Dexamethasone treatment modulates aquaporin-4 expression after intracerebral hemorrhage in rats

This study investigated whether dexamethasone (DEX) treatment could regulate the expression of aquaporin-4 (AQP4) in rats with intracerebral hemorrhage (ICH). The results demonstrated that DEX significantly reduced AQP4 mRNA level in the perihematomal area compared with control group, but it increased the level in the brain area surrounding the third ventricle at day 1 post-ICH. There was no difference in AQP4 protein levels between DEX group and control group at the two above-mentioned brain regions at day 1 after ICH. The changes in AQP4 protein induced by DEX were marked at day 3 following surgery and still lasted at day 5 post-ICH, which were accompanied by a reduction of brain edema. Our results demonstrated that the expression of AQP4 protein after ICH was region-specific, time-dependent, and also indicated that DEX-induced cerebral edema clearance was correlated with the regulation of AQP4 expression in different brain regions.     

长春西汀减轻脑出血大鼠脑组织的炎症损伤

目的:探讨长春西汀对脑出血大鼠的干预作用以及对炎症损伤的影响。方法:将大鼠随机分为假手术组、脑出血组以及长春西汀低剂量组、中剂量组和高剂量组。除假手术组外,其余大鼠均通过注射VII型胶原酶的方法建立脑出血模型,长春西汀低剂量组、中剂量组和高剂量组分别给予腹腔注射0.5、1.0和1.5 mg/kg长春西汀,每天1次,共给药7 d。给药完成后,对各组大鼠进行神经功能缺损评分,称重法计算脑组织的含水量,检测脑组织中髓过氧化物酶(MPO)活性,Western blot法检测脑组织中Toll样受体4(TLR4)、核因子κB(NF-κB)、细胞间黏附分子1(ICAM-1)与血管细胞黏附分子1(VCAM-1)的蛋白表达。结果:与脑出血组相比,给予长春西汀干预的大鼠神经功能缺损评分显著下降(P0.05),脑组织含水量明显下降(P0.05),MPO活性显著降低(P0.05),脑组织中TLR4、NF-κB、ICAM-1与VCAM-1的蛋白表达显著下降(P0.05)。结论:长春西汀对脑出血大鼠脑组织具有保护作用,可能是通过抑制TLR4诱导的NF-κB信号通路以及ICAM-1与VCAM-1的表达来抑制炎症反应。     

AQP4,MMP-2及MMP-9在高血压性脑出血后脑水肿中的早期表达

目的：探讨水通道蛋白4（AQP4）及基质金属蛋白酶2，9（MMP-2，MMP-9）表达在高血压性脑出血后早期脑水肿发生中的作用。方法：应用免疫组织化学方法检测高血压性脑出血患者出血后24h内血肿周围脑组织中AQP4及MMP-2、MMP-9的表达。结果：高血压性脑出血患者血肿周围有明显的脑动脉硬化和脑水肿发生，正常对照组中未发现明显的MMP-2，MMP-9阳性表达，AOP4可见少量表达，而在高血压性脑出血组出血24h内AQP4，MMP-2及MMP-9则见明显表达（P〈0．01），尤其是AQP4的阳性表达升高更为显著，在小血管及神经胶质细胞中均见大量表达。结论：AQP4、MMP-2及MMP-9的早期上调表达与高血压性脑出血后脑水肿的发生具有密切的关系。尤其是AQP4的表达在早期脑水肿的发生、发展中具有重要的作用。     

