• 생명과학회지
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• 제29권2호
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• pp.191-197
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• 2019

파킨슨병은 운동완서, 근육경직, 진전 및 비정상적인 자세 등을 임상적 특징으로 하는 주로 운동 신경계에 영향을 주는 진행성 신경 퇴행성 질환이다. 파킨슨병은 산화 스트레스와 세포 내 신호 전달 경로의 조절 장애에 의한 뇌 흑색치밀부에서의 도파민성 신경세포의 사멸을 특징으로 한다. Quercetin의 주요 대사산물인 Quercetin-3-O-glucuronide (Q3GA)는 신경 보호 효과가 있는 것으로 보고 되어 왔다. 본 연구에서는 SH-SY5Y 세포에서 1-methyl-4-phenyl pyridinium ($MPP^+$)에 의해 유도된 신경 독성에 대한 Q3GA의 신경 보호 효과와 그 분자 조절 기전을 조사하였다. Q3GA는 $MPP^+$에 의해 유도된 세포 사멸을 유의적으로 감소시켰으며 PARP 절단을 감소시켰다. 또한, Bax/Bcl-2 비율의 감소와 함께 $MPP^+$에 의해 증가된 세포 내 ROS를 감소시켰다. Q3GA는 $MPP^+$에 의해 감소된 Akt와 CREB의 인산화를 유의적으로 회복시켰지만, ERK에는 영향을 미치지 않았다. 이 결과는 Q3GA가 ROS 생산 억제와 Akt/CREB 신호 전달 경로를 통해 $MPP^+$ 에 의해 유도된 신경 독성을 억제시킬 수 있음을 시사한다. 본 연구는 Q3GA가 파킨슨병에 대한 예방제 또는 치료제로 개발될 수 있는 가능성을 제시한다.

• 생명과학회지
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• 제29권1호
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• pp.129-134
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• 2019

이전 연구에서 전통주 주박 ethyl acetate 분획물로부터 11개의 순수물질을 분리 동정하였다. 11개의 순수물질은 caffeic acid, coumaric acid, D-mannitol, ferulic acid, hesperetin, hesperidin, naringenin, naringin, sinapic acid, syringic acid, 그리고 vanilic acid로 동정되었다. 이번 연구에서는 그들의 항염증 활성을 연구하기 위하여 LPS로 활성화된 RAW 264.7 세포에서 nitric oxide (NO) 생산을 측정하였다. 11개의 순수물질 중 hesperetin과 naringenin이 가장 높은 NO 생성 억제를 보여주었다. 또한, hesperetin은 세포 생존율에 영향 없이 농도의존적으로 NO 생산을 저해하였다. 그리고, hesperetin은 농도의존적으로 염증유전자인 iNOS의 발현을 농도의존적으로 억제한 반면, COX-2 단백질의 발현에는 영향을 주지 않았다. 게다가, hesperetin은 p38 MAPK와 ERK1/2의 인산화를 억제한 반면 JNK의 인산화에는 영향을 주지 못했다. 이러한 결과는 hesperetin은 항염증 활성을 가지며, 이러한 항염증 활성은 p38 MAPK와 ERK1/2 경로를 억제함으로써 일어난다는 것을 나타낸다.

• 대한한방부인과학회지
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• 제32권2호
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• pp.1-17
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• 2019

Objectives: This study was performed to identify the anti-inflammatory effects of Salvia miltiorrhizae radix Water extract (SMW) on lipopolysaccharide (LPS) induced inflammation. Methods: RAW 264.7 cells were treated with 500 ng/ml of LPS. SMW (0.1, 0.25, 0.5 mg/ml) was treated 1 h prior to LPS. Cell viability was measured by MTT assay. Levels of nitric oxide (NO) were measured with Griess reagent and pro-inflammatory cytokines were measured by enzyme-linked immunosorbent assay (ELISA) and real-time polymerase chain reaction (PCR). We also examined molecular mechanisms such as mitogen-activated protein kinases (MAPKs) and nuclear factor-kappa B ($NF-{\kappa}B$) activation by western blot. In addition, we observed mice survival rate after LPS and examined their cytokine levels of serum and liver tissue. Results: SMW itself did not have cytotoxic effects in RAW 264.7 cells less than 0.5 mg/ml. SMW treatment inhibited the production of NO, and interleukin $(IL)-1{\beta}$ which is pro-inflammatory cytokine. And SMW treatment inhibited the LPS-induced activation of MAPKs such as extracellular signal-regulated kinase1/2 (ERK1/2), p38 kinases (p38), c-Jun NH2-terminal kinase (JNK) and $NF-{\kappa}B$. In addition, it also showed reducing the level of $IL-1{\beta}$ on the serum and liver tissue of mice. Also, death of LPS-induced mice was inhibited by SMW. Conclusions: The result suggests that treatment of SMW could reduce the LPS-induced inflammation. Thereby, SMW could be used as a protective agent against inflammation. Also, this study could give a clinical basis that SMW could be a drug or agent to prevent inflammatory diseases.

