• 한국축산식품학회지
• /
• 제40권5호
• /
• pp.852-859
• /
• 2020

The muscle stem cells of domestic animals are of interest to researchers in the food and biotechnology industries for the production of cultured meat. For producing cultured meat, it is crucial for muscle stem cells to be efficiently isolated and stably maintained in vitro on a large scale. In the present study, we aimed to optimize the method for the enrichment of pig muscle stem cells using a magnetic-activated cell sorting (MACS) system. Pig muscle stem cells were collected from the biceps femoris muscles of 14 d-old pigs of three breeds [Landrace×Yorkshire×Duroc (LYD), Berkshire, and Korean native pigs] and cultured in skeletal muscle growth medium-2 (SkGM-2) supplemented with epidermal growth factor (EGF), dexamethasone, and a p38 inhibitor (SB203580). Approximately 30% of total cultured cells were nonmyogenic cells in the absence of purification in our system, as determined by immunostaining for cluster of differentiation 56 (CD56) and CD29, which are known markers of muscle stem cells. Interestingly, following MACS isolation using the CD29 antibody, the proportion of CD56⁺/CD29⁺ muscle stem cells was significantly increased (91.5±2.40%), and the proportion of CD56 single-positive nonmyogenic cells was dramatically decreased. Furthermore, we verified that this method worked well for purifying muscle stem cells in the three pig breeds. Accordingly, we found that CD29 is a valuable candidate among the various marker genes for the isolation of pig muscle stem cells and developed a simple sorting method based on a single antibody to this protein.

• Journal of Applied Biological Chemistry
• /
• 제59권4호
• /
• pp.285-288
• /
• 2016

단백질 분해효소는 단백질 내 아미노산의 펩티드 결합을 가수분해하며, 산업분야에서 가장 많이 사용되는 효소이다. 이전의 논문에서 부패한 나무로부터 Pseudoxanthomonas sp. WD12와 WD32를 분리하였다. 분비된 단백질 분해효소의 특성을 조사한 결과 WD12는 pH 9.0, $50^{\circ}C$, WD32의 경우 pH 8.0, $45^{\circ}C$에서 최적 활성을 나타내었다. 최적조건하에서 최고의 활성은 WD12는 768 unit/mL로 WD12의 활성이 WD32보다 2.6배 높았다. 금속 이온에 대한 분비효소 영향을 조사한 결과 두 균주 모두 KCl, NaCl, $CaCl_2$, $MnSO_4$를 첨가했을 때 활성이 증가하였으며, $AgNO_3$, $CuSO_4$, $FeCl_3$, $AlCl_3$를 첨가했을 때는 감소하였고, 최고의 활성은 $MnSO_4$의 첨가했을 때 두 균주 모두 활성 증가를 보인 반면 $FeCl_3$, $CuSO_4$는 활성저해를 보였다. 신종으로 생각되는 WD12 균주의 효소는 알칼리성 조건의 산업 환경에서 이용가능 하다고 생각된다.

• The Korean Journal of Physiology and Pharmacology
• /
• 제17권3호
• /
• pp.203-208
• /
• 2013

As the abnormal proliferation of vascular smooth muscle cells (VSMCs) plays a critical role in the development of atherosclerosis and vascular restenosis, a candidate drug with antiproliferative properties is needed. We investigated the antiproliferative action and underlying mechanism of a newly synthesized naphthoquinone derivative, 5,8-dimethoxy-2-nonylamino-naphthalene-1,4-dione (2-nonylamino-DMNQ), using VSMCs treated with platelet-derived growth factor (PDGF). 2-Nonylamino-DMNQ inhibited proliferation and cell number of VSMCs induced by PDGF, but not epidermal growth factor (EGF), in a concentration-dependent manner without any cytotoxicity. This derivative suppressed PDGF-induced $[^3H]$-thymidine incorporation, cell cycle progression from $G_0/G_1$ to S phase, and the phosphorylation of phosphor-retinoblastoma protein (pRb) as well as the expression of cyclin E/D, cyclin-dependent kinase (CDK) 2/4, and proliferating cell nuclear antigen (PCNA). Importantly, 2-nonylamino-DMNQ inhibited the phosphorylation of PDGF receptor${\beta}$(PDGF-$R{\beta}$) enhanced by PDGF at $Tyr^{579}$, $Tyr^{716}$, $Tyr^{751}$, and $Tyr^{1021}$ residues. Subsequently, 2-nonylamino-DMNQ inhibited PDGF-induced phosphorylation of STAT3, ERK1/2, Akt, and $PLC{\gamma}1$. Therefore, our results indicate that 2-nonylamino-DMNQ inhibits PDGF-induced VSMC proliferation by blocking PDGF-$R{\beta}$ autophosphorylation, and subsequently PDGF-$R{\beta}$-mediated downstream signaling pathways.

