• Journal of Ginseng Research
• /
• v.37 no.4
• /
• pp.401-412
• /
• 2013

Korean Red Ginseng (KRG) is an oriental herbal preparation obtained from Panax ginseng Meyer (Araliaceae). To expand our understanding of the action of KRG on central nervous system (CNS) function, we examined the effects of KRG on tissue plasminogen activator (tPA)/plasminogen activator inhibitor-1 (PAI-1) expression in rat primary astrocytes. KRG extract was treated in cultured rat primary astrocytes and neuron in a concentration range of 0.1 to 1.0 mg/mL and the expression of functional tPA/PAI-1 was examined by casein zymography, Western blot and reverse transcription-polymerase chain reaction. KRG extracts increased PAI-1 expression in rat primary astrocytes in a concentration dependent manner (0.1 to 1.0 mg/mL) without affecting the expression of tPA itself. Treatment of 1.0 mg/mL KRG increased PAI-1 protein expression in rat primary astrocytes to $319.3{\pm}65.9%$ as compared with control. The increased PAI-1 expression mediated the overall decrease in tPA activity in rat primary astrocytes. Due to the lack of PAI-1 expression in neuron, KRG did not affect tPA activity in neuron. KRG treatment induced a concentration dependent activation of PI3K, p38, ERK1/2, and JNK in rat primary astrocytes and treatment of PI3K or MAPK inhibitors such as LY294002, U0126, SB203580, and SP600125 (10 ${\mu}M$ each), significantly inhibited 1.0 mg/mL KRG-induced expression of PAI-1 and down-regulation of tPA activity in rat primary astrocytes. Furthermore, compound K but not other ginsenosides such as Rb1 and Rg1 induced PAI-1 expression. KRG-induced up-regulation of PAI-1 in astrocytes may play important role in the regulation of overall tPA activity in brain, which might underlie some of the beneficial effects of KRG on CNS such as neuroprotection in ischemia and brain damaging condition as well as prevention or recovery from addiction.

• Journal of Life Science
• /
• v.28 no.12
• /
• pp.1397-1405
• /
• 2018

Mitochondria have multiple functions in cells: providing chemical energy, storing cellular $Ca^{2+}$, generating reactive oxygen species, and regulating apoptosis. Through these functions, mitochondria are also involved in the maintenance, proliferation, and differentiation of stem/progenitor cells. In the brain, the subventricular zone (SVZ) is one of the neurogenic regions that contains neural stem cells (NSCs) throughout a lifetime. However, reports on the role of mitochondria in SVZ NSCs are scarce. Here, we show that rotenone, a complex I inhibitor of mitochondria, inhibits the proliferation and differentiation of SVZ NSCs in different ways. In proliferating NSCs, rotenone decreases mitosis as measured through phosphorylated histone H3 detection; moreover, apoptosis is not induced by rotenone at 50 nM. In differentiating NSCs, rotenone blocks neurogenesis and oligodendrogenesis while glial fibrillary acidic protein-positive astrocytes are not affected. Interestingly, in this study there were more cells in the differentiating NSCs treated with rotenone for 4-6 days than in the vehicle control group which was a different effect from the reduced number of cells in the proliferating NSCs. We examined both apoptosis and mitosis and found that rotenone decreased apoptosis as detected by staining cleaved caspase-3 but did not affect mitosis. Our results suggest that functional mitochondria are necessary in both the proliferation and differentiation of SVZ NSCs. Furthermore, mitochondria might be involved in the mitosis and apoptosis that occur during those processes.

