Cerebral Infarction In Rats Research Articles

Ipsilateral versus bilateral limb-training in promoting the proliferation and differentiation of endogenous neural stem cells following cerebral infarction in rats.

We investigated the effects of ipsilateral versus bilateral limb-training on promotion of endogenous neural stem cells in the peripheral infarct zone and the corresponding cerebral region in the unaffected hemisphere of rats with cerebral infarction. Middle cerebral artery occlusion was induced in Wistar rats. The rat forelimb on the unaffected side was either wrapped up with tape to force the use of the paretic forelimb in rats or not braked to allow bilateral forelimbs to participate in training. Daily training consisted of mesh drum training, balance beam training, and stick rolling training for a total of 40 minutes, once per day. Control rats received no training. At 14 days after functional training, rats receiving bilateral limb-training exhibited milder neurological impairment than that in the ipsilateral limb-training group or the control group. The number of nestin/glial fibrillary acidic protein-positive and nestin/microtubule-associated protein 2-positive cells in the peripheral infarct zone and in the corresponding cerebral region in the unaffected hemisphere was significantly higher in rats receiving bilateral limb-training than in rats receiving ipsilateral limb-training. These data suggest that bilateral limb-training can promote the proliferation and differentiation of endogenous neural stem cells in the bilateral hemispheres after cerebral infarction and accelerate the recovery of neurologic function. In addition, bilateral limb-training produces better therapeutic effects than ipsilateral limb-training.

Effects of secretome obtained from normoxia-preconditioned human mesenchymal stem cells in traumatic brain injury rats

The aim of the present study was to investigate whether secretome obtained from normoxia-preconditioned human mesenchymal stem cells causes attenuation of traumatic brain injury (TBI), induced by fluid percussion injury. Anaesthetized male Sprague-Dawley rats, immediately after the onset of fluid percussion TBI, were divided into two major groups and given the vehicle solution (1 mL) or secretome (500 µg) intravenously. Another group of rats was used as sham-operated controls. The functional outcome such as motor (maximum grasp angle) was evaluated by an inclined plane test. Cellular infarction volume was calculated by triphenyltetrazolium chloride staining. Neuronal loss, apoptosis, and neurotrophic factor such as vascular epithelial growth factor (VEGF) expression in the cortex were measured by immunofluorescence staining. All the parameters were assessed on Day 3 after injury. Compared with those of the sham-operated controls, the motor deficits and cerebral infarction of rats after TBI were significantly attenuated by secretome therapy. The TBI-induced neuronal loss and apoptosis were also significantly reduced by secretome therapy. Furthermore, VEGF-positive cells in the ischemic cortex after TBI were further significantly increased by secretome therapy. Our results demonstrate that intravenous injection of secretome from normoxia-preconditioned human mesenchymal stem cells may ameliorate TBI-induced rats by reducing neuronal cell loss and apoptosis and promoting VEGF production, resulting in functional outcome improvement. We also recommend that secretome from normoxia-preconditioned human mesenchymal stem cells could be a promising treatment strategy for traumatic brain injury.

K-134, a Phosphodiesterase 3 Inhibitor, Prevents Brain Damage by Inhibiting Thrombus Formation in a Rat Cerebral Infarction Model

BackgroundK-134 is a more potent antiplatelet drug with a selective inhibitory effect on phosphodiesterase 3 (PDE3) compared with its analogue, cilostazol.ObjectivesThis study was performed to compare the ameliorating effects of K-134 and cilostazol on brain damage in an experimental photothrombotic cerebral infarction model.Methods and ResultsWe investigated the effects of oral preadministration of PDE3 inhibitors in a rat stroke model established by photothrombotic middle cerebral artery (MCA) occlusion. K-134 significantly prolonged MCA occlusion time at doses >10 mg/kg, and reduced cerebral infarct size at 30 mg/kg in the stroke model (n = 12, 87.5±5.6 vs. 126.8±7.5 mm3, P<0.01), indicating its potent antithrombotic effect. On the other hand, the effects of cilostazol on MCA occlusion time and cerebral infarct size are relatively weak even at the high dosage of 300 mg/kg. Furthermore, K-134 blocked rat platelet aggregation more potently than cilostazol in vitro. Also in an arteriovenous shunt thrombosis model, K-134 showed an antithrombotic effect greater than cilostazol.ConclusionsThese findings suggest that K-134, which has strong antithrombotic activity, is a promising drug for prevention of cerebral infarction associated with platelet hyperaggregability.

