Objective : Gyehyuldeung (GHD) has been widely used in oriental medicine for the treatments of cardiovascular and neurological disorders. Thus, its potential facilitatory activity on axonal regeneration was investigated in the rats. Methods: Sprague-Dawley rats were given crush injury at the sciatic nerve and the changes of axon growth after nerve injury on each nerve injury model were investigated with anti-NF-200 antibody, DiI, GAP-43 protein and Cdc2 protein Results : GHD-mediated enhancement of axonal regeneration after crush injury was measured in both qualitative and quantitative ways by immunofluorescence staining with anti-NF-200 antibody and retrograde tracing of fluorescence dye DiI. GAP-43 protein levels were elevated by GHD treatments in the distal injured sciatic nerve and DRG sensory neurons. The neurite outgrowth of DRG sensory neurons was facilitated by GHD treatment when co-cultured with Schwann cells and astrocytes prepared from injured sciatic nerves and injured spinal cord tissues, respectively. It was observed that Cdc2 protein was up-regulated in co-cultured Schwann cells or astrocytes and Cdc2 protein signals were co-localized to a certain extent with those of phospho-vimentin protein. Conclusions : These results suggest that GHD may play a facilitatory role in axonal regeneration by acting on the injured axons and adjacent non-neuronal cells. The current findings may be useful for the development of therapeutic targets through more specific explorations on molecular interactions between herbal components and endogenous factors.
Nervous system is clinically important, and involved in most disorders directly or indirectly. It could be injury and be a source of symptoms. Injury of central or peripheral nervous system injury may affect that mechanism and interrupt normal function. An understanding of the concepts of axonal transport is important for physical therapist who treat injury of nerves. Three connective tissue layers are the endoneurium, perineurium, epineurium. Each has its own special structural characteristics and functional properties. The blood supply to the nervous system is well equipped in all dynamic and static postures with intrinsic and extrinsic vasculation. After nerve injury, alternations in the ionic compression or pressures within this environment may interfere with blood flow and, consequently conduction and the flow of axoplasm. The cytoskeleton are not static. On the contrary, elements of the cytoskeleton are dynamically regulated and are very likely in continual motion. It permits neural mobility. There are different axonal transport systems within a single axon, of which two main flows have been identified : First, anterograde transport system, Secondly, retrograde transport system. The nervous system adapts lengthening in two basic ways. The one is that the development of tension or increased pressure within the tissues, increased intradural pressure. The other is movements that are gross movement and movement occurring intraneurally between the connective tissues and the neural tissues. In this article, we emphasize the biologic aspects of nervous system that influenced by therapeutic approaches. Although identified scientific information in basic science is utilized at clinic, we would attain the more therapeutic effects and develop the physical therapy science.
Background: Though a number of studies have described the distribution of substance P(SP)-like immunoreactivity in the spinal cord, they have been focused on lamina I and II of the dorsal horn and there are little morphological studies on the topographic distribution and ultrastructure of the SP immunoreactive neurons especially in the ventral horn of the spinal cord. this study was conducted to identify distribution pattern of SP immunoreactive neurons and to difine the synaptic organization of their processes in ventral horn of the thoracic cord of the cat by preembbeding immunocytochemical method using SP antiserum. Methods: Five adults cats of either sex were used and deeply anesthetized by intramuscular injection of ketamine. After removal of the spinal cord, samples of thoracic cord were taken and placed in fresh fixative at $4^{\circ}C$ for 2 hours. Transverse sections $50{\mu}m$ thick were processed using the preembbeding immunocytochemical method and incubated consecutively in the specific primary antibody and the 10% normal goat serum, the rabbit anti-substance P antiserum, the biotin-labelled goat anti-rabbit IgG and finally the avidin-biotin-peroxidase complex. The processed tissue sections were throughly washed and stained in the black with 1% uranyl acetate. Section were examined on a electron microscope. Results: 1) SP immunoreactive neurons were observed in the gray matter around central canal. 2) In lamina I and II SP immunoreactivity was observed in both myelinated and unmyelinated nerve fibers, but in ventral horn only in the unmyelinated nerve fibers. 3) SP immunoreactive axon terminals with small round and large dense core vesicles made chemical synapses onto the dendrites of motor neurons in the ventral horn. Conclusion: SP immunoreactive neurons might play an important role in modulation of motor neurons in the ventral horn of the thoracic cord of the cat.