黄连素对实验性脑出血大鼠脑水肿与神经功能的影响

目的:研究不同剂量黄连素对实验性脑出血(intracerebral hemorrhage,ICH)大鼠脑水肿与神经功能的影响,并初步探讨其作用机制.方法:实验共分为假手术组(Sham组),脑出血组(ICH组),药物低(Ber-10组)、中(Ber-20组)、高剂量组(Ber-40组),每组10只,除假手术组采用注射胶原酶的方法建立实验性脑出血大鼠模型,药物低、中、高剂量组分别给予黄连素10,20或40 mg/kg灌胃,每天1次,共给药28 d;对各组大鼠进行神经功能缺损评分,干湿重法测定脑含水量,Tunel法检测血肿周围脑组织细胞凋亡数,Western印迹法检测Cleaved cas-3,Cleaved cas-9,线粒体与胞浆中cyto-c的蛋白表达水平.结果:与脑出血组大鼠相比,采用黄连素给药后大鼠的神经功能缺损评分明显上升(P<0.05),脑含水量明显下降(P<0.05);血肿周围脑组织细胞凋亡数显著降低(P<0.05),Cleaved cas-3和Cleaved cas-9的蛋白表达,线粒体中cyto-c的蛋白表达明显上升(P<0.05)而胞浆中cyto-c的蛋白表达明显下降(P<0.05).结论:黄连素具有改善实验性脑出血大鼠神经功能缺损以及保护脑细胞的作用,可能与黄连素抑制线粒体凋亡通路有关.     

脑出血大鼠大脑颞顶叶皮质水通道蛋白-4的表达

目的：研究脑出血脑水肿时，健侧大脑半球颞顶叶皮质水通道蛋白-4(AQP-4)表达变化及机制。 方法： 采用立体定向注射无肝素自体血制作大鼠脑出血模型,用MRI T2加权和病理学检查测定双侧半球颞顶叶皮质的水肿程度，RT-PCR和Western blotting检测两个部位AQP-4 mRNA和蛋白质的表达变化，最后从超微结构的角度探讨健侧半球颞顶叶皮质AQP-4改变的原因。 结果： 脑出血模型组大鼠出血侧和健侧颞顶叶皮质AQP-4表达均高于假手术组（P<0.05），脑出血1 d AQP-4 mRNA和蛋白即明显加强,第3 d达峰值,其后逐渐有所下降,但持续1周仍高于假手术组。健侧AQP-4表达弱于出血侧，但仍高于假手术组。电镜观察结果：出血侧颞顶叶皮质出现神经元水样变性，健侧颞顶叶皮质神经细胞核膜间隙无扩张，核周体无明显空泡形成，但线粒体明显肿胀呈球形,部分嵴断裂或溶解消失,透明成空泡;粗面内质网扩张,表面核糖体出现脱落，提示该部位细胞水肿不明显，但是出现了缺氧性改变。 结论： 脑出血后健侧颞顶叶皮质AQP-4在水肿不明显的情况下出现表达增强，其增高可能与脑缺氧有关。     

Combined neuroprotective effects of celecoxib and memantine in experimental intracerebral hemorrhage

Memantine, a N-methyl-D-aspartate (NMDA) receptor antagonist, inhibits hematoma expansion and celecoxib, a selective cyclooxygenase-2 (COX-2) inhibitor, reduces perihematomal inflammation in intracerebral hemorrhage. We examined whether the combination treatment has additive effects in experimental intracerebral hemorrhage (ICH). ICH was induced using stereotaxic infusion of collagenase into brains of adult rats. After the induction of ICH, rats were treated with intraperitoneal injection of memantine (20 mg/kg), celecoxib (20 mg/kg) or both agents. Only vehicles were administrated in rats of the control group. Results showed that the combination treatment of memantine and celecoxib reduced both hematoma volume and brain edema. Combination treatment also induced the better functional recovery with further attenuation of cerebral inflammation and apoptosis compared to the control group. When compared to the single agent groups, the combination treatment showed better effects in neuroprotection and anti-inflammation. These results suggest the feasible combined application of memantine and celecoxib in ICH treatment.     

