高压氧对急性脑创伤后皮层一氧化氮合酶mRNA表达的影响

目的:拟观察高压氧(HBO)治疗对急性创伤性颅脑损伤后皮层NOSmRNA表达的影响,探讨HBO治疗急性脑损伤的机理。方法:采用自由落体法打击模型制备SD大鼠急性脑创伤,伤后1 h、12 h采用0.25 MPaHBO治疗,伤后6 h、24 h取样皮层,应用半定量逆转录聚合酶链反应(RT-PCR)观察神经元型一氧化氮合酶(nNOS)、内皮型一氧化氮合酶(eNOS)和诱导型一氧化氮合酶(iNOS)mRNA表达量变化。结果:0.25MPaHBO治疗各时间组nNOS、eNOS和iNOSmRNA较急性颅脑损伤各时间组显著下降(P<0.01),且HBO治疗24 h组较6 h组下降更明显(P<0.05,P<0.01),0.25 MPa常氧高氮各时间组与急性颅脑损伤各时间组NOSmRNA表达量无统计学意义。结论:HBO治疗可以下调nNOSmRNA、iNOSmRNA和eNOSmRNA的表达量,可能为HBO治疗脑创伤的机理之一。     

AMD3100对局灶脑缺血/再灌注大鼠大脑皮质血管再生的影响

目的探讨AMD3100阻断SDF-1/CXCR4轴后,对局灶脑缺血/再灌注大鼠缺血半暗带血管再生的影响。方法将SD大鼠随机分为假手术组(S组)、模型组(IR组)、AMD3100组(IRA组)、生理盐水组(IRN组)。采用线栓法制备大鼠局灶脑缺血/再灌注模型,缺血2h后将IR、IRA和IRN组分为再灌注12h,1、3和7d四个亚组。HE染色观察局灶脑缺血/再灌注后大脑皮质病理变化。免疫组化法检测CD31在缺血半暗带表达。荧光定量PCR检测外周血中AC133mRNA表达。结果与IRN组比较,IRA 12h外周血中AC133mRNA显著升高,第1d升高达峰值(P0.01),IRA 3dAC133mRNA表达比IRA1d显著减少(P0.05);与IRN组比较,IRA组CD31阳性血管密度在第1d无显著变化(P0.05),第3和7d血管密度显著减少(P0.01);IRA 7d梗死区由大量坏死神经细胞和泡沫细胞填充,坏死较严重。结论持续注射AMD3100能动员干/祖细胞快速进入外周血,但可能抑制局灶脑缺血/再灌注大鼠缺血半暗带血管再生,加重梗死区坏死。     

Immunocytochemical studies of astrocytes following injury to the cerebral cortex of the rat

The morphological change of cerebral cortex astrocytes from protoplasmic to glial fibrillary acidic protein (GFAP)-containing cells is induced by injury. Protoplasmic astrocytes that contain no detectable amount of GFAP become filled with GFAP and their processes extend to form the glial scar around the wound. It is hypothesized that this transformation is induced by cAMP and neurotransmitters released from damaged neuronal cells. A similar mechanism may be present in other brain regions following injury or disease.     

The expression changes of Numblike in rat brain cortex after traumatic brain injury

Numblike (Numbl) plays an important role in ependymal wall integrity and subventricular zone neuroblast survival. And Numbl is specifically expressed in the brain. However, its expression and function in the central nervous system lesion are still unclear. In this study, we performed a traumatic brain injury (TBI) model in adult rats and investigated the dynamic changes of Numbl expression in the brain cortex. Western blot and immunohistochemistry analysis revealed that Numbl was present in normal brain. It gradually decreased, reached the lowest point at day 3 after TBI, and then increased during the following days. Double immunofluorescence staining showed that Numbl immunoreactivity was found in neurons, but not astrocytes and microglia. Moreover, the 3rd day post injury was the apoptotic peak implied by the alteration of caspase-3. All these results suggested that Numbl may be involved in the pathophysiology of TBI and further research is needed to have a good understanding of its function and mechanism.     