Poster 541 The Effect of Swimming Exercise on Cognitive Function in Cerebral Infarction Rats

PM&RVolume 4, Issue 10S p. S375-S376 2012 AAPM&R Annual Assembly Abstract Poster 541 The Effect of Swimming Exercise on Cognitive Function in Cerebral Infarction Rats In-Sung Choi MD, PhD, In-Sung Choi MD, PhD Chonnam National University Medical School & Hospital, Gwangju city, Korea, Republic ofSearch for more papers by this authorJae-Young Han MD, Jae-Young Han MD Chonnam National University Medical School & Hospital, Gwangju city, Korea, Republic ofSearch for more papers by this authorJung-Kook Kim MS, Jung-Kook Kim MS Chonnam National University Medical School & Hospital, Gwangju city, Korea, Republic ofSearch for more papers by this authorSam-Gyu Lee MD, PhD, Sam-Gyu Lee MD, PhD Chonnam National University Medical School & Hospital, Gwangju city, Korea, Republic ofSearch for more papers by this authorMin-Keun Song MD, Min-Keun Song MD Chonnam National University Medical School & Hospital, Gwangju city, Korea, Republic ofSearch for more papers by this author In-Sung Choi MD, PhD, In-Sung Choi MD, PhD Chonnam National University Medical School & Hospital, Gwangju city, Korea, Republic ofSearch for more papers by this authorJae-Young Han MD, Jae-Young Han MD Chonnam National University Medical School & Hospital, Gwangju city, Korea, Republic ofSearch for more papers by this authorJung-Kook Kim MS, Jung-Kook Kim MS Chonnam National University Medical School & Hospital, Gwangju city, Korea, Republic ofSearch for more papers by this authorSam-Gyu Lee MD, PhD, Sam-Gyu Lee MD, PhD Chonnam National University Medical School & Hospital, Gwangju city, Korea, Republic ofSearch for more papers by this authorMin-Keun Song MD, Min-Keun Song MD Chonnam National University Medical School & Hospital, Gwangju city, Korea, Republic ofSearch for more papers by this author First published: 23 October 2012 https://doi.org/10.1016/j.pmrj.2012.09.1141Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat No abstract is available for this article. Volume4, Issue10SOctober 2012Pages S375-S376 RelatedInformation

Poster 542 Comparison of Treadmill and Swimming Exercises on the Cognitive Function in Cerebral Infarction Rats

PM&RVolume 4, Issue 10S p. S186-S186 2012 AAPM&R Annual Assembly Abstract Poster 542 Comparison of Treadmill and Swimming Exercises on the Cognitive Function in Cerebral Infarction Rats In-Sung Choi MD, PhD, In-Sung Choi MD, PhD Chonnam National University Medical School & Hospital, Gwangju city, Korea, Republic ofSearch for more papers by this authorJae-Young Han MD, Jae-Young Han MD Chonnam National University Medical School & Hospital, Gwangju city, Korea, Republic ofSearch for more papers by this authorJung-Kook Kim MS, Jung-Kook Kim MS Chonnam National University Medical School & Hospital, Gwangju city, Korea, Republic ofSearch for more papers by this authorSam-Gyu Lee MD, PhD, Sam-Gyu Lee MD, PhD Chonnam National University Medical School & Hospital, Gwangju city, Korea, Republic ofSearch for more papers by this authorMin-Keun Song MD, Min-Keun Song MD Chonnam National University Medical School & Hospital, Gwangju city, Korea, Republic ofSearch for more papers by this author In-Sung Choi MD, PhD, In-Sung Choi MD, PhD Chonnam National University Medical School & Hospital, Gwangju city, Korea, Republic ofSearch for more papers by this authorJae-Young Han MD, Jae-Young Han MD Chonnam National University Medical School & Hospital, Gwangju city, Korea, Republic ofSearch for more papers by this authorJung-Kook Kim MS, Jung-Kook Kim MS Chonnam National University Medical School & Hospital, Gwangju city, Korea, Republic ofSearch for more papers by this authorSam-Gyu Lee MD, PhD, Sam-Gyu Lee MD, PhD Chonnam National University Medical School & Hospital, Gwangju city, Korea, Republic ofSearch for more papers by this authorMin-Keun Song MD, Min-Keun Song MD Chonnam National University Medical School & Hospital, Gwangju city, Korea, Republic ofSearch for more papers by this author First published: 23 October 2012 https://doi.org/10.1016/j.pmrj.2012.09.624Citations: 1Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat No abstract is available for this article.Citing Literature Volume4, Issue10SOctober 2012Pages S186-S186 RelatedInformation