Internalization of Aquaporin‐4 After Collagenase‐Induced Intracerebral Hemorrhage

Brain edema formation following intracerebral hemorrhage (ICH) appears to be related with aquaporin‐4 (AQP4), which is critically involved in brain volume homeostasis and water balance. Despite its importance, the regulation of AQP4 expression involved in transmembrane water movements still remains rudimentary. Many studies suggest that the internalization of several membrane‐bound proteins, including AQP4, may occur with or without lysosomal degradation. Previously, we investigated the internalization of AQP4 in retinal ischemic‐reperfusion model. Here, we test the hypothesis that AQP4 is internalized post‐ICH and then degraded in the lysosome. The results demonstrated that both AQP4 and the mannose‐6‐phosphate receptor (MPR) co‐localized in perihematomal region at 6 hr post‐ICH. In addition, AQP4 and lysosomal‐associated membrane protein 1 (LAMP1) also co‐localized in perihematomal region, with co‐expression increasing followed by a gradual decrease at different time windows post‐ICH (6, 12, 24, 48, and 72 hr). After ICH, the Evans blue leakage happened very early at 1 hr and the brain swelling occurred at 3 hr. Moreover, we also found the AQP4 mRNA and AQP4 protein were increased post‐ICH. These results suggest that AQP4 is internalized and the lysosome is involved in degrading the internalized AQP4 post‐ICH. Both the AQP4 internalization and lysosomal degradation may provide biophysical insights regarding the potential of new treatments for brain edema. Anat Rec, 298:554–561, 2015. © 2014 Wiley Periodicals, Inc.     

大鼠脑出血后V1αR和AQP4表达升高与脑水肿的形成相关

 目的 研究精氨酸加压素1α受体（V1αR）和水通道蛋白4（AQP4）在SD大鼠脑出血（ICH）脑组织中的表达变化及两者在脑水肿形成中的作用。方法 将自体血注入大鼠尾状核建立脑出血模型,分别于术后6、12、24和48h运用干湿重法、免疫荧光和Western blot检测脑含水量(BWC)、V1αR和AQP4蛋白的表达。结果 脑出血后6h,V1αR和AQP4的表达开始增加,至48h达到高峰分别为（21.88±0.44和23.16±0.67）,显著高于假手术组（14.32±0.55和13.90±0.40）。与脑含水量呈正相关（P＜0.05）。结论 V1aR介导的AQP4表达升高是脑出血后脑水肿形成的主要原因。     

脑出血后水通道蛋白4与内向整流性钾通道4.1的再分布与脑水肿形成的关系

目的:探讨水通道蛋白4(AQP4)与内向整流性钾通道4.1(Kir4.1)在出血性脑水肿形成中的作用.方法:采用胶原酶注入法建立脑出血(ICH)模型,运用干湿重法、伊文氏蓝(EB)测定法、荧光双标和RT-PCR分别检测ICH后脑水含量(BWC)、血脑屏障(BBB)通透性、AQP4与Kir4.1及两者mRNA的表达变化,并对每一时间点AQP4 mRNA、 Kir4.1 mRNA表达量做比值分析.结果:ICH后3h BWC已明显增高,至48h达到峰值;EB含量在ICH后48h内高于对照组,以6h最甚;荧光双标显示,AQP4与Kir4.1强弱不等地表达在双侧软脑膜、室管膜、脉络膜等与水代谢密切相关的界面上.ICH后6h及其以后时段,两者的mRNA均明显增高,且于48h达到峰值;比值分析显示,ICH后6h及以后时段,AQP4 mRNA的增加量明显大于Kir4.1 mRNA的增加量.结论:ICH早期的脑水肿主要由血脑屏障通透性增加所致,而ICH中后期的脑水肿主要由AQP4与Kir4.1的再分布引起.     

