Hui-Lin Cheng

Potential contribution of nuclear factor-κB to cerebral vasospasm after experimental subarachnoid hemorrhage in rabbits

Nuclear factor-κB (NF-κB) plays a key role in inflammation, which is involved in the development of cerebral vasospasm after subarachnoid hemorrhage (SAH). In the present study, we assessed the potential role of NF-κB in regulation of cerebral vasospasm. Nuclear factor-κB DNA-binding activity was measured in cultured vascular smooth muscle cells (VSMCs) treated with hemolysate and pyrrolidine dithiocarbamate (PDTC, 80 μmol/L), an inhibitor of NF-κB. Forty-two rabbits were divided into three groups: control, SAH, and PDTC groups (n = 14 for each group). The caliber of the basilar artery was evaluated. Nuclear factor-κB DNA-binding activity and the gene expression levels of cytokines and adhesion molecules in the basilar artery were measured. Immunohistochemical study was performed to assess the expression and localization of tumor necrosis factor (TNF)-α, intercellular adhesion molecule (ICAM)-1, and myeloperoxidase (MPO). It was observed that NF-κB DNA-binding activity was significantly increased by treatment with hemolysate in cultured VSCMs, but this increase was suppressed by pretreatment with PDTC. Severe vasospasm was observed in the SAH group, which was attenuated in the PDTC group. Subarachnoid hemorrhage could induce increases of NF-κB DNA-binding activity and the gene expression levels of TNF-α, interleukin (IL)-1β, ICAM-1, and vascular cell adhesion molecule (VCAM)-1, which were reduced in the PDTC group. Immunohistochemical study demonstrated that the expression levels of TNF-α, ICAM-1, and MPO were all increased in the SAH group, but these increases were attenuated in the PDTC group. Our results suggest that NF-κB is activated in the arterial wall after SAH, which potentially leads to vasospasm development through induction of inflammatory response.

Intraoperative indocyanine green angiography in intracranial aneurysm surgery: Microsurgical clipping and revascularization.

BACKGROUND The goal of this report is to illustrate the use of intraoperative indocyanine green (ICG) angiography in the surgical management of intracranial aneurysms, including microsurgical clipping and revascularization. METHODS This study included a series of 45 patients who were surgically treated between June 2007 and May 2008 for intracranial aneurysms. Fourty-three of the patients had anterior circulation aneurysms, and 2 had posterior circulation aneurysms. Forty-one patients were treated with microsurgical clipping. Four patients underwent revascularization combined with aneurysm dissection or trapping. Intraoperative ICG angiography was used to visualize the aneurysm clipping, patency of parent artery or graft. The ICG angiography technique is described, with particular reference to evaluation of the aneurysm clipping and revascularization. RESULTS Eighty-nine ICG angiography procedures were performed in 45 patients with intracranial aneurysms. The aneurysms were completely obliterated for all patients, and the grafts were patented for all except 1 patient. Pre-clipping ICG angiography showed the relationship of aneurysm and its parent artery clearly. After aneurysms being clipped, intraoperative ICG angiography found remnant of aneurysms, stenosis or occlusion of parent arteries and grafts in 8 cases, which were revised in the same surgical procedure. The results of ICG angiography correlated well with postoperative DSA in 97% patients. CONCLUSION ICG angiography can provide real-time information and guide revision in the same surgical procedure for the management of intracranial aneurysms.

Expression of Toll-like receptor 4 in the basilar artery after experimental subarachnoid hemorrhage in rabbits: A preliminary study

Inflammation and immunity play a crucial role in the pathogenesis of cerebral vasospasm after subarachnoid hemorrhage (SAH). Recently, a growing body of evidence indicates that Toll-like receptor (TLR) 4 is vital for inflammation and immunity. Therefore, this study aimed to detect the expression of TLR4 in the basilar artery in a rabbit SAH model and to clarify the potential role of TLR4 in cerebral vasospasm. A total of 48 rabbits were randomly divided into four groups: control group; day 3, day 5, and day 7 groups. Day 3, day 5, and day 7 groups were all SAH groups. The animals in day 3, day 5 and day 7 groups were subjected to injection of autologous blood into cisterna magna twice on day 0 and day 2 and were killed on days 3, 5, and 7, respectively. Cross-sectional area of basilar artery was measured and the TLR4 expression was assessed by immunohistochemistry and Western blot analysis. The basilar arteries exhibited vasospasm after SAH and became more severe on day 3 and 5. The elevated expression of TLR4 was detected after SAH and peaked on day 3 and 5. TLR4 is increasingly expressed in a parallel time course to the development of cerebral vasospasm in a rabbit experimental model of SAH.

