To investigate the role of AQP9 in brain edema, the expression of AQP9 in an infectious rat brain edema model induced by the injection of lipopolysaccharide (LPS) was examined. Immunohistochemistry and reverse transcription-polymerase chain reaction (RT-PCR) analysis demonstrated that the expressions of AQP9 mRNA and protein at all observed intervals were significantly increased in LPS-treated animals in comparison with the control animals. Time-course analysis showed that the first signs of blood-brain barrier disruption and the increase of brain water content in LPS-treated animals were evident 6 h after LPS injection, with maximum value appearing at 12 h, which coincided with the expression profiles of AQP9 mRNA and protein in LPS-treated animals. The further correlation analysis revealed strong positive correlations among the brain water content, the disruption of the blood-brain barrier and the enhanced expressions of AQP9 mRNA and protein in LPS-treated animals. These results suggested that the regulation of AQP9 expression may play important roles in water movement and in brain metabolic homeostasis associated with the pathophysiology of brain edema induced by LPS injection.
Aquaporins/genetics ; Aquaporins/*metabolism ; Blood-Brain Barrier/metabolism ; Brain/drug effects ; Brain/physiology ; Brain Edema/chemically induced ; Brain Edema/*metabolism ; Lipopolysaccharides ; Rats, Sprague-Dawley ; Water/physiology

BACKGROUNDAlthough some studies have reported that aquaporin-4 (AQP4) plays an important role in the brain edema after traumatic brain injury (TBI), little is known about the AQP4 expression in the early stage of TBI, or about the correlation between the structural damage to the blood-brain barrier (BBB) and angioedema. The aim of this project was to investigate the relationship between AQP4 expression and damage to the BBB at early stages of TBI.

METHODSOne hundred and twenty healthy adult Wistar rats were randomly divided into two groups: sham operation group (SO) and TBI group. The TBI group was divided into five sub-groups according to the different time intervals: 1, 3, 6, 12, and 24 hours. The brains of the animals were taken out at different time points after TBI to measure brain water content. The cerebral edema and BBB changes in structure were examined with an optical microscopy (OM) and transmission electron microscopy (TEM), and the IgG content and AQP4 protein expression in traumatic brain tissue were determined by means of immunohistochemistry and Western blotting. The data were analyzed with SPSS 13.0 statistical software.

RESULTSIn the SO group, tissue was negative for IgG, and there were no abnormalities in brain water content or AQP4 expression. In the TBI group, brain water content significantly increased at 6 hours and peaked at 24 hours following injury. IgG expression significantly increased from 1 to 6 hours following injury, and remained at a high level at 24 hours. Pathological observation revealed BBB damage at 1 hour following injury. Angioedema appeared at 1 hour, was gradually aggravated, and became obvious at 6 hours. Intracellular edema occurred at 3 hours, with the presence of large glial cell bodies and mitochondrial swelling. These phenomena were aggravated with time and became obvious at 12 hours. In addition, microglial proliferation was visible at 24 hours. AQP4 protein expression were reduced at 1 hour, lowest at 6 hours, and began to increase at 12 hours, showing a V-shaped curve.

CONCLUSIONSThe angioedema characterized by BBB damage was the primary type of early traumatic brain edema. It was followed by mixed cerebral edema that consisted of angioedema and cellular edema and was aggravated with time. AQP4 expression was down-regulated during the angioedema attack, but AQP4 expression was upregulated during intracellular edema.

Animals ; Aquaporin 4 ; metabolism ; Blood-Brain Barrier ; metabolism ; Blotting, Western ; Brain Edema ; metabolism ; Brain Injuries ; metabolism ; Immunohistochemistry ; Rats ; Rats, Wistar