Artificial Cold Wave-Induced Cerebral Infarction in Rats with Carotid Atherosclerosis

To investigate the effects of cold on blood pressure, serum endothelin-1 content, serum nitric oxide content, and morbidity of cerebral infarction, as well as assess the therapeutic effect of nimodipine. A total of 200 rats were initially assigned to a normal group (n = 10), sham group (n = 10), and carotid atherosclerosis group (n = 180), and subsequently the animals in the carotid atherosclerosis group were randomly assigned to three groups: non-cold (n = 59), cold treatment (n = 58), and nimodipine (n = 58). Rats in the cold and nimodipine groups experienced an artificial cold wave. The temperature was set at 22°C for 12 h (7:00 am to 7:00 pm) and then at 4°C for another 12 h (7:00 pm to 7:00 am), representing a cycle. The animals underwent three cycles of cold. Rats in the nimodipine group were treated with nimodipine and those in the cold group with given an equal volume of intragastric normal saline for 3 days. Hematoxylin and eosin staining showed features of carotid atherosclerosis in all animals. Blood pressure fluctuated with alteration of temperature. A temperature decrease was accompanied by an increase of blood pressure and elevation of serum levels of endothelium-1 and nitric oxide. In addition, although nimodipine could prevent the cold-induced increase of blood pressure and elevation of serum endothelium-1 and nitric oxide levels, it had no effect on blood pressure fluctuation or morbidity of cerebral infarction. The results suggest that dramatic variation in temperature is one of the main causes of cold-induced fluctuation of blood pressure.

Effects of exercise training on synaptic plasticity in rats with focal cerebral infarction

To examine the roles of exercise training in the improvement of damaged neural function and synaptic plasticity. An infarction model was induced by left middle cerebral artery occlusion (MCAO). A total of 70 adult Sprague-Dawley rats were randomly divided into 3 groups: physical exercise group (n = 30) undergoing running wheel exercise daily after MCAO, control group (n = 30) and sham-operated group (n = 10). The latter two groups were fed in standard cages without any special training exercise. The rats were scarified at Days 3, 7, 14, 21 and 35 for the evaluation of neural function by neurological severity scores (NSS). And the synaptic ultrastructures at peri-infarction region were examined by specific marker synaptophysin (SYN). Synaptic ultrastructures at peri-infarction region were observed in both the control and exercise training groups. The presynaptic and postsynaptic membranes were relatively intact. And the presynaptic membranes had more synaptic vesicles from Day 7 post-ischemia. The number of SYN positive cells significantly increased in the exercise training group (21 d: 0.8 ± 0.1; 35 d: 0.7 ± 0.1) versus those in the control group (21 d: 0.4 ± 0.1; 35 d: 0.5 ± 0.1) at Days 21 and 35 post-ischemia (P < 0.05). Moreover, the neurological severity scores in the exercise training group (7 d: 7.8 ± 0.8; 14 d: 5.6 ± 0.8; 21 d: 3.3 ± 0.8; 35 d: 3.0 ± 0.8) showed a quicker declination versus those in the control group (7 d: 8.8 ± 0.7; 14 d: 7.7 ± 0.9; 21 d: 6.9 ± 0.8; 35 d: 4.2 ± 0.8) from Day 7 post-ischemia (P < 0.05). Exercise training plays an important role in the recovery of damaged neural function and synaptic plasticity after cerebral infarction in rats.

Influence of acupuncture on neural movement function in rats with middle cerebral artery occlusion-A randomized controlled trial