• 한국식품영양과학회지
• /
• 제40권5호
• /
• pp.635-641
• /
• 2011

본 연구는 TNF-${\alpha}$로 자극된 HUVEC에서 녹차씨껍질 EtOAC 추출물이 초기 동맥경화 과정에 중요한 역할을 하는 염증매개인자와 세포부착물질에 미치는 영향을 분석하였다. 녹차씨껍질 EtOAC 추출물의 NO 생성능은 TNF-${\alpha}$만을 처리한 control군에 비해 증가시키는 것을 알 수 있었다. $100{\mu}g$/mL 농도에서는 녹차씨껍질 EtOAC 추출물은 세포독성을 보이지 않았으며 염증매개인자인 TNF-${\alpha}$ 수준 및 세포부착물질인 VCAM-1과 MCP-1의 생성을 억제하였다. 뿐만아니라, 녹차씨껍질 EtOAC 추출물은 총 항산화능은 증가되는 경향을 보였다. 이상의 결과에서, 녹차씨껍질 EtOAC 추출물은 HUVEC에서 TNF-${\alpha}$로 인한 총 항산화능의 수준을 향상시켜 염증생성인자인 TNF-${\alpha}$ 수준 및 세포부착물질인 VCAM-1과 MCP-1의 생성을 억제하여 동맥경화 초기반응을 억제하는데 기여할 것으로 사료된다.

• Journal of Ginseng Research
• /
• 제42권4호
• /
• pp.401-411
• /
• 2018

Longevity in medicine can be defined as a long life without mental or physical deficits. This can be prevented by Alzheimer's disease (AD). Current conventional AD treatments only alleviate the symptoms without reversing AD progression. Recent studies demonstrated that Panax ginseng extract improves AD symptoms in patients with AD, and the two main components of ginseng might contribute to AD amelioration. Ginsenosides show various AD-related neuroprotective effects. Gintonin is a newly identified ginseng constituent that contains lysophosphatidic acids and attenuates AD-related brain neuropathies. Ginsenosides decrease amyloid ${\beta}$-protein ($A{\beta}$) formation by inhibiting ${\beta}$- and ${\gamma}$-secretase activity or by activating the nonamyloidogenic pathway, inhibit acetylcholinesterase activity and $A{\beta}$-induced neurotoxicity, and decrease $A{\beta}$-induced production of reactive oxygen species and neuro-inflammatory reactions. Oral administration of ginsenosides increases the expression levels of enzymes involved in acetylcholine synthesis in the brain and alleviates $A{\beta}$-induced cholinergic deficits in AD models. Similarly, gintonin inhibits $A{\beta}$-induced neurotoxicity and activates the nonamyloidogenic pathway to reduce $A{\beta}$ formation and to increase acetylcholine and choline acetyltransferase expression in the brain through lysophosphatidic acid receptors. Oral administration of gintonin attenuates brain amyloid plaque deposits, boosting hippocampal cholinergic systems and neurogenesis, thereby ameliorating learning and memory impairments. It also improves cognitive functions in patients with AD. Ginsenosides and gintonin attenuate AD-related neuropathology through multiple routes. This review focuses research demonstrating that ginseng constituents could be a candidate as an adjuvant for AD treatment. However, clinical investigations including efficacy and tolerability analyses may be necessary for the clinical acceptance of ginseng components in combination with conventional AD drugs.

