• Korean Journal of Medicinal Crop Science
• /
• v.12 no.5
• /
• pp.415-423
• /
• 2004

Myristica fragrans seed from Myristica fragrans Houtt (Myristicaceae) has various pharmacological activities peripherally and centrally. The present study aims to investigate the effect of the methanol extract of Myristica fragrans seed (MF) on kainic acid (KA)-induced neurotoxicity in primary cultured rat cerebellar granule neuron. MF, over a concentration range of 0.05 to $5\;{\mu}g/ml$ inhibited KA $(500\;{\mu}M)-induced$ neuronal cell death, which was measured by trypan blue exclusion test and 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) assay. MF $(0.5\;{mu}g/ml)$ inhibited glutamate release into medium induced by KA $(500\;{\mu}M)$, which was measured by HPLC. Pretreatment of MF $(0.5\;{mu}g/ml)$ inhibited KA $(500\;{\mu}M)-induced$ elevation of cytosolic calcium concentration $([Ca^{2+}]_c)$, which was measured by a fluorescent dye, Fura 2-AM, and generation of reactive oxygen species (ROS). These results suggest that MF prevents KA-induced neuronal cell damage in vitro.

• Advances in pediatric surgery
• /
• v.11 no.1
• /
• pp.9-18
• /
• 2005

Abnormal distribution of enteric nerves such as adrenergic, cholinergic and non-adrenergic non-cholinergic nerves (NANC) may cause the failure of relaxation at the involved bowel segment in Hirschsprung's disease (HD). Nitric oxide (NO) is a major inhibitory NANC neurotransmitter in the gastrointestinal tract. NO is synthesized by activation of nNOS (neuronal nitric oxide synthase) in the intramural ganglion cells and regulates bowel movement. To assess the distribution of nNOS in HD, immunohistochemical staining to nNOS was utilized on paraffin embedded specimens. Ten control colon specimens were tested for feasibility of staining. Immunohistochemisrty was done on ganglionic colon as well as aganglionic segment of 15 patients with HD. nNOS immunoreactivity was observed in the neuronal cells, small cells and nerve fibers in the muscle layer and submucosal neuronal cells of control specimens. This finding was also observed in the ganglionic segments of HD. But, there was no nNOS immunoreactivity in aganglionic segments of HD. In conclusion nNOS immunohistochemical staining of paraffin embedded specimen is feasible and reliable. And the results suggest that the relaxation failure of the aganglionic bowel in HD is related to the absence of nNOS containing cells and nerve fibers.

• The Korea Journal of Herbology
• /
• v.27 no.3
• /
• pp.89-94
• /
• 2012

Objects : The purpose of this study was to observe the effect of Salviae Miltriorrhiza Radix(SMR) water-extract on intracerebral hemorrhage(ICH) and neuronal apoptosis in the injured areas. Method : ICH was induced by the stereotaxic intrastriatal injection of bacterial collagenase type IV in Sprague-Dawley rats. The rats were givened oral SMR treatment once a day for three days after the ICH treatment. TUNEL positive cells in the affected regions were performed by TUNEL assay, Bax and Bcl-2 positive cells by immunohistochemistry and the Bax expression by western blotting method. Results : The results are as follow; 1. SMR significantly reduced the number of TUNEL positive cells in the peri-hematoma reigions of ICH-induced rats. 2. SMR significantly reduced the number of Bax positive cells in the peri-hematoma regions of ICH-induced rats. 3. SMR did not affect the number of Bcl-2 positive cells in the peri-hematoma regions of ICH-induced rats. 4. SMR significantly reduced the Bax expressions compared with ICH group in hemorrhagic hemisphere of ICH-induced rats. Conclusion : These results suggest that SMR is effective in reducing neuronal apoptosis.