ObjectivesTo observe recovery in movement function in rats with middle cerebral artery occlusion (MCAO) after acupuncture treatment. MethodsAccording to the randomized and controlled principle 1384 rats were divided into the basic control group (including the normal, sham, model control, model without intervention, Nimodipine, and para-Renzhong groups) and the acupuncture group (including the Neiguan (PC 6), Weizhong (BL 40), Sanyinjiao (SP 6), Chize (LU 5), Renzhong (GV 6) and non-acupoint groups). MCAO was modeled by Zea-longa's thread ligation and rats with scores of 1–3, as assessed by Zausinger's six-point method, were used in this study. Moreover, in the acupuncture group each acupoint was set with 12 different parameters by the orthogonal intersection method, resulting in 78 groups with 18 rats per group. The rats were treated by acupuncture once every 12 h for a total of six sessions and neurobehavioral scores were measured after each session. The neurobehavioral scores were compared by one-way ANOVA using the statistical software SPSS 17.0. ResultsAfter acupuncture therapy the mean neurobehavioral scores in MCAO rats increased gradually at each time point with a significant difference among the six scores, but with no significant differences between the fourth (48 h) and the fifth score (60 h), and between the fifth (60 h) and the sixth (72 h) score (P> 0.05). ConclusionsMCAO rats gradually recovered movement function over multiple acupuncture sessions. After the fouth acupuncture session (48 h), the neurobehavioral scores of rats with cerebral infarction remained stable. Acupuncture treatment had a reliable curative effect on movement function in cerebral infarction rats.

Abstract 117: Therapeutic Impact of Direct Cell Sheet Transplantation Derived from Bone Marrow Stromal Cells for Cerebral Infarct in Rats

Background and Purpose - Less-invasive and efficient donor cell delivery is quite important for the clinical application of cell transplantation therapy. Recently, cell sheet technology has gained a great interest as a novel tissue engineering technology, but there are no studies that evaluate the validity of the bone marrow stromal cell (BMSC) sheets as the donor for cerebral infarct. The aim of this study is to assess whether the BMSC sheet can be an alternative transplantation technique to promote functional recovery after cerebral infarct. Methods - The BMSC were harvested from green fluorescence protein (GFP)-transgenic rat and were cultured on a temperature-responsive culture dishes. The mono-layered BMSC sheet was detached by changing the temperature of culture dishes, and was histologically analyzed. Next, the SD rats were subjected to permanent middle cerebral artery occlusion and were divided into 3 experimental groups. Thus, the vehicle or BMSC suspension was stereotactically transplanted into the ipsilateral striatum, or the BMSC sheet was directly transplanted onto the ipsilateral intact neocortex at 7 days after the insult (n=9, 9, 7, respectively). Motor function was assessed for 28 days after transplantation. Finally, histological analysis was performed. Results - BMSC sheet was composed of 9.8 ± 2.4 x10 5 cells (n=10). Stereotactic injection of BMSC significantly enhanced the recovery of motor function after cerebral infarct. Likewise, non-invasive cell sheet transplantation also significantly promoted functional recovery. The therapeutic effect was comparable to stereotactic cell transplantation. Fluorescence immunohistochemistry revealed that the GFP-positive cells were densely distributed in the dorsal neocortex adjacent to cerebral infarct in both groups. Double fluorescence immunohistochemistry demonstrated that the majority of engrafted GFP-positive cells were positive for NeuN and morphologically simulated the neurons in both groups. The density of reactive astrocytes in the ipsilateral striatum was less pronounced in cell sheet transplantation group than in stereotactic transplantation group. Conclusion - These findings strongly suggest that cell sheet technology would make it possible to non-invasively deliver the BMSC into the infarct brain and to yield significant therapeutic effects, when compared with stereotactic injection of cell suspension. The technology may be a clinically valuable and less-invasive construct for non-invasive and efficient cell delivery to regenerate the infarct brain.

A subpopulation of endothelial progenitor cells with low aldehyde dehydrogenase activity attenuates acute ischemic brain injury in rats

Previous studies have examined the therapeutic effect of endothelial progenitor cells (EPCs) during the chronic phase of cerebral infarction in rats; however, few studies have investigated the effects of EPCs during the acute phase of infarction. In this study, we evaluated the therapeutic effect of EPCs with low aldehyde dehydrogenase activity (Alde-Low EPCs) in rats with acute cerebral infarction, and our results provide insight that may help to identify a therapeutic mechanism of EPCs for acute cerebral infarction. The administration of Alde-Low EPCs into rats with acute cerebral infarction results in the accumulation and migration of the Alde-Low EPCs into the infarct area and the subsequent decrease of infarct volume. Moreover, we found that the stromal cell-derived factor-1 (SDF-1) and CXC chemokine receptor 4 (CXCR4) signaling pathway may regulate the accumulation of Alde-Low EPCs. The transplantation of Alde-Low EPCs may represent a potential treatment strategy for acute cerebral infarction.