Therapeutic potential of peroxisome proliferator-activated receptor gamma agonist rosiglitazone in cerebral vasospasm after a rat experimental subarachnoid hemorrhage model

The pathogenesis of cerebral vasospasm is closely associated with inflammation and immune response in arterial walls. Recently, the authors proved the key role of Toll-like receptor (TLR)4 in the development of vasospasm in experimental subarachnoid hemorrhage (SAH) model. Because peroxisome proliferator-activated receptor (PPAR) gamma agonists are identified as effective inhibitors of TLR4 activation, we investigated the anti-inflammation properties of PPAR-gamma agonist rosiglitazone in basilar arteries in a rat experimental SAH model and evaluated the effects of rosiglitazone on vasospasm. Inflammatory responses in basilar arteries were assessed by immunohistochemical staining for intercellular molecule (ICAM)-1 and myeloperoxidase (MPO). Expression of TLR4 was determined by western blot analysis. The degree of cerebral vasospasm was evaluated by measuring the mean diameter and cross-sectional area of basilar arteries. Rosiglitazone suppressed the SAH-induced inflammatory responses in basilar arteries by inhibiting the TLR4 signalling. Furthermore, rosiglitazone could attenuate cerebral vasospasm following SAH. Therefore, we suggested that PPAR-gamma agonists may be potential therapeutic agents for cerebral vasospasm.

Peroxisome proliferator-activated receptor gamma agonist rosiglitazone attenuates oxyhemoglobin-induced Toll-like receptor 4 expression in vascular smooth muscle cells

Inflammation and immune response have been implicated in the pathogenesis of cerebral vasospasm after subarachnoid hemorrhage (SAH). Recently, increased TLR4 expression has been associated with the development of cerebral vasospasm in a rabbit model of SAH. Peroxisome proliferator-activated receptor gamma (PPARgamma) agonists, effective inhibitors of TLR4 activation, may modulate the vasospasm progression via their anti-inflammation effects. We investigate whether the blood component oxyhemoglobin (OxyHb) can induce the expression of Toll-like receptor (TLR) 4 in vascular smooth muscle cells (VSMCs), and evaluate the modulatory effects of PPARgamma agonist rosiglitazone on OxyHb-induced inflammation in VSMCs. Cultured VSMCs incubated with or without rosiglitazone were exposed to OxyHb at 10muM for up to 48h. Expression of TLR4 was assessed by immunocytochemistry and Western blot analysis. Production of tumor necrosis factor alpha (TNF-alpha) in conditioned medium were quantified by ELISA. A marked increase of TLR4 production and TNF-alpha release was observed at 48h after cells were treated with OxyHb. Rosiglitazone reduced TLR4 immunocytochemistry staining and protein production significantly in VSMCs. A specific antagonist for PPARgamma, GW9662, could reverse the anti-inflammatory effects of rosiglitazone. The results demonstrated that OxyHb exposure could induce TLR4 activation in cultured VSMCs. Rosiglitazone suppressed TLR4 expression and cytokine release via the activation of PPARgamma and may have a therapeutic potential for the treatment of vasospasm following SAH.

Epidural cooling for selective brain hypothermia in porcine model

SummaryBackground. Hypothermia has been shown to be neuroprotective in many animal models and several human trials of brain ischemic and trauma. However systemic hypothermia may result in fatal complications. This study was undertaken to test epidural cooling as a new method of inducing selective brain hypothermia.Method. Six adult swine (mean mass, 33.8 ± 3.6 kg) were studied. Anesthesia was maintained with pentobarbital sodium (25 mg kg−1, i.v.) and pancuronium bromide (0.5 mg kg−1 h−1, i.v.). Five probes were placed in the rectum, deep brain, brain surface, epidural space, and room air for temperature monitoring respectively. Epidural cooling was performed using cold-saline (4 °C) perfusion into the epidural space through a flexible double-lumen catheter. The dripping speed of cold saline was controlled to maintain the target temperature. The changes of the epidural space pressure, complete blood counts, basic metabolic panels, tests for fibrinolysis and coagulation status were monitored to assess hypothermia-induced changes.Findings. Following the epidural cooling perfused with cold-saline (4 °C) at mean dripping speed of 720 ml per hour, the local brain surface temperature could decrease rapidly to 17.3–21.6 °C within one minute, and deep brain temperature decreased to 27.2–29.7 °C within 5 minutes. The target temperatures were easily controlled by the dripping speed of cold saline (from mild to deep hypothermia). The rectal temperature was maintained at normal range within 6 perfusion hours. No arrhythmia was observed, and all hematological variables were within the normal range for swine. No increased intracranial pressure was induced by the cooling method.Conclusions. The data demonstrate that epidural space cooling was technically feasible and useful for selective brain hypothermia, and the target temperatures are easily controlled. The induction of hypothermia was rapid and maintained for a long period of time, whereas the body temperature was maintained within the normal range and without hemodynamic instability.