Ha, Yoon;Yoon, Do Heum;Yeon, Dong Su;Kim, Hyun Ok;Lee, Jin Ju;Cho, Yong Eun;Choi, Joong Uhn
Journal of Korean Neurosurgical Society
/
v.30
no.8
/
pp.963-969
/
2001
Objectives : Cord blood stem cells have been widely used as donor cells for bone marrow transplantation recently. These cells can give rise to a variety of hematopoietic lineages to repopulate the blood. Recent observations reveal that some bone marrow cells and bone marrow stromal cells(MSCs) can grow to become either neurons or glial cells. It is, however, unclear whether or not there exists stems cells which can differentiate into neurons in the blood during the early stages of postnatal life. Methods : Human cord blood stem cells were prepared from human placenta after full term delivery. To induce neuronal differentiation of stem cells, ${\beta}$-mercaptoethanol was treated. To confirm the neuro-glial characteristics of differentiated stem cells, immunocytochemical stain for NeuN, neurofilament, glial fibrillary acidic protein(GFAP), microtubule associated protein2(MAP2) was performed. RT-PCR was performed for detecting nestin mRNA and MAP2 mRNA. Results : We showed in this experiment that neuro-glial markers(NeuN, neurofilament, MAP2, GFAP) were expressed and axon-like cytoplasmic processes are elaborated in the cultured human cord blood stem cells prepared from new born placenta after full term delivery. Nestin mRNA was also detected in fresh cord blood monocytes. Conclusions : These results suggest that human cord blood derived stem cells may be potential sources of neurons in early postnatal life.
Kim Bang-Oul;Kim Jeong-Sang;Kim Kyung-Soo;Jeon Sang-Yoon;Hong Seok
Herbal Formula Science
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v.11
no.2
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pp.173-184
/
2003
Objectives: This study investigates the effects of Salviae miltiorrhizae Radix, Carthami Flos on Brain ischemia of the rats induced from the Occlusion of Lt. Common Carotid Artery. Methods: I observed effects using light microscopes and examined tissue of parietal lobe and hippocampus and VEGF-immunoreactive cells. Results: A small number of VEGF-immunoreactive cells are observed in the control group. VEGF-immunoreactive cells in Salviae miltiorrhizae Radix-administered group were slightly increased compared with control group. VEGF-immunoreactive cells in Carthami Flos-administered group were significantly increased compared with control group. Neurons of parietal lobe and pyramidal cells of hippocampus in the control group were greatly damaged.(neuronal densitity, form of dendrite and axon) On the other hand, neurons of parietal lobe and pyramidal cells of hippocampus in Salviae miltiorrhizae Radix-administered group were less damaged. Neurons of parietal lobe and pyramidal cells of hippocampus in Carthami Flos-administered group were significantly less damaged compared with control group. Conclusion : Salviae miltiorrhizae Radix, Carthami Flos can effect on stimulating angiogenesis and reducinging the damage of neurons in the rats induced from the Occlusion of Lt. Common Carotid Artery.
The present study was performed to observe the ultrastructure of the integument of adult Paragonimus westermani. Dogs experimentally infected with 60 metacercariae of F. westermani were autopsied 4 months after the infection. Adult p. westermani were extracted from the dogs and the fine structure was studied by means of scanning and transmission electron microscope. The findings are as follows: 1. Scanning electron microscopic findings showed that the spines and the papillae are distributed at whole body surface but the well developed spines or papaillae are shown around the oral sucker and ventral sucker. 2. At the end of the body, excretory pore was found, the shape was irregular. 3. Transmission electron microscopic findings showed that plasma membrane, tegument, basal lamina, connective tissue, circular muscle layer, longitudinal muscle layer. nerve axon and tegumental cell were observed. 4. In higher magnification, plasma membrance and bar-shaped granules were found at the outer surface of the tegument.
Objectives : In conditions of brain infarction, irreversible axon damage occurs in the central nerve system (CNS), because gliosis becomes a physical and a mechanical barrier to axonal regeneration. Reactive gliosis induced by ischemic injury such as middle cerebral artery occlusion is involved with up-regulation of GFAP and CD81. This study was undertaken to examine the effect of the Gongjin-dan (GJD) on CD81 and GFAP expression and its pathway in the rat brain following middle cerebral artery occlusion (MCAO). Methods : In order to study ischemic injuries on the brain, infarction was induced by MCAO using insertion of a single nylon thread, through the internal carotid artery, into a middle cerebral artery. Cresyl violet staining, cerebral infarction size measurement, immunohistochemistry and microscopic examination were used to detect the expression of CD81 and GFAP and the effect on the infarct size and pyramidal cell death in the brain of the rat with cerebral infarction induced by MCAO. Also, c-Fos and ERK expression were measured to investigate the signaling pathway after GJD administration in MCAO rats. Results : Measuring the size of cerebral infarction induced by MCAO in the rat after injection of GJD showed the size had decreased. GJD administration showed pyramidal cell death protection in the hippocampus in the MCAO rat. GJD administration decreased GF AP expression in the MCAO rat. GJD administration decreased CD81 expression in the MCAO rat. GJD administration induced up-regulation of c-FOS expression compared with MCAO. GJD administration induced down-regulation of ERK expression compared with MCAO. Conclusion : We observed that GJD could suppress the reactive gliosis, which disturbs the axonal regeneration in the brain of a rat with cerebral infarction after MCAO by controlling the expression of CD81 and GFAP. The effect may be modulated by the regulation of c-Fos and ERK. These results suggest that GJD can be a candidate to regenerate CNS injury.