The Effect of Combined Therapy of Exercise and Nootropic Agent on Cognitive Function in Focal Cerebral Infarction Rat Model

ObjectiveTo investigate the effect of combined therapy of exercise and nootropic agent on cognitive function in a focal cerebral infarction rat model.MethodForty 10-week old male Sprague-Dawley rats were subjected to photothrombotic cerebral infarction of the left parietal lobe. All rats were randomly divided into 4 groups: group A was photothrombotic cerebral infarction rats without any treatment (n=10); group B was photothrombotic cerebral infarction rats with swimming exercise (n=10); group C was photothrombotic cerebral infarction rats with oral administration of acetyl-L-carnitine (n=10); group D was photothrombotic cerebral infarction rats with swimming exercise and oral administration of acetyl-L-carnitine (n=10). Cognitive function was evaluated using the Morris water maze test on the 1st day, and the 1st, 2nd, and 4th week after the induction of cerebral infarction. The activity of superoxide dismutase (SOD) and the level of malondialdehyde (MDA) in the hippocampus were measured. The neuronal cells of the hippocampus were histopathologically evaluated.ResultsThe escape latency was shorter in groups B, C, and D than in group A. However, the differences were not statistically significant at the 1st, 2nd and 4th week. The activity of SOD was the highest in group D. The level of MDA was the lowest in group D. We observed more normal neuronal cells in groups B, C, and D.ConclusionThe combined therapy of exercise and nootropic agent was helpful in ameliorating oxidative stress in the focal cerebral infarction rat model. However, the effect did not translate into improvement of cognitive function.

Abstract 8694: Phosphodiesterase 3 Inhibitor Prevents Thrombotic Diseases Without Affecting Hemostatic Function

Introduction: Conventional anti-platelet therapy is associated with increased risk of bleeding. Recently, the results of the second Cilostazol Stroke Prevention Study (CSPS 2) showed similar efficacy but fewer hemorrhagic events with the phosphodiesterase (PDE)3 inhibitor cilostazol vs. aspirin. Hypothesis: We hypothesized that PDE3 inhibition reduces thrombus formation without affecting hemostasis, and leads to beneficial effects in experimental cerebral infarction and peripheral artery disease (PAD) models Methods: We used a cilostazol analog, K-134, as a more potent and selective PDE3 inhibitor. To evaluate the anti-thrombus activity of K-134, we measured mortality rate in a pulmonary embolism model established by collagen infusion into mice via the tail vein 10 min after oral administration of K-134. We also studied its influence on tail vein bleeding times. To investigate whether K-134 can ameliorate other thrombotic diseases, we established a rat cerebral infarction model by photochemically induced thrombosis-mediated occlusion of middle cerebral arteries and a PAD model by laurate injection-mediated peripheral arterial injury. Pathological signs in the areas of necrosis in the hind legs in the PAD model were classified as Grade 0 to 5. Results: K-134 decreased mortality in the mouse pulmonary embolism model when orally administered at 3, 10, and 30 mg/kg ( n = 29-30, P &lt; 0.01 vs. control, ED50 = 2.1 mg/kg) but did not prolong bleeding time even at 100 mg/kg compared to control ( n = 30, 98 ± 6 vs. 84 ± 7 s., not significant). In contrast, warfarin prolonged bleeding time ( n = 29-30, 139 ± 7 vs. 84 ± 7 s., P &lt; 0.01). Preoperative administration of K-134 at 30 mg/kg reduced brain infarct size compared to control in the rat cerebral infarction model ( n = 12, 87.5 ± 5.6 vs. 126.8 ± 7.5 mm 3 , P &lt; 0.01). Furthermore, 30 mg/kg K-134 inhibited the progression of lesions compared with controls in the PAD model ( n = 12-15, mean grade of pathological sign: 2.3 ± 0.2 vs. 3.5 ± 0.3, P &lt; 0.05). Conclusion: PDE3 inhibition is an attractive approach for the secondary prevention of cerebral infarction and treatment of peripheral arterial disease.