OBJECTIVE: To observe the expression of aquaporin 4 (AQP4) in diffuse brain injury (DBI) of rats and to explore the corresponding effect of AQP4 for brain edema. METHODS: The rat model of DBI was established using Marmarou's impact-compression trauma model. Brain water content was measured by dry-wet weight method. Blood-brain barrier permeability was evaluated by Evans blue (EB) staining. Immunohistochemical method was used to observe the expression of AQP4. RESULTS: Brain water content increased after 3 h and peaked at 24 h after DBI. Brain EB content significantly increased and peaked at 12 h after DBI. The expression of AQP4 significantly increased after 3 h and peaked at 24 h after DBI, and the number of AQP4 positive astrocytes increased. CONCLUSION The increment of the permeability of blood-brain barrier and the expression of AQP4 may contribute to the development of brain edema in rat DBI. The change of AQP4 expression in astrocytes may also contribute to determine DBI.
Animals ; Aquaporin 4/metabolism* ; Astrocytes ; Blood-Brain Barrier/metabolism* ; Brain ; Brain Edema/metabolism* ; Brain Injuries/metabolism* ; Cell Membrane Permeability/genetics* ; Disease Models, Animal ; Permeability ; Rats ; Water

Water balance is one of the basic regulation mechanisms of homeostasis. There are 13 subtypes of aquaporins in mammals (AQP0-AQP12). In neural system, the AQP4 is mainly distributed in astrocytes. Phosphorylation and expression regulation of AQP4 is involved in the formation of brain edema, particularly in the clearance of vasogenic edema and the formation of cytotoxic edema. This article reviews regulations and functions of AQP4 in vasogenic edema and cytotoxic edema.
Animals ; Aquaporin 4 ; metabolism ; physiology ; Brain ; metabolism ; physiopathology ; Brain Edema ; metabolism ; physiopathology ; Homeostasis ; Humans ; Water-Electrolyte Balance ; physiology

Brain edema leading to an expansion of brain volume has a crucial impact on morbidity and mortality following traumatic brain injury as it increases intracranial pressure, impairs cerebral perfusion and oxygenation, and contributes to additional ischemic injuries. Classically, two major types of traumatic brain edema exist: "vasogenic" and "cytotoxic/cellular". However, the cellular and molecular mechanisms contributing to the development/resolution of traumatic brain edema are poorly understood and no effective drugs can be used now. Aquaporin-4 (AQP4) is a water-channel protein expressed strongly in the brain, predominantly in astrocyte foot processes at the borders between the brain parenchyma and major fluid compartments, including cerebrospinal fluid and blood. This distribution suggests that AQP4 controls water fluxes into and out of the brain parenchyma. In cytotoxic edema, AQP4 deletion slows the rate of water entry into brain, whereas in vasogenic edema, AQP4 deletion reduces the rate of water outflow from brain parenchyma. AQP4 has been proposed as a novel drug target in brain edema. These findings suggest that modulation of AQP4 expression or function may be beneficial in traumatic brain edema.
Animals ; Aquaporin 4 ; analysis ; antagonists & inhibitors ; chemistry ; physiology ; Brain ; metabolism ; Brain Edema ; drug therapy ; etiology ; Brain Injuries ; complications ; Humans ; Mice

OBJECTIVETo observe the effects of ginseng total saponin (GTS) on the water content in the brain tissue, the activity of superoxide dismutase (SOD), the content of malondialdehyde (MDA), the expression levels of tumor necrosis factor alpha (TNF-alpha) and interleukin 1beta (IL-1beta), and the neurological function in rats with traumatic brain injury (TBI), and to explore the roles of GTS in treating traumatic brain edema rats and its possible mechanisms.

METHODSThe TBI rat model was established using modified Feeney's method. Rats were randomly divided into 3 groups, i.e., the sham-operation group, the TBI group, and the GTS-treated group. All rats were sacrificed after their neurological behavior was scored at day 1, 3, 5, and 7 of TBI. The brain tissue was taken out to measure the brain water content with wet-dry weight method. The activity of SOD in the brain tissue and the content of MDA were determined using biochemistry method. The expression levels of TNF-alpha and IL-1beta in the brain tissue were detected using ELISA.

RESULTSCompared with the TBI group at the same time point, the brain water content and the content of MDA decreased, the activity of SOD increased, the expression levels of TNF-alpha and IL-1beta obviously decreased, and the neurological functions were obviously improved in the GTS-treated group (P<0.05).