Increased expression of BAG-1 in rat brain cortex after traumatic brain injury

BAG-1 protein was initially identified as a Bcl-2-binding protein. It was reported to enhance Bcl-2 protection from cell death, suggesting that BAG-1 represents a new type of anti-cell death gene. Moreover, recent study has shown that BAG-1 can enhance the proliferation of neuronal precursor cells, attenuate the growth inhibition induced by siah1. However, its function and expression in the central nervous system lesion are not been understood very well. In this study, we performed a traumatic brain injury (TBI) model in adult rats and investigated the dynamic changes of BAG-1 expression in the brain cortex. Double immunofluorescence staining revealed that BAG-1 was co-expressed with NEURON and glial fibrillary acidic protein (GFAP). In addition, we detected that proliferating cell nuclear antigen had the co-localization with GFAP, and BAG-1. All our findings suggested that BAG-1 might involve in the pathophysiology of brain after TBI.     

Upregulation of p21-activated Kinase 6 in rat brain cortex after traumatic brain injury

p21-activated Kinase 6 (PAK6) is a serine/threonine kinase belonging to the p21-activated kinase (PAK) family. PAK kinases are well-known regulators of a wide variety of cellular functions, including regulation of cytoskeleton rearrangement, cell survival, apoptosis and the mitogen-activated protein kinase signaling pathway. To elucidate the expressions and possible functions of PAK6 in central nervous system (CNS) lesion and repair, we performed a traumatic brain injury (TBI) model in adult rats. Western blot analysis revealed that PAK6 level significantly increased at day 3 after damage, and then declined during the following days. Besides, double immunofluorescence staining showed PAK6 was primarily expressed in the neurons and a few of glial cells in the normal group. While after injury, the expression of PAK6 was increased significantly in the astrocytes and neurons, and the astrocytes had largely proliferated. We also examined the expression of proliferating cell nuclear antigen (PCNA) whose change was correlated with the expression of PAK6. Importantly, double immunofluorescence staining revealed that cell proliferation evaluated by PCNA appeared in many PAK6-expressing cells at day 3 after injury. In addition, injury-induced expression of PAK6 was co-labeled by active caspase-3 during neuronal apoptosis after injury. Collectively, we hypothesized PAK6 may play important roles in CNS pathophysiology after TBI and further research is needed to have a good understanding of its function and mechanism.     

Simulation of angiogenesis in three dimensions: Application to cerebral cortex

The vasculature is a dynamic structure, growing and regressing in response to embryonic development, growth, changing physiological demands, wound healing, tumor growth and other stimuli. At the microvascular level, network geometry is not predetermined, but emerges as a result of biological responses of each vessel to the stimuli that it receives. These responses may be summarized as angiogenesis, remodeling and pruning. Previous theoretical simulations have shown how two-dimensional vascular patterns generated by these processes in the mesentery are consistent with experimental observations. During early development of the brain, a mesh-like network of vessels is formed on the surface of the cerebral cortex. This network then forms branches into the cortex, forming a three-dimensional network throughout its thickness. Here, a theoretical model is presented for this process, based on known or hypothesized vascular response mechanisms together with experimentally obtained information on the structure and hemodynamics of the mouse cerebral cortex. According to this model, essential components of the system include sensing of oxygen levels in the midrange of partial pressures and conducted responses in vessel walls that propagate information about metabolic needs of the tissue to upstream segments of the network. The model provides insights into the effects of deficits in vascular response mechanisms, and can be used to generate physiologically realistic microvascular network structures.     

大脑皮层缺血损伤后CSF-1R在中枢神经元的表达变化

目的：观察大脑皮层缺血损伤后,集落刺激因子-1受体（CSF-1R）在损伤区及其周围脑区表达的变化,探讨集落刺激因子-1（CSF-1）/CSF-1R信号在中枢神经系统缺血损伤、修复过程中的作用。方法：采用免疫组化技术和Western blot法,对正常和受缺血损伤刺激小鼠全脑CSF-1R免疫阳性神经细胞的分布及表达量进行对比观察。结果：缺血损伤能引起CSF-1R在中枢神经系统（CNS）广泛脑区神经元上的表达上调,不仅发生在损伤及其周围区,还出现在损伤对侧及许多远隔部位。结论：CSF-1/CSF-1R可能参与CNS缺血损伤的发生和修复过程。     

