• 한국미생물·생명공학회지
• /
• 제25권1호
• /
• pp.102-105
• /
• 1997

In cultivating human neuroblastoma cells maximum number of neurites per cell and length of the neurite were estimated as 5.5 and 2.2 (nm), respectively It was found that there was correlation between growth and differentiation of nerve cells. Maximum specific BDNF production rate was also calculated as 2.5$\times $10$^{-5}$(ng/cell/day) at 7$\times $ 10$^{5}$ (viable cells/ml) of maximum cell density, corresponding to 100 (ng/mL) of BDNF. The secretion of BDNF was occurred most in the later peroids of the cultivation, yielding 75 (ng/mL) of BDNF. The production of rate of BDNF was elongated in adding 1 ($\mu $g/mL) of BDNF as well as 40% increase of the length of the BDNF. It proves that BDNF can be used as one of biopharmaceuticals to treat age-related diseases such as Alzheimer's disease and Prakinson's disease. It can also provide the information of scaling-up mammalian cell cuture system to economically produce BDNF.

• KSBB Journal
• /
• 제18권3호
• /
• pp.207-210
• /
• 2003

Locus coeruleus (LC)에는 전체 노르아드레날린성 뉴런의 절반 가량이 모여 있는데, 여기서 노르아드레날린성 뉴런이 뇌의 거의 모든 부위로 신경자극을 보내게 된다. LC는 알츠하이머병, 파킨슨병, 헌팅턴병 같은 여러 가지 신경퇴행성 질환에서 공통적으로 타격을 받는 주요 부위이다. 뇌 유래 신경영양인자, BDNF가 LC 노르아드레날린성 뉴런을 포함한 중추신경계 뉴런들의 분화와 신경세포 생존에 중요한 조절자로 작용한다. 본 연구에서는 LC 노르아드레날린성 신경세포에서 여러 가지 작은 분자들과 성장인자들이 BDNF 생산을 촉진할 수 있는지를 조사하였다. 실험에 사용한 분자들로는 neuropeptides, cytokines, 성장인자, 신경전달물질들과 세포내 신호전달물질들이 포함되었다. 여러 가지 작은 분자들과 성장인자들 중에서 FGF8b, BMP-4, forskolin 그리고 dibutyrl cGMP가 LC 노르아드레날린성 뉴런에서 BDNF 분비를 뚜렷하게 증대시킨 것으로 판명되었다. 특히, BMP-4는 BDNF 생산을 2.5배 이상 증가시켰다. LC 노르아드레날린성 뉴런에서 BDNF를 증가시킨 물질들은 여러 가지 신경퇴행성 질환에서 신경세포가 손실되는 것을 막거나 지연시킬 수 있을 것이므로, 치료제나 증상완화제로서의 가능성이 높다.

• Biomolecules & Therapeutics
• /
• 제20권1호
• /
• pp.27-35
• /
• 2012

Oroxylin A is a flavone isolated from a medicinal herb reported to be effective in reducing the inflammatory and oxidative stresses. It also modulates the production of brain derived neurotrophic factor (BDNF) in cortical neurons by the transactivation of cAMP response element-binding protein (CREB). As a neurotrophin, BDNF plays roles in neuronal development, differentiation, synaptogenesis, and neural protection from the harmful stimuli. Adenosine $A2_A$ receptor colocalized with BDNF in brain and the functional interaction between $A2_A$ receptor stimulation and BDNF action has been suggested. In this study, we investigated the possibility that oroxylin A modulates BDNF production in cortical neuron through the regulation of $A2_A$ receptor system. As expected, CGS21680 ($A2_A$ receptor agonist) induced BDNF expression and release, however, an antagonist, ZM241385, prevented oroxylin A-induced increase in BDNF production. Oroxylin A activated the PI3K-Akt-GSK-$3{\beta}$ signaling pathway, which is inhibited by ZM241385 and the blockade of the signaling pathway abolished the increase in BDNF production. The physiological roles of oroxylin A-induced BDNF production were demonstrated by the increased neurite extension as well as synapse formation from neurons. Overall, oroxylin A might regulate BDNF production in cortical neuron through $A2_A$ receptor stimulation, which promotes cellular survival, synapse formation and neurite extension.

• Journal of Microbiology and Biotechnology
• /
• 제9권3호
• /
• pp.323-327
• /
• 1999

It was shown that brain-derived neurotrophic factors (BDNFs) secreted from human neuroblastoma cells can significantly improve the growth of the neurites of PC12 nerve cells. The addition of purified BDNFs elongated the neurites of PC 12 nerve cells two to three times more than the case where the addition was not made. The perfusion rate strongly affected the change of the size of human neuroblastoma cells because the cell size decreased as the perfusion rate increased. This could also influence the productivity of BDNF from the cells. It is also important to note that the BDNF production was decreased when the cell size was reduced. BDNF production rate also decreased at a fast perfusion rate in a smaller cell size. At the relatively fast perfusion rate of 18 ml/h, the ratio of apoptotic to necrotic cells dramatically decreased, which possibly caused the decrease of BDNF production. It has been proven that the secretion of BDNF from human neuroblastoma cells was a partially growth-related process by yielding 6.2$\times l0^{-8}/g$ of BDNF/cell/h of growth related parameter and $0.48{\times}l0^{-9}/g$ of BDNF/cell/h of nongrowth-related parameter in a growth kinetic model. In addition, it was also found that the perfusion rate played a very important role in controlling the cell death mechanism.