Intracerebral, but not intravenous, transplantation of bone marrow stromal cells enhances functional recovery in rat cerebral infarct: An optical imaging study

Recent studies have indicated that bone marrow stromal cells (BMSC) may improve neurological function when transplanted into an animal model of CNS disorders, including cerebral infarct. However, there are few studies that evaluate the therapeutic benefits of intracerebral and intravenous BMSC transplantation for cerebral infarct. This study was aimed to clarify the favorable route of cell delivery for cerebral infarct in rats. The rats were subjected to permanent middle cerebral artery occlusion. The BMSC were labeled with near infrared (NIR)-emitting quantum dots and were transplanted stereotactically (1 × 10⁶ cells) or intravenously (3 × 10⁶ cells) at 7 days after the insult. Using in vivo NIR fluorescence imaging technique, the behaviors of BMSC were serially visualized during 4 weeks after transplantation. Motor function was also assessed. Immunohistochemistry was performed to evaluate the fate of the engrafted BMSC. Intracerebral, but not intravenous, transplantation of BMSC significantly enhanced functional recovery. In vivo NIR fluorescence imaging could clearly visualize their migration toward the cerebral infarct during 4 weeks after transplantation in the intracerebral group, but not in the intravenous, group. The BMSC were widely distributed in the ischemic brain and some of them expressed neural cell markers in the intracerebral group, but not in the intravenous group. These findings strongly suggest that intravenous administration of BMSC has limited effectiveness at clinically relevant timing and intracerebral administration should be chosen for patients with ischemic stroke, although further studies would be warranted to establish the treatment protocol.

Validity of Bone Marrow Stromal Cell Expansion by Animal Serum-Free Medium for Cell Transplantation Therapy of Cerebral Infarct in Rats—A Serial MRI Study

This study was aimed to test the hypothesis that human bone marrow stromal cells (hBMSC) expanded in fetal calf serum (FCS)-free, platelet lysate (PL)-containing medium would retain their capacity of migration, survival, and neural differentiation when transplanted into the infarct brain, using serial in vivo magnetic resonance imaging (MRI). Cell growth kinetic analysis revealed that hBMSC maintain their proliferative activity when cultured either in conventional FCS-containing medium or FCS-free, PL-containing medium. Subsequently, hBMSC were labeled with a superparamagnetic iron oxide agent and were stereotactically transplanted into the ipsilateral striatum of rats at 7days after permanent middle cerebral artery occlusion. Serial MRI performed over 8weeks revealed that they retain their migratory capacity towards the cerebral infarct. Moreover, double fluorescence immunohistochemistry also revealed that they preserve their capacity of differentiation into the neural cells in the peri-infarct area. The hBMSC expanded in the FCS-free, PL-containing medium retain their capacity of migration, survival, and differentiation when stereotactically transplanted into the infarct brain. The present findings strongly suggest the clinical utility of PL as a substitute to expand autologous hBMSC for cerebral infarct in the future.

Effects of acupuncture in combination with exercise on learning, memory and the expression of microtubuleassociated protein-2 in the hippocampal CA3 region after experimental focal cerebral infarction in rats

Objective To study the expression of microtubule-associated protein-2 (MAP-2) in the CA3 region of the hippocampus and any effect of acupuncture combined with exercise training on the recovery of learning and memory after experimental cerebral infarction.Methods Eighty Wistar rats were randomly divided into a sham-operation group (n = 8 ) and an operation group (n = 72 ) subdivided randomly and equally into a model group, an exercisetraining group and an acupuncture plus exercise training group, with 24 rats in each group. After right middle cerebral artery occlusion or a mock operation, the expression of MAP-2 in the CA3 region of the hippocampus was detected with immunohistochemistry 1, 3 and 5 weeks after the operation. Learning and memory function was assessed at the 5th week post operation.Results MAP-2- positive fibers were arranged in an orderly way and distributed densely in the shamoperation group. After cerebral infarction, number of MAP-2 positive neurons and dendrite fibers decreased in the CA3 region of the affected side. A week after the operation, MAP-2 positive fibers had increased significantly in the acupuncture plus exercise group compared with the model group and the exercise group. At the 3rd and 5th week post operation, expression of MAP-2 and optical density had increased significantly compared with the model group and the exercise training group. At the 5th week post operation, Y maze discrimination and learning tests showed that learning and memory had improved significantly more in the acupuncture plus exercise training group compared with the model group and the exercise training group.Conclusions Exercise training combined with acupuncture can promote changes in dendritic plasticity and increase MAP-2 expression in the CA3 region after cerebral infarction. This may be correlated positively with the recovery of learning and memory after cerebral infarction, at least in rats. Key words: Cerebral infarction; Learning; Memory; Acupuncture; Exercise training; Microtubule-associated protein-2 ; Hippocampus