Ghrelin对脑缺血再灌注大鼠脑水肿和水通道蛋白4表达的影响

 目的： 探讨ghrelin对脑缺血再灌注大鼠脑水肿、血脑屏障通透性及水通道蛋白4（AQP4）表达的影响。方法： 成年SD雄性大鼠随机分为3组:sham组、大脑中动脉阻塞（MCAO）组和ghrelin处理组。采用线栓法复制MCAO模型（缺血2 h,再灌注22 h）。Ghrelin组大鼠于再灌开始时经股静脉注射ghrelin 10 nmol/kg。TTC染色观察脑梗死体积,神经功能评分判断脑功能障碍程度,分别以体积计算和干湿重法检测脑肿胀程度和脑含水量的变化,收集脑血管外伊文思蓝(EB)来评估血脑屏障的破坏程度,免疫组化和Western blotting检测AQP4的表达变化。结果： 与MCAO组比较,ghrelin处理组的脑梗死体积较小（P<0.01）,神经功能评分较低（P<0.01）,脑组织中的EB渗出量较少（P<0.01）。Ghrelin处理组大鼠的脑肿胀体积、脑含水量和AQP4表达明显低于MCAO组（P<0.05）。结论： Ghrelin减轻大鼠脑缺血/再灌注损伤,减轻脑水肿和血脑屏障的破坏,抑制脑组织中AQP4的表达。AQP4在ghrelin的脑保护机制中可能发挥重要作用。     

Baicalin Attenuates Brain Edema in a Rat Model of Intracerebral Hemorrhage

Baicalin is a flavonoid compound purified from the roots of Scutellaria baicalensis, which possesses multiple biological activities. Previous studies have shown that baicalin is protective in ischemic cerebral diseases. The aim of the present study was to examine the effects of baicalin on brain injury in a rat model of intracerebral hemorrhage (ICH) and to explore the possible mechanisms. Intracerebral hemorrhage was induced in male Wistar rats by injection of 0.5 U collagenaseVII to the caudate nucleus. Sham operation rats were injected with equal volume of saline. After the induction of ICH, the rats were randomly divided into four groups and administered with different dose of baicalin (0, 25, 50, or 100 mg/kg in saline) through peritoneal injection. The brain tissues around the hemorrhage areas were collected on days 1, 3, and 5 after treatment. Brain edema was analyzed by desiccation method; the metalloproteinase-9 (MMP-9) protein and mRNA expression were determined by western blotting and real time RT-PCR, respectively. Nuclear factor-κB (NF-κB) protein expression was analyzed by western blotting. IL-1β and IL-6 levels were determined by enzyme-linked immunosorbent assay. Blood–brain barrier permeability was determined by Evans blue leakage method. The results showed that baicalin reduced brain edema following ICH in a dose-dependent manner, with concomitant inhibition of NF-κB activation and suppression of MMP-9 expression. In addition, baicalin also reduced IL-1β and IL-6 production, as well as blood–brain barrier permeability. The above results indicated that baicalin prevents against perihematomal edema development after intracerebral hemorrhage possibly through an anti-inflammatory mechanism.     

依达拉奉对急性脑缺血/再灌注大鼠AQP4和Aβ表达及MMP-2和MMP-9活性的影响

目的:探讨依达拉奉对急性脑缺血/再灌注大鼠脑损伤的治疗效果及其作用机制。方法:选取健康雄性SPF级SD大鼠,采用随机数字表法分为假手术组(等体积生理盐水)、模型组(造模后给予等体积生理盐水)、低剂量组(依达拉奉6 mg/kg)和高剂量组(依达拉奉10 mg/kg),采用Zea Longa线栓法造模,并对各组大鼠在术后的神经功能评分,TTC法测定脑梗死体积;采用RT-qPCR法检测脑组织中水通道蛋白4(aquaporin 4,AQP4)和β-淀粉样蛋白(amyloidβ-protein,Aβ)的mRNA表达; Western blot法检测Aβ及AQP4的蛋白水平;采用明胶酶谱法检测基质金属蛋白酶2(matrix metalloproteinase 2,MMP2)和MMP9的活性。结果:相对于模型组,依达拉奉能显著减轻大鼠神经功能障碍、组织学损伤和脑水肿,降低AQP4和AβmRNA和蛋白水平及MMP2和MMP9活性,且高剂量组效果优于低剂量组(P 0. 05)。结论:依达拉奉减轻急性缺血性脑卒中大鼠脑组织损伤的机制可能与其抑制AQP4和Aβ表达及MMP2和MMP9活性有关。     