• 대한본초학회지
• /
• 제32권1호
• /
• pp.25-31
• /
• 2017

Objectives : Mori Radicis Cortex (MRC), the root epidermis of Morus alba L., has been traditionally used to treat lung-related diseases in Korean Medicine. The common of MRC is Mulberry bark Morus bark, and it's pharmaceutical properties and taste are known as sweet and cold, and it promotes urination and reduce edema by reducing heat from the lungs and soothe asthma. In the present study, anti-apoptotic mechanism of MRC in middle cerebral artery occlusion (MCAO) model in mice. Methods : Two-hundred grams of MRC was extracted with methanol at room temperature for 5 days, and this was repeated one time. After filtration, the methanol was removed using vacuum evaporator, then stored at $-20^{\circ}C$ until use. C57BL/6 male mice were housed in an environment with controlled humidity, temperature, and light cycle. In order to determine beneficial effects of MRC on ischemia induced brain damage, infarct volume, neurological deficit scores, activities of several apoptosis-related proteins such as caspase-8, -9, Bcl-xL in MCAO-induced brains of mice were analyzed. Mice in MRC-treated groups were orally administered 30, 100, or 300 mg/kg of body weight for three consecutive days before commencing the MCAO procedure. Results : Pre-treatment of MRC significantly reduced infarct volume in MCAO subjected mice applied with 300 mg/kg of MRC methanol extract, and MRC effectively inhibited Bcl-xL reduction and caspase-9 activation caused by MCAO-induced brain damage. Conclusions : MRC showed neuro-protective effects by regulating apoptosis-related protein signals, and it can be a potential candidate for the therapy of ischemia-induced brain damage.

• Archives of Pharmacal Research
• /
• 제25권2호
• /
• pp.170-177
• /
• 2002

The herb, Chrysanthemum zawadskii var, latilobum commonly known as Gu-Jul-Cho in Korea, used in traditional medicine to treat pneumonia, bronchitis, cough, common cold, pharyngitis, bladder-related disorders, gastroenteric disorders, and hypertension. Linarin is the main active compound and the biological mechanisms of its activity are unclear. It is believed that effects of this herb may be exerted through the pluripotent effectors of linarin due to its ability to treat a variety of afflictions. In this study, the effects of linarin on the mouse macrophages cell line, RAW 264.7, were investigated. It was found that linarin could activate macrophages by producing cytokines. Monocytes and tissue macrophages produce at least two groups of protein mediators of inflammation, interleukin 1 (IL-1 ) and the tumor necrosis factor (TNF). Recent studies have shown that TNF and IL-1 modulate the inflammatory function of endothelial cells, leukocytes, and fibroblasts. $TNF-{\alpha}$ production by macrophages treated with linarin occured in a dose dependent manner However, IL-1 production was largely unaffected by this natural product. This study demonstrated the ability of linarin to activate macrophages both directly and indirectly. Linarin also affect both cytosine production and nitric oxide inhibition, in addition to the expression of some surface molecules. Nitric oxide (NO), derived from L-argin-ine, is produced by two forms(constitutive and inducible) of nitric oxide synthase (NOS). The NO produced in large amounts by inducible NOS is known to be responsible for the vasodilation and hypotension observed in septic shock. Linarin was found to inhibit NO production in the LPS-activated RAW 264.7 cells. Linarin may be a useful candidate as a new drug for treating endotoxemia and the inflammation accompanied by NO overproduction. The linarin-treated total Iymphocytes exhibited cytotoxicity in a dose dependent manner between $20{\;}{\mu}g/ml{\;}and{\;}40{\;}{\mu}g/ml$. These results suggest that linarin may function through macrophage activation.

• 생명과학회지
• /
• 제22권10호
• /
• pp.1399-1406
• /
• 2012

본 연구에서는 톳 분획물의 항비만 효과 및 그에 따른 생화학적 기전의 해석을 위하여 톳 분획물이 비만유도인자에 의하여 인위적으로 유발된 adipogenesis 과정에 있어서 어떠한 영향을 미치는 지를 조사하였고, 이때 $PPAR{\gamma}$, C/$EBP{\alpha}$ 및 C/$EBP{\beta}$ 등과 같은 adipogenic transcription factor들의 발현에 어떠한 변화가 유발되었는지를 조사하였다. 각각의 톳 분획물들이 성숙한 지방세포에서 나타나는 lipid droplet 및 TG 생성에 어떠한 영향을 미치는 지를 확인한 결과, 모든 분획물에서 lipid droplet 및 TG 생성억제가 나타났지만 특히 WFHF 처리군에서 이러한 현상이 가장 강하게 나타났다. 또한 lipid droplet 및 TG 생성에 중요한 역할을 하는 것으로 알려진 adipogenic transcription factor들의발현에각각의분획물들이 어떠한영향을미치는지를확인한결과,WFHF 처리군에서 $PPAR{\gamma}$, C/$EBP{\alpha}$ 및 C/$EBP{\beta}$의 발현이 현저하게 감소하였음을 확인하였다. 이상의 결과를 종합해 보면 다섯 종류의 톳 분획물 모두 비만억제 효과가 있는 것으로 나타났고 특히 WFHF의 비만억제 효과가 강하게 나타났음을 알 수 있었다. 본 연구 결과는 톳의 비만억제 가능성을 제시하는 것으로서 항비만 기전에 대한 생화학적 해석 및 이를 활용한 향후 지속적인 연구를 위한 자료로서 그 가치가 매우 높을 것으로 생각된다.