Therapeutic Impact of Human Bone Marrow Stromal Cells Expanded by Animal Serum–Free Medium for Cerebral Infarct in Rats

The donor cell culture in animal serum-free medium is important for the clinical application of cell transplantation therapy. Recently, human-derived platelet lysate (PL) gained interest as a substitute for fetal calf serum (FCS), but there are no studies that evaluate the validity of human bone marrow stromal cells (hBMSCs) expanded with PL-containing medium for central nervous system disorders. To test the hypothesis that hBMSCs expanded with FCS-free, PL-containing medium can promote functional recovery after cerebral infarct. hBMSCs were cultured in the FCS- or PL-containing medium. Cell-growth kinetics were analyzed. The vehicle or hBMSCs was stereotactically transplanted into the ipsilateral striatum of the rats subjected to permanent middle cerebral artery occlusion 7 days after the insult. Motor function was assessed for 8 weeks, and the fate of transplanted hBMSCs was examined using immunohistochemistry. There was no significant difference in hBMSC expansion between the 2 groups. Transplantation of hBMSCs expanded with the FCS- or PL-containing medium equally promoted functional recovery compared with the vehicle group. Histological analysis revealed that there were no significant differences in their migration, survival, and neural differentiation in the infarct brain between the 2 groups. hBMSCs expanded with PL-containing medium retained their capacity of migration, survival, and differentiation and significantly promoted functional recovery when stereotactically transplanted into the infarct brain. The PL may be a clinically valuable and safe substitute for FCS in expanding hBMSCs to regenerate the infarct brain.

Is the GIRK Channel a Possible Target in the Development of a Novel Therapeutic Drug of Urinary Disturbance?

Clinically, both overactive bladder (OAB) and dysuria are known to occur in patients with cerebral infarction (CI). A few anticholinergic drugs are used to treat OAB in such patients, although the effect is not satisfactory. On the other hand, little or no therapeutic drug is available for dysuria after CI. We previously reported that dextromethorphan (DM) and cloperastine (CP), centrally acting antitussives, reduce the frequency of micturition reflex and increase the threshold pressure in anesthetized rats. In this article, we describe the effects of DM and CP on urinary disturbances at 24 h after CI, induced by occlusion of the left middle cerebral artery in conscious rats. We also briefly review the structure, function, and distribution of G-protein-coupled inwardly rectifying K(+) (GIRK) channels in the brain, since both drugs have potent inhibitory effect on GIRK channel-activated currents in brain neurons. Of the two drugs, CP at antitussive-effective doses ameliorated both OAB and dysuria 24 h after CI in rats. On the other hand, DM aggravated the dysuria, although it significantly ameliorated the OAB. These results suggest that CP may have some therapeutic value for the treatment of OAB and dysuria after CI. At the present time, mechanisms of the effect of CP are unknown. However, several lines of evidence including pharmacological findings support the idea that the effects of CP may be produced at least partly by an increase in the level of 5-HT in the brain through an inhibitory effect on GIRK channel-activating currents.

Volatile anesthetics as a neuroprotective agent

Pharmacologic protection of brain from ischemic insult has been intensively investigated since the early 1960s [1] and a considerable amount of work has been performed to show the effectiveness of various drugs. Among these studies, anesthetic agents, including intravenous and volatile drugs, were shown to be promising agents. In an article published in this issue of the Korean Journal of Anesthesiology, Park et al. [2] present the neuroprotective effect of sevoflurane on forebrain ischemic injury in a rat model. They show the protective effect of sevoflurane in near-complete brain ischemia and the influence of time on the ischemia in the rat brain. In previous studies, the effects of anesthetic agents on ischemic insults were shown to vary depending on the ischemic intensity. Among these studies, Miura et al. [3] demonstrated that isoflurane, ketamine or nitrous oxide and fentanyl had neuroprotective effects against incomplete brain ischemic, but in near-complete global ischemic, only isoflurane was effective. In addition, Park et al. confirmed that in near-complete ischemia, sevoflurane also displayed neuroprotective properties in rats. Volatile anesthetics were shown to be neuroprotective in an acute ischemic rat model. Park et al. performed TUNEL staining, which measures apoptosis of neurons, on the 7th day after ischemic insult and did not observe any differences between the control and sevoflurane group. However, when using an incomplete brain ischemia model, Pape et al. [4] found that sevoflurane had long-term effects on neuronal cell damage and expression of apoptotic factors. Engelhard et al. [5] also investigated the effect of propofol on neuronal ischemia after incomplete cerebral ischemia. They found reduced neuronal cell damage and favorable modulation of apoptosis-regulating proteins over long-term period. But Bayona et al. [6] showed that propofol infusion was protective until 3 days after ischemic insult; however, after 3 weeks there were no differences compared to the control group. Kawaguchi et al. [7] concluded that isoflurane delays but does not prevent cerebral infarction in rats subjected to focal ischemia. In this regard, it seems that the long-term neuroprotective action of anesthetic agents is only effective when the ischemic insult is mild and the neuroprotective effect is not sustained for near-complete ischemic. But these shortterm effects in near-complete ischemia can still save time to allow for the use of other agents directed against apoptosis. Park et al. also demonstrate the effect of ischemic duration on the severity of ischemia. It is reasonable to conclude that shorter duration produces less damage than longer duration. These combined finding indicate that volatile anesthetics are promising agents to prevent neuronal injury during the early ischemic period and provide additional time for the administration of other neuroprotective agents.