NF-κB抑制剂对脑出血大鼠神经损伤的影响

目的:探讨核因子κB(NF-κB)抑制剂吡咯烷二硫代氨基甲酸盐(PDTC)对脑出血(ICH)大鼠神经功能及神经细胞凋亡的调节作用。方法:取SPF级SD大鼠随机分假手术组(sham组)、ICH组、PDTC低浓度组(Plow组)和PDTC高浓度组(P_(high)组),每组6只。采用自体血注入法建立大鼠ICH模型,Plow组和P_(high)组在缺血后2h分别给予100 mg/kg和200 mg/kg的PDTC腹腔注射,sham组和ICH组给予等体积的生理盐水;24 h后,采用改良的Longa分级法进行神经功能评分;TUNEL法测定神经细胞凋亡情况;并检测脑组织中丙二醛(MDA)的含量和超氧化物歧化酶(SOD)的活性。此外,采用Western blot法检测脑组织中p-P65及cleaved caspase-3的蛋白水平。结果:与sham组相比,ICH组大鼠的神经功能评分显著升高(P0.05);应用PDTC后,Plow组和P_(high)组动物的神经功能评分都有所降低,但2组间差异无统计学意义。与sham组相比,ICH组大鼠神经细胞凋亡明显增多(P0.05);应用PDTC后,2组动物神经细胞凋亡数目显著降低,且P_(high)组的细胞凋亡数目显著低于Plow组(P0.05)。与sham组相比,ICH组大鼠脑组织的MDA含量升高,SOD活性降低(P0.05);应用PDTC后,2组动物脑组织中的MDA含量显著降低,而SOD活性升高,且这一趋势在P_(high)组表现更为明显。与sham组相比,ICH组大鼠脑组织中p-P65和cleaved caspase-3的蛋白水平显著增加(P0.05);应用PDTC后,2组动物脑组织中p-P65和cleaved caspase-3的蛋白水平显著降低,且P_(high)组中p-P65和cleaved caspase-3的蛋白水平显著低于Plow组。结论:NF-κB抑制剂PDTC能减轻脑出血后继发性脑损伤,且大剂量作用效果较好;其作用机制可能与降低MDA含量、提高SOD活性进而抑制神经细胞凋亡有关。     

呋塞米对脑出血模型大鼠PERK/eIF2α/CHOP通路及继发性脑损伤的影响

目的探讨呋塞米(FUR)对脑出血(ICH)大鼠RNA依赖的蛋白激酶样内质网激酶(PERK)/真核起始因子2α(eIF2α)/转录因子CCAAT增强子结合蛋白的同源蛋白(CHOP)通路及继发性脑损伤的影响。方法将大鼠分为假手术组、模型组(脑定位注射Ⅳ型胶原酶,建立ICH模型)、FUR低剂量组(FUR-L)、FUR中剂量组(FUR-M)、FUR高剂量组(FUR-H)、神经节苷脂组,每组18只。药物处理后,检测神经功能,对其进行Longa评分;测量大脑含水量;用Evans蓝外渗实验检测血脑屏障损伤;用HE染色检测海马神经元损伤;用酶联免疫吸附法(ELISA)检测血清γ干扰素(IFN-γ)、白细胞介素-6(IL-6)水平;用免疫印迹法检测大鼠脑组织PERK/eIF2α/CHOP通路相关蛋白p-PERK/PERK、p-eIF2α/eIF2α、CHOP表达。结果相比假手术组,模型组大鼠海马神经元变性坏死,皱缩变小,数目明显减少,呈现严重病理损伤,Longa评分、Evans蓝渗出量、大脑含水量、血清IFN-γ及IL-6水平、p-PERK/PERK、p-eIF2α/eIF2α、CHOP表达水平明显升高...     