Preliminary study on the effect of trauma-induced secondary cellular hypoxia in brain injury

Secondary cerebral hypoxia has recently been shown to play an important role in the outcome of patients suffering from traumatic brain injury (TBI). However, the precise mechanisms underlying secondary cerebral hypoxia are complex and interrelated. In this study, we investigate the effect of hypoxia within a rat model of trauma-induced late cerebral cortex injury. Using the hypoxia marker pimonidazole, we verified and isolated areas of the cortex that had suffered hypoxic damage. Using subsequent reverse-transcriptase PCR analyses, we found that the expressions of both transforming growth factor beta1 (TGF-beta1) and hypoxia-inducible factor-1alpha (HIF-1alpha) increased significantly under hypoxic conditions induced by TBI compared with uninjured control animals. In addition, the maximum mRNA expression of TGF-beta1 and HIF-1alpha was found at 3 days and 12h after TBI, respectively. Our data suggest that secondary cerebral hypoxia injury involves various cytokines including TGF-beta1 and HIF-1alpha. Furthermore, upon immunohistochemical analysis, both TGF-beta1 and HIF-1alpha expression were almost localized in the same types of cells by using immunohistochemical study. These results may have important implications in the understanding of trauma-induced secondary cerebral hypoxia injury.

Quantitative detection of the expression of mitochondrial cytochrome c oxidase subunits mRNA in the cerebral cortex after experimental traumatic brain injury.

Secondary brain damage plays a critical role in the outcome of patients with traumatic brain injury (TBI). The multiple mechanisms underlying secondary brain damage, including posttraumatic cerebral ischemia, glutamate excitotoxicity, oxidative stress, calcium overload and inflammation, are associated with increased mortality and morbidity after head injury. TBI is documented to have detrimental effects on mitochondria, such as alterations in glucose utilization and the depression of mitochondrial oxidative phosphorylation. Studies on mitochondrial metabolism have provided evidence for dysfunction of the cytochrome oxidase complex of the electron transport chain (complex IV) after TBI. A growing body of evidence indicates that cytochrome c oxidase is vital for mitochondrial oxidative phosphorylation. Therefore, this study aimed to detect the expression of cytochrome c oxidase (CO) mRNA in a rat weight-dropping trauma model and to clarify the differences between injured cortex (IC) and contralateral cortex (CC) after TBI. A total of forty-four rats were randomly assigned to 7 groups: control groups (n=4), sham-operated group (n=20), 6 h, 1 d, 3 d, 5 d and 7 d postinjury groups (n=4 for each group). The group consisted of sham-operated animals underwent parietal craniotomy without TBI. The rats in postinjury groups were subjected to TBI. The rats of control group were executed immediately without TBI or craniotomy after anesthesia. The brain-injured and sham-operated animals were killed on 6 h, 1 d, 3 d, 5 d and 7 d, respectively. Tissue sections from IC and CC were obtained and the expression of cytochrome c oxidase I, II, and III (CO I, II, III) mRNA, three mitochondrial encoded subunits of complex IV, were assessed by Real-time quantitative PCR. A reduction of CO I, II, and III mRNA expression was detected from IC and reduced to the lowest on 3 d. By contrast, the mRNA expression from CC suggested a slight elevation. The differences may indicate the degree of metabolic and physiologic dysfunction. Our results will better define the roles of gene expression and metabolic function in long-term prognosis and outcome after TBI. With a considerable understanding of post-injury mitochondrial dysfunction, therapeutic interventions targeted to the mitochondria may prevent secondary brain damage that leads to long-term cell death and neurobehavioral disability.

Zinc as mediator of ubiquitin conjugation following traumatic brain injury

Previous studies have shown that pathological zinc accumulation and deposition of ubiquitinated protein aggregates are commonly detected in many acute neural injuries, such as trauma, epilepsy and ischemia. However, the underlying mechanisms are poorly understood. Here we assessed the effect of zinc on ubiquitin conjugation and subsequent neurodegeration following traumatic brain injury (TBI). First, we found that scavenging endogenous Zn(2+) reduced trauma-induced ubiquitin conjugation and protected neurons from TBI insults in rat hippocampus. Second, we detected both zinc accumulation and increased ubiquitin conjugated protein following brain trauma in human cortical neurons. Our previous study has shown that zinc can induce ubiquitin conjugation in cultured hippocampal neurons. All these findings indicate that alterations in Zn(2+) homeostasis may impair the protein degradation pathway and ultimately cause neuronal injury following traumatic brain injury.

Cognitive assessment in Chinese patients with aneurysmal subarachnoid hemorrhage

A subgroup of patients who survive aneurysmal subarachnoid hemorrhage (SAH) may have significant cognitive deficits. The aim of the current study was to determine the efficiency of cognitive tests and frequency of cognitive impairments associated with aneurysmal SAH in Chinese. A series of 116 patients with aneurysmal SAH were assessed before surgery. Only 37 patients have completed all tests. The other 79 patients had discontinued because of their clinical conditions, recurring severe headaches, refusing or misunderstanding due to low education. We found that one or more cognitive functions were impaired in 70.3% of the 37 patients, SAH patients were especially impaired in functions that are related to visual reproduction, verbal fluency, and executive functions. The results suggest that the patients have impressive cognitive deficits after aneurysmal SAH. A battery of appropriate cognitive tests should be developed for use by doctors and nurses.