The effects of early exercise on brain damage and recovery after focal cerebral infarction in rats

AimExercise can be used to enhance neuroplasticity and facilitate motor recovery after a stroke in rats. We investigated whether treadmill running could reduce brain damage and enhance the expression of midkine (MK) and nerve growth factor (NGF), increase angiogenesis and decrease the expression of caspase-3.MethodsSeventy-seven Wistar rats were split into three experimental groups (ischaemia-control: 36, ischaemia-exercise: 36, sham-exercise: 5). Stroke was induced by 90-min left middle cerebral artery occlusion using an intraluminal filament. Beginning on the following day, the rats were made to run on a treadmill for 20 min once a day for a maximum of 28 consecutive days. Functional recovery after ischaemia was assessed using the beam-walking test and a neurological evaluation scale in all rats. Infarct volume, and the expression of MK, NGF, anti-platelet-endothelial cell adhesion molecule (PECAM-1), and caspase-3 were evaluated at 1, 3, 5, 7, 14 and 28 days after the induction of ischaemia.ResultsOver time motor coordination and neurological deficits improved more in the exercised group than in the non-exercised group. The infarct volume in the exercised group (12.4 ± 0.8%) subjected to treadmill running for 28 days was significantly decreased compared with that in the control group (19.8 ± 4.2%, P < 0.01). The cellular expression levels of MK, NGF and PECAM-1 were significantly increased while that of caspase-3 was decreased in the peri-infarct area of the exercised rats.ConclusionsOur findings show that treadmill exercise improves motor behaviour and reduces neurological deficits and infarct volume, suggesting that it may aid recovery from central nervous system injury.

Effects of electric stimulation on motor function and the expression of Rho kinase following cerebral infarction

Objective To investigate the effects of electric stimulation on motor function and expression of Rho kinase following cerebral infarction in rats. Methods Acute cerebral infarction was modeled in adult male Sprague-Dawley ( SD) rats using the permanent middle cerebral artery occlusion ( PMCAO) technique. The rats were randomly divided into sham operation, control, unilateral electric stimulation and bilateral electric stimulation groups ( each group had 36 rats). Electric stimulation was applied to the paralyzed ( unilateral or bilateral) limbs in the last two groups. Motor function recovery and the expression of Rho kinase were examined using a beam walking test ( BWT) and immunohistochemistry respectively at the 3rd, 7th, 14th and 21st day after stimulation. In addition, the cerebral infarction volume was also determined by 2, 3, 5-triphenyl tetrazolium chloride (TTC) staining at different time points. Results Motor function improved significantly in the electric stimulation groups compared with the control group, and the effect in the bilateral electric stimulation group was better than that in the unilateral electric stimulation group. The expression of Rho kinase decreased remarkably in the electric stimulation groups, and it was significantly lower in the bilateral group than in the unilateral electric stimulation group. No difference in cerebral infarction volume was found at the 3rd day. At the 21st day, the cerebral infarction volume had decreased significantly in both stimulation groups compared with the control group, but no difference was seen between the unilateral and bilateral electric stimulation groups. Conclusions Early electric stimulation, especially bilateral electric stimulation, can improve motor function after cerebral infarction and reduce cerebral infarction volume, which may be associated with down-regulation of the expression of Rho kinase in the border zone of the infarction. Key words: Electric stimulation; Cerebral infarction; Motor function; Rho kinase