Proteasomal inhibition in intracerebral hemorrhage: neuroprotective and anti-inflammatory effects of bortezomib

Inflammation is an important pathophysiologic mechanism of injury induced by intracerebral hemorrhage (ICH). The ubiquitin-proteasome system (UPS) regulates the inflammatory responses via the up-regulation of several pro-inflammatory molecules. In this study, we determined that a potent proteasome inhibitor, bortezomib, exerted therapeutic effects in experimental model of ICH. Either bortezomib (0.05, 0.2, 0.5, 1mg/kg) or vehicle was intravenously administered 2h after ICH induction. The high doses of bortezomib caused high mortality rates. Bortezomib at 0.2 mg/kg reduced the early hematoma growth and alleviated hematoma volume and brain edema at 3 days after ICH, compared with the ICH-vehicle group. The numbers of myeloperoxidase(+) neutrophils, Ox42(+) microglia, and TUNEL(+) cells in the perihematomal regions were decreased by bortezomib. Bortezomib induced significant decrements of mRNA expression of TNF-alpha and IL-6. The production of iNOS and COX2 was also reduced significantly by bortezomib. We concluded that the early treatment with bortezomib induced a reduction in the early hematoma growth and mitigated the development of brain edema, coupled with a marked inhibitory effect on inflammation in ICH.     

米诺环素对大鼠脑出血后VEGF、GLUT-1 表达的影响

目的 研究米诺环素对脑出血大鼠VEGF、GLUT-1表达的影响,探讨其的脑保护作用机制。方法 选用雄性Wistar大鼠60只,随机分为假手术组、模型组及米诺环素组,每组20只。采用Ⅶ型胶原酶法制备Wistar大鼠右侧纹状体出血模型。模型制备成功后米诺环素组给予腹腔注射米诺环素,剂量为22.5 mg/kg,2次/d,假手术组和模型组给予腹腔注射等量的生理盐水。观察脑缺血后改良神经功能缺损评分（mNSS）、脑组织含水量,Western blots及免疫组化法分别检测VEGF及GLUT-1的表达。结果 与模型组相比,米诺环素组大鼠脑组织的mNSS评分、脑组织含水量、VEGF及GLUT-1的表达量降低,差异均有统计学意义（P＜0.05）。结论 米诺环素可显著减轻脑出血后大鼠的神经功能损害及组织学损伤,并提高大鼠脑组织VEGF及GLUT-1的表达水平,对脑出血具有一定的神经保护作用,其机制可能与增强VEGF及GLUT-1的表达有关。     

黄芩茎叶总黄酮对大鼠大脑海马区微血管和血脑屏障缺血再灌注损伤的预防作用

目的 探讨黄芩茎叶总黄酮（SSTF）对脑缺血再灌注海马区微血管、血脑屏障（BBB）损伤的预防性保护作用。 方法 SD大鼠90只,随机分为假手术组（sham组）、模型组（IR组）和SSTF预处理组,造模前1周SSTF各组灌胃给药,低、中、高剂量组分别给50、100、200 mg/(kg·d),共7d。IR组和SSTF各组制作脑缺血再灌注模型,术后评价神经功能缺损变化,干湿重法和伊文思蓝法检测脑组织含水量和微血管通透性,单宁酸 氯化铁媒染（TA-Fe）法显示并观测大鼠微血管密度(MVD)和微血管面积比(MVA),Real-time PCR法检测水通道蛋白4（AQP4） mRNA的表达水平,电镜观察血脑屏障完整性。 结果 SSTF各组与IR组比,神经功能缺损评分、脑组织含水量、微血管通透性明显降低（P＜0.01,P＜0.05）,MVD、MVA增加（P＜0.01）, AQP4 mRNA表达增强(P＜0.01),BBB损伤不同程度减轻,内皮细胞肿胀渐消退,紧密连接松解好转,基膜渐连续、清晰,胶质细胞足板胞质溶解减轻,渐接近正常;SSTF 中、高组上述表现较SSTF 低组好转的更明显（P<0.01）,SSTF 中组与SSTF 高组比较差异无统计学意义（P＞0.05）。 结论 SSTF干预对海马区微血管、血脑屏障和神经损伤有预防性保护作用,其作用机制可能是通过增加有效微血管再通数量,维持血脑屏障完整和功能,减轻脑水肿实现的。SSTF的有效预防剂量为100mg/(kg·d)。     