• Journal of Ginseng Research
• /
• v.45 no.3
• /
• pp.390-400
• /
• 2021

Background: We recently showed that gintonin, an active ginseng ingredient, exhibits antibrain neurodegenerative disease effects including multiple target mechanisms such as antioxidative stress and antiinflammation via the lysophosphatidic acid (LPA) receptors. Amyotrophic lateral sclerosis (ALS) is a spinal disease characterized by neurodegenerative changes in motor neurons with subsequent skeletal muscle paralysis and death. However, pathophysiological mechanisms of ALS are still elusive, and therapeutic drugs have not yet been developed. We investigate the putative alleviating effects of gintonin in ALS. Methods: The G93A-SOD1 transgenic mouse ALS model was used. Gintonin (50 or 100 mg/kg/day, p.o.) administration started from week seven. We performed histological analyses, immunoblot assays, and behavioral tests. Results: Gintonin extended mouse survival and relieved motor dysfunctions. Histological analyses of spinal cords revealed that gintonin increased the survival of motor neurons, expression of brain-derived neurotrophic factors, choline acetyltransferase, NeuN, and Nissl bodies compared with the vehicle control. Gintonin attenuated elevated spinal NAD(P) quinone oxidoreductase 1 expression and decreased oxidative stress-related ferritin, ionized calcium-binding adapter molecule 1-immunoreactive microglia, S100β-immunoreactive astrocyte, and Olig2-immunoreactive oligodendrocytes compared with the control vehicle. Interestingly, we found that the spinal LPA1 receptor level was decreased, whereas gintonin treatment restored decreased LPA1 receptor expression levels in the G93A-SOD1 transgenic mouse, thereby attenuating neurological symptoms and histological deficits. Conclusion: Gintonin-mediated symptomatic improvements of ALS might be associated with the attenuations of neuronal loss and oxidative stress via the spinal LPA1 receptor regulations. The present results suggest that the spinal LPA1 receptor is engaged in ALS, and gintonin may be useful for relieving ALS symptoms.

• The Korea Journal of Herbology
• /
• v.29 no.3
• /
• pp.27-33
• /
• 2014

Objectives : The aim of this study was to investigate the protective effect of extract of Thuja orientalis leaves (TOFE) against 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced neurotoxicity by inhibition of inflammation in in vitro and in vivo models of Parkinson's disease (PD). Methods : We evaluated the effect of TOFE against lipopolysaccharide (LPS)/1-methyl-4-phenylpyridinium ($MPP^+$) toxicity using nitric oxide (NO) assay, inducible NO synthase and cyclooxygenase 2 western blot, tyrosine hydroxylase and microglia activation immunohistochemistry (IHC) in BV2 cell, primary rat mesencephalic neurons, or C57BL/6 mice. We also evaluated the effect of TOFE in mice PD model induced by MPTP. C57BL/6 mice were treated with TOFE 50 mg/kg for 5 days and were injected intraperitoneally with four administrations of MPTP on the last day. We conducted behavioral tests and IHC analysis to see how TOFE affect MPTP-induced neuronal loss of dopaminergic neurons in substantia nigra pars compacta (SNpc) and striatum (ST) of mice. To assess the anti-inflammation effects, we carried out glial fibrillary acidic protein and macrophage-1 antigen integrin alpha M in IHC in SNpc and ST of mice. Results : In an in vitro system, TOFE decreasesd NO generations in BV2 cells. TOFE protected dopaminergic cells against LPS or $MPP^+$-induced toxicity in primary mesencephalic dopaminergic neurons. In vivo system, TOFE at 50 mg/kg treated group showed improved motor deteriorations than the MPTP only treated group and TOFE significantly protected striatal dopaminergic damage from MPTP-induced neurotoxicity in mice. Moreover, TOFE inhibited activation of astrocyte and microglia in SNpc and ST of the mice. Conclusions : We concluded that TOFE showed anti-parkinsonian effect by protection of dopaminergic neurons against MPTP toxicity through anti-inflammatory actions.

• The Journal of the Acoustical Society of Korea
• /
• v.42 no.5
• /
• pp.412-421
• /
• 2023

Ultrasound brain stimulation is spot-lighted by its capability of inducing brain cell activation in a localized deep brain region and ultimately treating impaired brain function while the efficiency and directivity of neural modulation are highly dependent on types of stimulus waveforms. Therefore, to optimize the types of stimulation parameters, we propose a cell-cultivable ultrasonic transducer having a series stack of a spin-coated polymer piezoelectric element (Poly-vinylidene fluoride-trifluorethylene, PVDF-TrFE) and a parylene insulating layer enhancing output acoustic pressure on a glass-coverslip which is commonly used in culturing cells. Due to the uniformity and high accuracy of stimulus waveform, tens of neuronal cell responses located on the transducer surface can be recorded simultaneously with fluorescence microscopy. By averaging the cell response traces from tens of cells, small changes to the low intensity ultrasound stimulations can be identified. In addition, the reduction of stimulus distortions made by standing wave generated from reflections between the transducers and other strong reflectors can be achieved by placing acoustic absorbers. Through the proposed ultrasound transducer, we could successfully observe the calcium responses induced by low-intensity ultrasound stimulation of 6 MHz, 0.2 MPa in astrocytes cultured on the transducer surface.