CONCLUSIONSGTS could obviously alleviate the degree of traumatic brain edema after TBI, and attenuate the deleted neurological behavioral symptoms. The underlying mechanisms might be achieved through reducing the production of MDA, decreasing the expression levels of TNF-alpha and IL-1beta, elevating the activity of SOD, inhibiting free radical reaction, and alleviating inflammatory reactions.

Animals ; Brain Edema ; metabolism ; Brain Injuries ; metabolism ; Interleukin-1beta ; metabolism ; Male ; Malondialdehyde ; metabolism ; Panax ; Rats ; Rats, Sprague-Dawley ; Saponins ; pharmacology ; Superoxide Dismutase ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

Activated protein C (APC), a natural anticoagulant, has been reported to exert direct vasculoprotective, neural protective, anti-inflammatory, and proneurogenic activities in the central nervous system. This study was aimed to explore the neuroprotective effects and potential mechanisms of APC on the neurovascular unit of neonatal rats with intrauterine infection-induced white matter injury. Intraperitoneal injection of 300 μg/kg lipopolysaccharide (LPS) was administered consecutively to pregnant Sprague-Dawley rats at embryonic days 19 and 20 to establish the rat model of intrauterine infection- induced white matter injury. Control rats were injected with an equivalent amount of sterile saline on the same time. APC at the dosage of 0.2 mg/kg was intraperitoneally injected to neonatal rats immediately after birth. Brain tissues were collected at postnatal day 7 and stained with hematoxylin and eosin (H&E). Immunohistochemistry was used to evaluate myelin basic protein (MBP) expression in the periventricular white matter region. Blood-brain barrier (BBB) permeability and brain water content were measured using Evens Blue dye and wet/dry weight method. Double immunofluorescence staining and real-time quantitative PCR were performed to detect microglial activation and the expression of protease activated receptor 1 (PAR1). Typical pathological changes of white matter injury were observed in rat brains exposed to LPS, and MBP expression in the periventricular region was significantly decreased. BBB was disrupted and the brain water content was increased. Microglia were largely activated and the mRNA and protein levels of PAR1 were elevated. APC administration ameliorated the pathological lesions of the white matter and increased MBP expression. BBB permeability and brain water content were reduced. Microglia activation was inhibited and the PAR1 mRNA and protein expression levels were both down-regulated. Our results suggested that APC exerted neuroprotective effects on multiple components of the neurovascular unit in neonatal rats with intrauterine infection- induced white matter injury, and the underlying mechanisms might involve decreased expression of PAR1.
Animals ; Animals, Newborn ; Blood-Brain Barrier ; Brain Edema ; metabolism ; Cerebrovascular Circulation ; Female ; Male ; Protein C ; metabolism ; Rats ; Rats, Sprague-Dawley

Brachial plexus avulsion (BPA) is a combined injury involving the central and peripheral nervous systems. Patients with BPA often experience severe neuropathic pain (NP) in the affected limb. NP is insensitive to the existing treatments, which makes it a challenge to researchers and clinicians. Accumulated evidence shows that a BPA-induced pain state is often accompanied by sympathetic nervous dysfunction, which suggests that the excitation state of the sympathetic nervous system is correlated with the existence of NP. However, the mechanism of how somatosensory neural crosstalk with the sympathetic nerve at the peripheral level remains unclear. In this study, through using a novel BPA C7 root avulsion mouse model, we found that the expression of BDNF and its receptor TrκB in the DRGs of the BPA mice increased, and the markers of sympathetic nervous system activity including α1 and α2 adrenergic receptors (α1-AR and α2-AR) also increased after BPA. The phenomenon of superexcitation of the sympathetic nervous system, including hypothermia and edema of the affected extremity, was also observed in BPA mice by using CatWalk gait analysis, an infrared thermometer, and an edema evaluation. Genetic knockdown of BDNF in DRGs not only reversed the mechanical allodynia but also alleviated the hypothermia and edema of the affected extremity in BPA mice. Further, intraperitoneal injection of adrenergic receptor inhibitors decreased neuronal excitability in patch clamp recording and reversed the mechanical allodynia of BPA mice. In another branch experiment, we also found the elevated expression of BDNF, TrκB, TH, α1-AR, and α2-AR in DRG tissues from BPA patients compared with normal human DRGs through western blot and immunohistochemistry. Our results revealed that peripheral BDNF is a key molecule in the regulation of somatosensory-sympathetic coupling in BPA-induced NP. This study also opens a novel analgesic target (BDNF) in the treatment of this pain with fewer complications, which has great potential for clinical transformation.
Humans ; Mice ; Animals ; Hyperalgesia/metabolism* ; Brain-Derived Neurotrophic Factor/metabolism* ; Hypothermia/metabolism* ; Neuralgia ; Brachial Plexus/injuries* ; Edema/metabolism*