The development of small granule-containing cell in the superior cervical ganglion was studied by electron microscopic method in human fetuses ranging from 40 mm to 260 mm crown rump length (10 to 30 weeks of gestational age). At 40 mm fetus, the superior cervical ganglion was composed of clusters of undifferentiated cells, primitive neuroblasts, and unmyelinated nerve fibers together with blood vessels. At 90 mm fetus, the superior cervical ganglion consisted of neuroblasts, satellite cell, small granule-containing cells, and unmyelinated nerve fibers. Two morphological types of the small granule-containing cells in the superior cervical ganglion were first indentified at 90 mm fetus, but were rare. Type I granule-containing cell occurred in solitary and had long processes, whereas type II cells tend to appeared in clusters near the blood capillaries. The granule-containing cells were characterized by the presence of dense-cored vesicles ranging from $150{\sim}300nm$ in diameter in both the cell bodies and processes. Other organelles included abundant mitochondria, rough endoplasmic reticulum, neurotubules, and widely distributed ribosomes. The granule-containing cells had long processes similar to those found in principal ganglionic cells. They could be identified by their content in dense-cored vesicles. The small granule-containing cells increased somewhat in size and number with increase of fetal age. Synaptic contacts were first found on the solitary granule-containing cell at 150 mm fetus. Synaptic contacts between the soma and processes of type I granule-containing cells and preganglionic axon terminals were observed. In addition, synaptic junctions between the processes of granule-containing cells and presumed dendrite of postganglionic neuron were also observed from 150 mm onward. On the basis of these features type I granule-containing cells could be considered as interneurons. The clusters of type II granule-containing cells were located in the interstitial or subcapsular portions of the ganglion, and had short processes which ended in close relation to fenestrated capillaries. Therefore it may be infer that clusters of type II granule-containing cells have an endocrine function.
Object: Reactive gliosis that is induced by central nervous system (CNS) injury is involved with up-regulation of CD81 and GFAP. The present study was to examine the effect of the Salvia miltiorrhiza on CD81 and GFAP regulation following brain injury. Methods: Immunoblot and ELISA methods were used to define the level of CD81 and GFAP in the astrocyte cultured from rat brain. Then immunohistochemistry was used to detect CD81 and GFAP in the injured rat brain. Results: The following results were obtained. 1. We did western blot and ELISA to detect the protein isolated from the whole cell and they showed that CD81 and GFAP decreased. 2. We injected Salvia miltiorrhiza extract intravenously to brain-injured rats for 7 days and 30 days, and the immunohistochemistry analyses showed that CD81 and GFAP decreased significantly. Conclusion: These results indicate that Salvia miltiorrhiza could suppress the reactive gliosis, which disturbs the neural regeneration following CNS injury, by controlling the expression of CD81 and GFAP.
Objectives : Effects of Bogijetong-tang (BJT) on peripheral nerve regeneration have been reported in a previous study on BJT but additional study on a damaged peripheral neuropathy where its damage level is physically and chemically more severe was needed. Plus, this study was conducted because there haven't been any studies for BJT on central nerve regeneration. Methods : In order to check the effect on central nerve regeneration, the study on cerebellum cells was started and the sciatic nerve was used to observe the effects on a peripheral nerve which was severely damaged both physically and chemically. Nerve recovery effects were observed by analyzing target proteins such as phospho-extracellular signal-regulated kinase, ${\beta}1$ integrin, neurofilament 200, growth-associated protein-43, cyclin-dependent kinase 1, phospho-vimentin, phospho-Smad, and caspase 3. Results : The significant changes of target protein in cerebellum neurons have been observed. The changes of index protein on the axon regeneration and the nerve recovery in the sciatic nerve have been observed and the effects on cell protection were observed, as well. Conclusions : This study confirmed that BJT made a significant influence on nerve protection and recovery of a damaged peripheral neuropathy and it also made a possibility of its regeneration in a damaged central nerve injury.
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