• 한국식품과학회지
• /
• 제45권1호
• /
• pp.90-96
• /
• 2013

소리쟁이 추출물 및 분획물의 3T3-L1 전지방세포의 분화에 미치는 영향을 확인한 결과, 소리쟁이의 hexane, ethyl acetate 분획물에 의해 지방세포의 lipid droplet의 형성이 유의적으로 억제되었으며, 소리쟁이 에탄올 추출물과 모든 분획물 10 ${\mu}g/mL$의 농도에서 중성지질(triglyceride)의 함량이 유의적으로 감소되었다. 또한 지방세포의 분화에 관여하는 전사인자인 $PPAR{\gamma}$, $C/EBP{\alpha}$의 단백질 발현은 10 ${\mu}g/mL$의 ethyl acetate, butanol 분획물에 의해 현저하게 감소되었다. 따라서 소리쟁이 추출물과 분획물 중 가장 활성이 우수한 ethyl acetate 분획물을 이용하여 지방분화에 관여하는 전사 인자인 $PPAR{\gamma}$, $C/EBP{\alpha}$, 그리고 SREBP1c 및 지질의 합성, 수송, 저장에 관여하는 ACS, FAS, FATP1, FABP4, Perilipin의 발현에 미치는 영향을 관찰한 결과, ethyl acetate 분획물은 유의적으로 모든 유전자의 발현을 억제시켰다. 따라서 소리쟁이의 ethyl acetate 분획물은 지방세포의 분화에 관여하는 전사인자 및 유전자들의 발현을 감소시킴으로써 항비만 효과가 있는 천연물 소재로 이용 가능할 것으로 생각된다.

• 생명과학회지
• /
• 제29권11호
• /
• pp.1200-1207
• /
• 2019

Loganin은 Corni fructus의 주요 iridoid glycoside이며 항염증, 항당뇨 그리고 뇌신경보호 효과 등이 보고되었다. 본 연구에서는 ${\alpha}-MSH$와 IBMX처리된 B16F10세포에서 loganin의 melanogenesis억제효과의 신호전달 경로를 조사하였다. Loganin의 미백 활성을 확인하기 위해 B16F10세포에서 $1{\mu}m$에서 $20{\mu}m$사이의 농도를 처리하여 세포독성 실험을 수행한 결과 최대 $20{\mu}m$농도에서 독성을 나타내지 않았다. 또한 loganin은 ${\alpha}-MSH$와 IBMX처리된 B16F10세포에서 농도-의존적으로 멜라닌 생성을 감소시키는 것을 확인하였다. 또한 loganin의 멜라닌 생성을 억제하는 신호전달 경로를 Western blotting을 실시하여 조사하였다. Western blot결과에 따르면 loganin은 ${\alpha}-MSH$와 IBMX 처리된 B16F10세포에서 증가된 CREB인산화(Ser133)와 MITF 발현 및 tyrosinase의 유전자 발현을 감소시켰고 ERK의 인산화를 증가시켜 melanin 생성을 억제하였다. 결론적으로 loganin은 ${\alpha}-MSH$와 IBMX에 의해 유도된 과도한 멜라닌 합성을 CREB인산화와 MITF 및 tyrosinase의 유전자 발현을 억제하고 ERK의 활성화를 통해 멜라닌 합성을 감소됨을 확인하였다. 따라서 loganin은 과색소 침착과 관련된 피부질환의 보호제로서 활용될 가능성을 가지는 것으로 사료된다.