this study was to investigate the relationship between anti-fibrotic effect of Panax notoginseng saponins (PNS) and serum cytokines in rat hepatic fibrosis. Hepatic fibrosis induced by carbon tetrachloride (CCl₄) was studied in animal models using SD rats. Liver index, serum alanine amino transferase (ALT), aspartate amino transferase (AST), transforming growth factor-β1 (TGF-β1), tumour necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and interleukin-10 (IL-10) were measured, respectively. Liver index and the degree of liver fibrosis were also determined. Our results showed that the levels of ALT, AST and liver index in PNS-treated group were markedly lower than those in model group. PNS therapy also significantly attenuated the degree of hepatic fibrosis, collagen area and collagen area percent in liver tissue. Furthermore, the levels of serum TGF-β1, TNF-α and IL-6 were strikingly reduced in PNS-treated group compared with model group while the production of IL-10 was up-regulated. These findings demonstrate that PNS has certain therapeutic effects on hepatic fibrosis probably by immunoregulating the imbalance between pro-fibrotic and anti-fibrotic cytokines.     

Ghrelin attenuates brain injury after traumatic brain injury and uncontrolled hemorrhagic shock in rats

Traumatic brain injury (TBI) and hemorrhagic shock often occur concomitantly due to multiple injuries. Gastrointestinal dysfunction occurs frequently in patients with TBI. However, whether alterations in the gastrointestinal system are involved in modulating neuronal damage and recovery after TBI is largely neglected. Ghrelin is a "gut-brain" hormone with multiple functions including antiinflammation and antiapoptosis. The purpose of this study was to determine whether ghrelin attenuates brain injury in a rat model of TBI and uncontrolled hemorrhage (UH). To study this, brain injury was induced by dropping a 450-g weight from 1.5 m onto a steel helmet attached to the skull of male adult rats. Immediately after TBI, a midline laparotomy was performed and both lumbar veins were isolated and severed at the junction with the vena cava. At 45 min after TBI/UH, ghrelin (4, 8 or 16 nmol/rat) or 1 mL normal saline (vehicle) was intravenously administered. Brain levels of TNF-α and IL-6, and cleaved PARP-1 levels in the cortex were measured at 4 h after TBI/UH. Beam balance test, forelimb placing test and hindlimb placing test were used to assess sensorimotor and reflex function. In additional groups of animals, ghrelin (16 nmol/rat) or vehicle was subcutaneously (s.c.) administered daily for 10 d after TBI/UH. The animals were monitored for 28 d to record body weight changes, neurological severity scale and survival. Our results showed that ghrelin downregulated brain levels of TNF-α and IL-6, reduced cortical levels of cleaved PARP-1, improved sensorimotor and reflex functions, and decreased mortality after TBI/UH. Thus, ghrelin has a great potential to be further developed as an effective resuscitation approach for the trauma victims with brain injury and severe blood loss.     

Eupatilin alleviates inflammatory response after subarachnoid hemorrhage by inhibition of TLR4/MyD88/NF-κB axis

Early brain injury (EBI) is associated with the adverse prognosis of subarachnoid hemorrhage (SAH) patients. The key bioactive component of the Chinese herbal medicine Artemisia asiatica Nakai (Asteraceae) is eupatilin. Recent research reports that eupatilin suppresses inflammatory responses induced by intracranial hemorrhage. This work is performed to validate whether eupatilin can attenuate EBI and deciphers its mechanism. A SAH rat model was established by intravascular perforation in vivo. At 6 h after SAH in rats, 10 mg/kg eupatilin was injected into the rats via the caudal vein. A Sham group was set as the control. In vitro, BV2 microglia was treated with 10 μM Oxyhemoglobin (OxyHb) for 24 h, followed by 50 μM eupatilin treatment for 24 h. The SAH grade, brain water content, neurological score, and blood-brain barrier (BBB) permeability of the rats were measured 24 h later. The content of proinflammatory factors was detected via enzyme-linked immunosorbent assay. Western blot analysis was conducted to analyze the expression levels of TLR4/MyD88/NF-κB pathway-associated proteins. In vivo, eupatilin administration alleviated neurological injury, and decreased brain edema and BBB injury after SAH in rats. Eupatilin markedly reduced the levels of interleukin-1β (IL-1β), IL-6, and tumor necrosis factor-α (TNF-α), and suppressed the expression levels of MyD88, TLR4, and p-NF-κB p65 in the SAH rats' cerebral tissues. Eupatilin treatment also reduced the levels of IL-1β, IL-6, and TNF-α, and repressed the expression levels of MyD88, TLR4, and p-NF-κB p65 in OxyHb-induced BV2 microglia. Additionally, pyrrolidine dithiocarbamate or resatorvid enhanced the suppressive effects of eupatilin on OxyHb-induced inflammatory responses in BV2 microglia. Eupatilin ameliorates SAH-induced EBI via modulating the TLR4/MyD88/NF-κB pathway in rat model.     

小檗碱对大鼠脑缺血/再灌注损伤的保护作用及免疫机制*

目的:探讨小檗碱对大鼠脑缺血/再灌注损伤的保护作用及免疫机制。方法:50只SD大鼠随机分为假手术组(Sham group)、模型组(Model group)、小檗碱低剂量组(BBR-L,25 mg/kg)、小檗碱中剂量组(BBR-M,50 mg/kg)、小檗碱高剂量组(BBR-H,100 mg/kg),每组各10只。采用Longa线栓法建立脑缺血/再灌注大鼠模型,缺血2 h后再灌注24 h处理。于造模成功2 h后灌胃给药,假手术组和模型组组按上述方法同体积给予生理盐水。给药24 h后,测定各组大鼠神经功能缺损程度评分及脑梗死率;采用ELISA法检测抗氧化酶SOD和GSH-Px的活性、细胞因子TNF-α、IFN-β、IL-6和NO的含量;采用流式细胞术检测CD4⁺、CD8⁺及CD4⁺/CD8⁺血清含量;进一步采用RT-qPCR与Western blot技术检测大鼠脑组织内NF-κB-NLRP3信号轴关键基因及蛋白的表达情况。结果:与假手术组比较,模型组大鼠神经功能缺损程度、脑梗死率均升高(P＜0.05),且血清NO、TNF-α、IFN-β、IL-6含量和脑组织的NF-κB p65、NLRP3、ASC及caspase-1基因与蛋白表达水平均升高(P＜0.05),而血清中SOD、GSH-Px活性和CD4⁺、CD8⁺及CD4⁺/CD8⁺水平下降(P＜0.05);与模型组比较,BBR-H、BBR-M、BBR-L组大鼠神经功能缺损程度、脑梗死率均下降(P＜0.05),且血清NO、TNF-α、IFN-β、IL-6含量和脑组织的NF-κB p65、NLRP3、ASC及caspase-1基因与蛋白表达水平均降低(P＜0.05),而血清中SOD、GSH-Px活性和CD4⁺、CD8⁺及CD4⁺/CD8⁺水平升高(P＜0.05)。结论:小檗碱可通过减轻氧化应激,抑制炎症反应,增强免疫功能,减轻大鼠脑缺血/再灌注损伤,其机制可能与抑制NF-κB-NLRP3信号有关。