Animals ; Aquaporin 4 ; metabolism ; Blood-Brain Barrier ; physiopathology ; Brain Edema ; physiopathology ; Brain Ischemia ; metabolism ; physiopathology ; Capillary Permeability ; physiology ; Male ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; metabolism ; physiopathology

The purpose of this study is to investigate the effect of methylprednisolone(M.P.) on the alterations of ATP, sum of adenosine nucleotides, adenylate energy charge(E.C.), glucose and lactate in the cats with acute focal ischemic cerebral edema. The acute occlusion of left middle cerebral artery(MCA) of forty cats for 1,3 and 5 hours respectively were accomplished by applying Heifetz clip through the transorbital approach operating microscope. Twelve cats were not recirculated as a untreated group, twelve cats were recirculated for 2 hours as a ecirculation group and twelve cats were recirculated for 2 hours and given M.P.(15mg/kg) at 30 minutes after occlusion initially, and then every one and a half hour as a treatment group. The experimental results are as follows. 1) In 1-hour untreated group, ATP was reduced to 34.0%, sum of adenosine nucleotides reduced to 72.2%, adenylate E.C. reduced to 60.6%, glucose reduced to 67.3% and lactate increased to 156.6% of the control value. In the recirculation group, ATP was reduced to 42.0%, sum of adenosine nucleotides reduced to 82.4%, adenylate E.C. reduced to 74.3%, glucose increased to 552.7% and lactate decreased to 79.8%. In the treatment group, ATP was increased to 143.9%, sum of adenosine nucleotides increased to 153.9%, adenylate E.C. decreased to 92.9%, glucose increased to 3334.5% and lactate decreased to 74.6%. 2) In 3-hour untreated group, ATP was decreased to 24.9%, sum of adenosine nucleotides reduced to 22.9%, adenylate E.C. reduced to 58.6%, glucose decreased to 45.5% and lactate increased to 161.3% of the control value. In the recirculation group, ATP reduced to 32.9%, sum of adenosine nucleotides reduced to 28.6%, adenylate E.C. reduced to 71.4%, glucose rose to 520.0% and lactate to 135.3% of the control value. In the treatment group, ATP reduced to 99.5%, sum of adenosine nucleotides increased to 103.5%, adenylate E.C. decreased to 84.3%, glucose rose to 1187.3% and lactate increased to 101.2%. 3) In 5-hour untreated group, ATP decreased to 5.3%, sum of adenosine nucleotides reduced to 9.0%, adenylate E.C. reduced to 58.6%, glucose decreased to 25.5% and lactate increased to 187.9%. In the recirculation group, ATP decreased to 4.4%, sum of adenosine nucleotides decreased to 5.8%, adenylate E.C. decreased to 57.1%. In the treatment group, ATP was reduced to 11.2%, sum of adenosine nucleotides and adenylate E.C. reduced to 70.0%, glucose rose to 103.6% and lactate to 157.2% of the control value. As the results shown above, the therapeutic beneficial effects of M.P. were observed in cats of 1 or 3-hour occlusion of MCA with 2-hour recirculation.
Adenosine ; Adenosine Triphosphate ; Animals ; Brain Edema ; Cats ; Energy Metabolism* ; Glucose ; Ischemia* ; Lactic Acid ; Methylprednisolone* ; Nucleotides