两种脑出血动物模型的AQP4表达与磁共振成像的比较研究

目的比较两种脑出血(ICH)动物模型磁共振成像(MRI)的差异,并对其分子机制进行探讨。方法将胶原酶和自体血注入大鼠尾状核建立脑出血模型,分别于术后6h、12h、1d、2d、3d和7d进行磁共振成像扫描,观察两组脑出血模型血肿体积的大小;运用干湿重法、伊文思蓝(EB)测定法和免疫印迹分别检测两组脑出血模型的脑含水量(BWC)、血脑屏障(BBB)通透性和AQP4的表达变化。结果胶原酶模型组的血肿体积在脑出血后1d达到最大并持续至3d,自体血组的血肿体积在12h达到最大;胶原酶模型组的BWC在1d达到高峰并持续至3d,而自体血组BWC在2d逐渐降低;胶原酶模型组的EB含量在12h达到高峰并持续至2d,而自体血组EB含量在1d逐渐下降;两组模型的AQP4表达量在6h开始升高,1d达到高峰,2d逐渐降低。两组模型比较具有统计学差异(P<0.05)。结论胶原酶对BBB和细胞外基质的破坏以及AQP4的表达变化是两组脑出血模型血肿体积出现差异的重要原因。     

Effects of intracerebral hemorrhage and subsequent minimally invasive hematoma aspiration on expression of apoptosisrelated genes in rats

This study was to investigate the effects of intracerebral hemorrhage (ICH) and subsequent minimally invasive hematoma aspiration on the expression of apoptosis-related genes in rats. IV-collagenase was injected to the caudate nucleus of the rats to make ICH models. In the control group, 30 Sprague-Dawley (SD) rats were mock treated with saline instead of collagenase. Thirty SD rats with successful modeling were designated as the ICH group. Twenty-five SD rats with successful modeling and subsequent minimally invasive hematoma aspiration were designated as the therapy group. Expression of heat shock protein 70 (Hsp70), B-cell leukemia/lymphoma 2 (Bcl-2) and Bcl-2-associated X protein (Bax) in the brain tissues was detected by immunohistochemical assays. The expression of Hsp70, Bcl-2 and Bax in the control group was very low, and significantly increased in the ICH group and the therapy group. At each indicated time point, Hsp70 expression in the therapy group was significantly lower than that of the ICH group, Bax expression in the therapy group was significantly lower than that of the ICH group and Bcl-2 expression in the therapy group was significantly higher than that of the ICH group. These results suggest that ICH led to increased expression of apoptosis-related genes in the brain tissues. Hematoma aspiration up-regulated ICH induced Bcl-2 expression while down-regulated ICH induced Hsp70 and Bax expression.     

HET0016在小鼠纹状体脑出血损伤中的作用及其可能机制

目的研究20-羟二十烷四烯酸(20-HETE)抑制剂HET0016对脑缺血的作用及其可能机制。方法选择C57品系小鼠,制备胶原酶诱导的纹状体脑出血模型。阻断剂组予以20-HETE的阻断剂HET00162mg/kg,于脑出血模型制备后2h腹腔注射。对照组予以等体积溶剂羟丙基-BETA-环糊精腹腔注射。各组分别于脑出血后各时点取材,测定脑损伤体积,脑含水量的变化,脑出血后神经功能评分,ELISA方法测定脑组织内20-HETE含量,Fluoro Jade B染色标记变性坏死的神经元数量(FJB阳性细胞),免疫荧光染色检测钙结合蛋白(Ibal)和抗髓过氧化物酶(MPO)表达等脑组织小胶质细胞和中性粒细胞炎症反应的情况。结果与对照组相比,阻断剂组脑组织内20-羟二十烷四烯酸表达量均明显降低(N=8,p0.01),脑损伤体积明显降低(N=5,P0.05),脑水肿降低(N=8,P0.05),HET0016明显降低脑出血周围组织内FJB阳性细胞数量(N=5,P0.01),降低激活的小胶质细胞和浸润的中性粒细胞的数量(N=5,P0.05)。结论 20-羟二十烷四烯酸在脑出血后加重出血周围神经损伤,其抑制剂可以对脑出血起到治疗作用。