• Development and Reproduction
• /
• v.12 no.1
• /
• pp.31-39
• /
• 2008

Neural tissue has limited intrinsic capacity of repair after injury, and the identification of alternate sources of neural stem cells has broad clinical potential. We isolated mesechymal-like stem cells from human adipose tissues (AT-MSCs), and studied on transdifferentiation-promoting conditions in neural cells. Dopaminergic and cholinergic neuron induction of AT-MSCs was also studied. Neural differentiation was induced by adding bFGF, EGF, dimethyl sulphoxide (DMSO) and butylated hydroxyanisole(BHA) in N2 Medium and N2 supplement. The immunoreactive cells for $\beta$-tubulin III, a neuron-specific marker, GFAP, an astrocyte marker, or Gal-C, an oligodendrocyte marker, were found. AT-MSCs treated with bFGF, SHH and FGF8 were differentiatied into dopaminergic neurons that were immunopositive for TH antibody. Differentiation of MSCs to cholinergic neurons was induced by combined treatment with basic fibroblast growth factor (bFGF), retinoic acid (RA) and sonic hedgehog (Shh). AT-MSCs treated with DMSO and BHA rapidly assumed the morphology of multipolar neurons. Both immunocytochemistry and RT-PCR analysis indicated that the expression of a number of neural markers including neuro D1, $\beta$-tubulin III, GFAP and nestinwas markedly elevated during this acute differentiation. While the stem cell markers such as SCF, C-kit, and Stat-3 were not expressed after preinduction medium culture, we confirmed the differentiation of dopaminergic and cholinergic neurons by TH/$\beta$-tubulin III or ChAT/ $\beta$-tubulin III positive cells. Conclusively, AT-MSCs can be differentiated into dopaminergic and cholinergic neuronsand these findings suggest that AT-MSCs are alternative cell source of treatment for neurodegenerative diseases.

• Journal of Life Science
• /
• v.16 no.5
• /
• pp.840-849
• /
• 2006

Severe brain injuries induced by toxin pose one of the most important problems on our health care because of their high morbidity and mortality, are implicated to leucocyte infiltration more premature or immature brain than mature brain. Chemokines are the induction meditators for infiltration of inflammatory cells to the inflammation sites. In order to study the mechanism of leucocyte infiltration, the expression of several chemokines, MCP-1, $MIP-1{\alpha}$ and MIP-2 was studied in lipopolysaccharide(LPS)-stimulated neonatal and adult brain. One week old Sprague-Dawley rats or adult male rats weighing 300-350 g were used for the experiment. After anesthetization, $1\;{\mu}l$ LPS (0.5 mg/ml) subsequently was injected in the right caudate nucleus of the brain with stereotaxic frame. Animals were sacrificed at 6 hours, 24 hours, and 72 hours after injection. The present study was carried out using RT-PCR for the mRNA and immunohistochemistry for the expression of the proteins. In the neonatal rat brain, prominent interstitial edema with significant accumulation of leukocytes was detected at 24 and 72 hours after LPS injection. A semiquantitative analysis of RT-PCR revealed that the MCP-1, $MIP-1{\alpha}$, and MIP-2 mRNA expression peaked at 24 hours in neonatal and adult rat brain. Neonatal rats showed about 2.6, 1.4, and 1.2 times more expression of the MCP-1, $MIP-1{\alpha}$, and MIP-2 than that of the adult rats in the brain tissue. Immunohistochemical analysis also showed that MCP-1 immunoreactivity was paralleled with the RT-PCR results. MCP-1 protein was significantly detected at 24 and 72 hours in the brain parenchyma. $MIP-1{\alpha}$protein was highly expressed at 24 hours. The results of leukocyte infiltration in H&E stain was parallelled with that of the immunohistochemistry. Chemokine proteins were markedly detected at 24 hours after injection of LPS and neutrophil influx into intraparenchymal was prominent at 24 hours. These results suggest that the leukocyte infiltration in the intracranial infection may be controlled by mechanisms influenced by chemokine producing cells in the central nervous system such as microglia, astrocyte and endothelial cell.