• Biotechnology and Bioprocess Engineering:BBE
• /
• 제3권2호
• /
• pp.51-54
• /
• 1998

It has been proved that co-cultivation of human neroblastoma cells and human fibroblast cells can enhance nerve cell growth and the production of BDNF in perfusion cultivation. In batch co-cultivation, maximum cell density was increased up to 1.76${\times}$106 viable cells/mL from 9${\times}$105 viable cells/mL of only neuroblastoma cell culture. The growth of neuroblastoma cells was greatly improved by culturing both nerve and fibroblast cells in a perfusion process, maintaining 1.5${\times}$106 viable cells/mL, which was much higher than that form fed-batch cultivation. The nerve cell growth was greatly enhance in both fed-batch and perfusion cultivations while the growth of fibroblast cells was not. It strongly implies that the factors secreted from human fibrobast cells and/or the environments of co-culture system can enhance both cell growth and BDNF secretion. Specific BDNF production rate was not enhanced in co-cultures; however, the production period was increased as the cell growth was lengthened in the co-culture case. Competitive growth between nerve cells and fibroblast cells was not observed in all cases, showing no changes of fibroblast cell growth and only enhancement of the neuroblastoma cell growth and overall BDNF production. It was also found that the perfusion cultivation was the most appropriate process for cultivating two cell lines simultaneously in a bioreactor.

• Journal of Microbiology and Biotechnology
• /
• 제29권9호
• /
• pp.1369-1374
• /
• 2019

We isolated Lactobacillus mucosae NK41 and Bifidobacterium longum NK46 from human feces, which induced BDNF expression in corticosterone-stimulated SH-SY5Y cells, and examined their anti-depressive effects in mice. NK41, NK46, and their (1:1) mixture significantly mitigated immobilization stress (IS)-induced anxiety-like/depressive behaviors, hippocampal $NF-{\kappa}B$ activation, BDNF expression, $Iba1^+$ cell population, and blood corticosterone, $TNF-{\alpha}$, IL-6, and lipopolysaccharide levels. Furthermore, they inhibited colitis marker $NF-{\kappa}B$ activation, and $TNF-{\alpha}$ expression in mice with IS-induced anxiety/depression. They additionally suppressed gut Proteobacteria and Bacteroidetes populations and bacterial lipopolysaccharide production. These findings suggest that NK41 and NK46 may alleviate anxiety/depression and colitis by suppressing gut dysbiosis.

• 대한방사선기술학회지:방사선기술과학
• /
• 제36권3호
• /
• pp.209-217
• /
• 2013

이 연구는 지방조직에서 분비되어 비만의 진행 조절에 관여하는 adiponectin과 leptin의 복부지방두께와의 관련성을 보고자 하였다. 연구대상자는 남성 근로자 138명으로 초음파를 이용하여 피하 및 내장지방두께를 측정하고 ELISA kit을 이용하여 adiponectin, BDNF 및 leptin의 농도를 측정하였다. 그 결과, 대상자의 평균피하지방과 내장지방두께는 $1.58{\pm}0.51$과 $4.52{\pm}1.44cm$이었다. Adiponectin, BDNF 및 leptine의 평균농도는 각각 $3.14{\pm}3.52$ ng/ml, $24.11{\pm}8.52$ pg/ml와 $4.27{\pm}2.38$ ng/ml이었다. 내장지방두께는 총 콜레스테롤, LDL-콜레스테롤, 중성지방 및 인슐린 농도와 양의 상관관계를 보였으나, HDL-콜레스테롤과는 음의 상관관계를 보였다. Adiponectin 농도는 HDL-콜레스테롤의 농도와 양의 상관관계를 보였고 총 콜레스테롤, LDL-콜레스테롤, 중성지방, 피하 및 내장지방두께와는 음의 상관관계를 보였다. 그러나 leptin의 농도는 adiponectin과 상반되는 결과를 보였다. Adiponectin의 농도는 비만하지 않은 대상자(BMI<25 $kg/m^2$)에서 유의하게 높았으나 leptin의 농도는 비만한 대상자((BMI>25 $kg/m^2$)에서 높았다. 비만과 adiponectin, BDNF 및 leptin의 관련성을 보기 위하여, 연령, 흡연 및 음주습관 등을 보정한 다음, 다중 로지스틱 회귀분석을 실시한 결과, 비만은 adiponectin(교차비=0.784, p=0.006)과 leptin(교차비=1.493, p=0.001)이 관련이 있는 것으로 나타났다. 이상의 결과는 adiponectin과 leptin의 농도가 복부지방에 영향을 주고, 이들 신경영양 물질들의 생성과 분비 기능저하는 궁극적으로 비만과 심혈관계 질환의 유발에 관여함을 보여준 결과라 판단된다.