• Natural Product Sciences
• /
• v.18 no.1
• /
• pp.26-31
• /
• 2012

Actinidia arguta (Actinidiaceae) has been reported to have several pharmacological effects such as anti-inflammatory, anti-allergic, and anti-oxidant activities. The present study investigated the protective activity of an ethanol extract from the leaf and stem of A. arguta against glutamate-induced neurotoxicity using cultured rat cortical neurons. Exposure of cultured cortical neurons to $500{\mu}M$ glutamate for 12 h triggered neuronal cell death. A. arguta inhibited glutamate-induced neuronal death and apoptosis, which were measured by a 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) assay and Hoechst 33342 staining, respectively. The increase of pro-apoptotic proteins, Bax and c-caspase-3, in glutamate-treated neurons was significantly inhibited by treatment with A. arguta. A. arguta also inhibited $500{\mu}M$ glutamate-induced elevation of intracellular calcium concentration ($[Ca^{2+}]_i$) and reactive oxygen species (ROS) generation, which were measured by fluorescent dyes, Fluo-4 AM and $H_2DCF$-DA, respectively. These results suggest that A. arguta may prevent glutamate-induced apoptotic neuronal death by inhibiting $[Ca^{2+}]_i$ elevation and ROS generation and, therefore, may have a therapeutic role for the prevention of neurodegeneration in cerebral ischemic diseases.

• Journal of Microbiology and Biotechnology
• /
• v.19 no.11
• /
• pp.1490-1495
• /
• 2009

The development of nanotechnology has penetrated the fields of biology and medicine, resulting in remarkable applications for tissue regeneration. In order to apply this technology to tissue engineering, we have developed nano-scaled 3D scaffolds consisting of growth factor-loaded heparin/poly(l-lysine) nanoparticles (NPs) attached to the surface of polymeric micro spheres via polyionic complex methods. Growth factor-loaded NPs were simply produced as polyelectrolyte complexes with diameters of 100-200 nm. They were then coated onto positively charged poly(lactic-co-glycolic acid) (PLGA) pretreated with polyethyleneimine to enable cell adhesion, proliferation, and stimulation of neurite outgrowth. Propidium iodide staining and $\beta$-tubulin analysis revealed that neuronal PC12 cells proliferated extensively, expressed significant amounts of b-tubulin, and showed well-structured neurite outgrowth on polymeric microspheres by stimulation with growth factors. These results suggest that cellular adhesion and biological functionality on prepared PLGA microspheres enabled terminal differentiation of neuronal cells.

• Proceedings of the Korean Society of Applied Pharmacology
• /
• 1993.11a
• /
• pp.28-33
• /
• 1993

Regulation of nerve growth factor (NGF)-induced neuronal differentiation by GTPase activating protein(GAP) and its mechanism were investigated in rat pheochromocytoma cell line, PCl2. Overexpression of GAP caused the delay in the onset of neurite outgrowth of PCl2 eel Is in response to NGF. GAP has been known to inhibit p21$\^$ras/, the activated form of which induces neuronal differentiation. Therefore, the activity of p21$\^$ras/ was compared in control cells and cells overexpressing GAP indirectly by measuring the activities of B-Raf and MAP kinase that are known to be positively regulated by p21$\^$ras/. Surprisingly, NGF-induced activities of these two proteins were the same in control eells and GAP-overexpressing cells. Activities of Trk, PLC-r and SMC that act at a site upstream to p21$\^$ras/ in NGF signal transduction pathway were not also affected by GAP overexpression. Interestingly, however, the extent of tyrosine phosphorylation of SNT was found to be remarkably low in cells overexpressing GAP. It has been shown previously that neurotrophins and not mitogens induce SNT tyrosine phosphorylation in PCl2 cells. Thus it is possible that the timing of NGF-induced neuronal differntiation may be in part regulated by SNT and the slower onset of neurite outgrowth in cells overexpressing GAP may be through the inhibition of SNT by GAP.