Activation of P2X7 promotes cerebral edema and neurological injury after traumatic brain injury in mice

Traumatic brain injury (TBI) is a leading cause of death and disability worldwide. Cerebral edema, the abnormal accumulation of fluid within the brain parenchyma, contributes to elevated intracranial pressure (ICP) and is a common life-threatening neurological complication following TBI. Unfortunately, neurosurgical approaches to alleviate increased ICP remain controversial and medical therapies are lacking due in part to the absence of viable drug targets. In the present study, genetic inhibition (P2X7-/- mice) of the purinergic P2x7 receptor attenuated the expression of the pro-inflammatory cytokine, interleukin-1β (IL-1β) and reduced cerebral edema following controlled cortical impact, as compared to wild-type mice. Similarly, brilliant blue G (BBG), a clinically non-toxic P2X7 inhibitor, inhibited IL-1β expression, limited edemic development, and improved neurobehavioral outcomes after TBI. The beneficial effects of BBG followed either prophylactic administration via the drinking water for one week prior to injury or via an intravenous bolus administration up to four hours after TBI, suggesting a clinically-implementable therapeutic window. Notably, P2X7 localized within astrocytic end feet and administration of BBG decreased the expression of glial fibrillary acidic protein (GFAP), a reactive astrocyte marker, and attenuated the expression of aquaporin-4 (AQP4), an astrocytic water channel that promotes cellular edema. Together, these data implicate P2X7 as a novel therapeutic target to prevent secondary neurological injury after TBI, a finding that warrants further investigation.     

Alterations of cerebral cortex and hippocampal proteasome subunit expression and function in a traumatic brain injury rat model

Following cellular stress or tissue injury, the proteasome plays a critical role in protein degradation and signal transduction. The present study examined the β-subunit expression of constitutive proteasomes (β1, β2, and β5), immunoproteasomes (β1i, β2i, and β5i) and the 11S proteasome activator, PA28α, in the rat CNS after traumatic brain injury (TBI). Concomitant measures assessed changes in proteasome activities. Quantitative real time PCR results indicated that β1 and β2 mRNA levels were not changed, while β5 mRNA levels were significantly decreased in injured CNS following TBI. However, β1i, β2i, β5i, and PA28α mRNA levels were significantly increased in the injured CNS. Western blotting studies found that β1, β2, β5, β2i, and β5i subunit protein levels remained unchanged in the injured CNS, but β1i and PA28α protein levels were significantly elevated in ipsilateral cerebral cortex and hippocampus. Proteasome activity assays found that peptidyl glutamyl peptide hydrolase-like and chymotrypsin-like activity were significantly reduced in the CNS after TBI, and that trypsin-like proteasome activity was increased in the injured cerebral cortex. Our results demonstrated that both proteasome composition and function in the CNS were affected by trauma. Treatments that preserve proteasome function following CNS injury may be beneficial as an approach to cerebral neuroprotection.     

Bone canaliculus endings in the area of the osteocyte lacuna. Electron-microscopic studies.

Light- and electron-microscopic studies were carried out on the bone canaliculi and their endings in the wall fo the osteocyte lacuna. Two types of canalicular endings were distinguished in the lacuna wall. Those of the first type, fairly numerous, are the endings of small diameter, which do not branch off in the immediate proximity of the lacuna. The other type, occurring in twos or threes, are large in diameter and branch off in the close vicinity of the lacuna, invariably possessing two or three processes which run up to the point of branching of the canaliculus. The images obtained made it possible to demonstrate that the collagen fibrils in both types of endings in the lacuna wall are, in principle, arranged in a similar manner. Nonethless, the type of end formation found where the fibrils run along the axis of the lacuna differs from that for the fibrils in a plexus.     

Correlation of the destructive and reparative processes in neurons of the cerebral cortex in rats after thermal injury

Rat brain sensorimotor cortical neurons were investigated in thermal trauma, using electron microscopic radioautography. RNA synthesis and destructive neuronal changes (chromatolysis, nuclear and mitochondrial damage) have been determined. It has been established that the development of a reparative process--ribosomal RNA synthesis--was roughly parallel to the appearance of destructive changes and was marked in neurons with considerable ultrastructural disturbances.