• Fisheries and Aquatic Sciences
• /
• 제26권1호
• /
• pp.24-34
• /
• 2023

Reactive oxygen species (ROS) at high concentrations induce oxidative stress, an imbalanced redox state that is a prevalent cause of neurodegenerative disorders. This study aimed to investigate the protective effect of Capsosiphon fulvescens (CF) extract on oxidative stress-induced impairment of cognitive function in models of neurodegenerative diseases. CF was extracted with subcritical water and several solvents and H₂O₂ (0.25 mM) or aluminum chloride (AlCl₃; 25 µM) as an inducer of ROS was treated in PC12 neuronal cells and zebrafish larvae. All statistical analyses were performed using one-way analysis of variance and Dunnett's test using GraphPad Prism. H₂O₂ and AlCl₃ were found to significantly induce ROS production in PC12 neuronal cells and zebrafish larvae. In addition, they strongly affected intracellular Ca²⁺ levels, antioxidant enzyme activity, brain-derived neurotrophic factor (BDNF) and tropomyosin receptor kinase B (TrkB) signaling, acetylcholinesterase (AChE) activity, and hallmarks of Alzheimer's disease. However, treatment of H₂O₂-induced PC12 cells or AlCl₃-induced zebrafish larvae with CF subcritical water extract at 90℃ and CF water extract effectively regulated excessive ROS production, intracellular Ca²⁺ levels, and mRNA expression of superoxide dismutase, glutathione peroxide, glycogen synthase kinase-3 beta, β-amyloid, tau, AChE, BDNF, and TrkB. Our study suggested that CF extracts can be a potential source of nutraceuticals that can improve the impairment of cognitive function and synaptic plasticity by regulating ROS generation in neurodegenerative diseases.

• 한국동물번식학회:학술대회논문집
• /
• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
• /
• pp.79-79
• /
• 2003

Embryonic stem cells are capable of differentiating into a variety of cell lineages. However, the ultimate results of differentiation in vitro greatly depend on the duration of treatment and kinds of differentiating inducers added. In order to investigate the efficiencies of various differentiation inducers and the methods of treatment, we examined differentiation patterns of human embryonic stem cell (hESC, MB03) according to several different protocols. Exp. I) Upon differentiation using retinoic acid and ascorbic acid (RA/AA), embryoid bodies (EB, for 4days) derived from hESC was exposed to Rh (10$^{-6}$ M) and AA (50 mM) for 4 days, and were allowed to differentiate in N2 medium for 7, 14, 21, or 28 days. Exp. II) When bFGF was used, neuronal precursor cells were selected for 8 days in N2 medium after EB formation. After selection, cells were expanded at the presence of bFGF (20 ng/ml) for another 6 days followed by a final differentiation in N2 medium for 7, 14, 21 or 28 days. Exp. III) In addition, to examine the effects of neurotrophic factors in the production of mature neurons, groups of cells were exposed to either BDNF (5 ng/ml) or TGF-$\alpha$(10 ng/ml) during the 28 days of final differentiation. Differentiation patterns of RA/AA or bFGF treated groups were very similar; approximately 82% and 83% of the cells, respectively, were positive for anti-NF200 antibody, while it was about 10% and 11%, respectively, for anti-NF160 antibody in 28 days in N2 medium. Alsor, cells expressing TH were as low as 5%, while the cells doubled when matured at the presence of either BDNF or TGF-$\alpha$. Cells immunoreactive to anti-GAD antibody were approximately 20%. These results suggest that a maturation step rather than differentiation induction step, which is formation of EB, effects more decisively to the ultimate differentiation pattern.

• Biomolecules & Therapeutics
• /
• 제24권5호
• /
• pp.469-474
• /
• 2016

Liriopogons (Liriope and Opiopogon) species are used as a main medicinal ingredient in several Asian countries. The Liriopes Radix (tuber, root of Liriope platyphylla) has to be a promising candidate due to their source of phytochemicals. Steroidal saponins and their glycosides, phenolic compounds, secondary metabolites are considered of active constituents in Liriopes Radix. Spicatoside A, a steroidal saponin, could be more efficacious drug candidate in future. In this review, we summarized the available knowledge on phytochemical and pharmacological activities for spicatoside A. It significantly suppressed the level of NF-${\kappa}B$, NO, iNOS, Cox-2, IL-$1{\beta}$, IL-6 and MAPKs in LPS-stimulated inflammation. The production of MUC5AC mucin was increased. MMP-13 expression was down-regulated in IL-$1{\beta}$-treated cells and reduced glycosaminoglycan release from IL-$1{\alpha}$-treated cells. The neurite outgrowth activity, PI3K, Akt, ERK1/2, TrkA and CREB phosphorylation and neurotropic factors such as NGF and BDNF were upregulated with increased latency time. It also showed cell growth inhibitory activity on various carcinoma cells. From this, spicatoside A exerts anti-inflammation, anti-asthma, anti-osteoclastogenesis, neurite outgrowth, memory consolidation and anticancer activities. Further studies are needed on spicatoside A in order to understand mechanisms of action to treat various human diseases.