• Korean Journal of Pharmacognosy
• /
• v.40 no.1
• /
• pp.41-45
• /
• 2009

Parkinson's disease (PD) is the second most common neurodegenerative disorder after Alzhemier's disease. Neuropathologically, PD is characterized by the selective loss of dopaminergic neurons. The neuronal toxicity of cytosolic excess dopamine (DA) has been described in many studies using several cell lines. In dopaminergic neurons, cytosolic excess DA is easily oxidized via monoamine oxidase (MAO)-B, tyrosinase or by auto-oxidation to produce neurotoxic metabolites such as DA quinone. So, in the present study, we induced cell death by treatment of DA ($600{\mu}M$) in human neuroblastoma SH-SY5Y cell which was treated samples before 24 hr, and cell viability was measured by fluorescence activated cell sorter (FACs) analysis. Of those tested, the extracts of Poria cocos (赤茯笭)(whole), Gastrodia elata (rhizomes), Eucommia ulmoides (炒)(barks), Syneilesis palmata (whole), Acorus gramineus (rhizomes), Ligustrum japonicum (leaves) showed neuroprotective effects in dose dependent manner.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.19 no.6
• /
• pp.1546-1551
• /
• 2005

This Study was designed to investigate the effects of Nelumbinis Rhizomatis Nodus (NRN) on the change of cerebral hemodynamics [regional cerebral blood flow (rCBF), pial arterial diameter (PAD) and mean arterial blood pressure (MABP)] in normal and cerebral ischemic rats. And, this study was designed to investigate the inhibition of lactate dehydrogenase activity in neuronal cells The results were as follows NRN significantly increased rCBF and PAD in a dose-dependent manner, and NRN increased MABP in a dose-dependent manner. This results suggested that NRN significantly increased rCBF by dilating PAD. Both rCBF and PAD were significantly and stably increased by NRN (10 mg/kg, i.p.) during the period of cerebral reperfusion, which contrasted with the findings of rapid and marked increase in control group. NRN significantly inhibited lactate dehydrogenase activity in neuronal cells. This results suggested that NRN prevented the neuronal death. It is suggested that NRN had an anti-ischemic effect through the improvement of cerebral hemodynamics and inhibitive effect on the brain damage.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.19 no.6
• /
• pp.1640-1646
• /
• 2005

An oriental medicinal drugs Jahageo (JHG, Hominis placenta) were examined to determine its effects on the responsiveness of central nervous system neurons after injury. We found that JHG was involved in neurite outgrowth of DRG sensory axons. JHG treatment also increased expression of axonal growth-associated protein GAP-43 in DRG sensory neurons after sciatic nerve injury and in the injured spinal cord. JHG treatment during the spinal cord injury increased induction levels of cell division cycle 2 (Cdc2) protein in DRG as well as in the spinal cord. Histochemical investigation showed that induced Cdc2 in the injured spinal cord was found in non-neuronal cells. These results suggest that JHG regulates activities of non-neuronal cells such as oligodendrocyte and astrocyte in responses to spinal cord injury and protects neuronal responsiveness after axonal damage.

• Applied Microscopy
• /
• v.25 no.1
• /
• pp.65-74
• /
• 1995

An immunocytochemical study of GABA-positive neuronal elements was performed at the electron microscopic level to examine subcellular distribution of the inhibitory neurotransmitter in the dog basilar pons. Electron-dense reaction product was observed in neuronal somata and dendritic processes. One or more unlabeled axon terminals made asymmetric synaptic contacts with these GABAergic somatic and dendritic profiles. A large number of GABA-positive axon terminals were also observed. They made symmetric as well as asymmetric synaptic contacts with unlabeled dendritic profiles. In axo-axonic synapses, postsynaptic axon-like processes were consistently GABA-immunoreactive. These observations suggest that the inhibitory local circuit neurons in the dog basilar pons play a major role in cerebro-ponto-cerebellar circuitry by integrating various afferent inputs and conveying them into the cerebellar cortex and the deep cerebellar nuclei.