• Journal of Applied Biological Chemistry
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• 제63권4호
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• pp.421-428
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• 2020

본 연구는 황기와 지치 복합물인 ALM16이 lipopolysaccharide 처리에 의해 자극된 RAW264.7 대식세포의 염증반응에 미치는 영향에 대하여 조사하였다. ALM16은 RAW264.7 대식세포에 대하여 최대 200 ㎍/mL의 농도까지 독성은 보이지 않았다. 항염증 활성을 검정하기 위해 nitric oxide (NO), prostaglandin E2 (PGE2) 및 pro-inflammatory cytokines 생성량을 측정한 결과, ALM16은 각각의 생성량을 농도의존적으로 감소시켰다. 또한 ALM16은 NO와 PGE2 생성에 관여하는 inducible nitric oxide synthase (iNOS)와 cyclooxygenase-2 (COX-2)의 단백질 발현을 억제하였다. 한편, 항염증 활성 조절 기전을 확인하기 위하여 NK-κB의 핵으로의 이동과 DNA-binding activity 및 MAPK 신호전달 경로에 대한 ALM16의 영향을 확인한 결과, ALM16은 NF-κB의 핵으로 이동과 DNA-binding activity를 유의적으로 억제하였으며, JNK와 ERK 특이적으로 인산화를 억제함으로써 MAPK 신호전달 경로 활성을 억제하였다. 이러한 결과를 종합하여 볼 때 ALM16이 MAPK와 NF-κB의 신호전달 경로 억제를 통한 iNOS와 COX-2의 발현을 조절하고, 이로 인하여 NO, PGE2 및 pro-inflammatory cytokines의 생성이 감소하여 염증 반응을 조절하는 능력이 있는 것으로 판단된다.

• Biomolecules & Therapeutics
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• 제29권2호
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• pp.211-219
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• 2021

Alopecia is a distressing condition caused by the dysregulation of anagen, catagen, and telogen in the hair cycle. Dermal papilla cells (DPCs) regulate the hair cycle and play important roles in hair growth and regeneration. Myristoleic acid (MA) increases Wnt reporter activity in DPCs. However, the action mechanisms of MA on the stimulation of anagen signaling in DPCs is not known. In this study, we evaluated the effects of MA on anagen-activating signaling pathways in DPCs. MA significantly increased DPC proliferation and stimulated the G2/M phase, accompanied by increasing cyclin A, Cdc2, and cyclin B1. To elucidate the mechanism by which MA promotes DPC proliferation, we evaluated the effect of MA on autophagy and intracellular pathways. MA induced autophagosome formation by decreasing the levels of the phospho-mammalian target of rapamycin (phospho-mTOR) and increasing autophagy-related 7 (Atg7) and microtubule-associated protein 1A/1B-light chain 3II (LC3II). MA also increased the phosphorylation levels of Wnt/β-catenin proteins, such as GSK3β (Ser9) and β-catenin (Ser552 and Ser675). Treatment with XAV939, an inhibitor of the Wnt/β-catenin pathway, attenuated the MA-induced increase in β-catenin nuclear translocation. Moreover, XAV939 reduced MA-induced effects on cell cycle progression, autophagy, and DPC proliferation. On the other hand, MA increased the levels of phospho (Thr202/Tyr204)-extracellular signal regulated kinases (ERK). MA-induced ERK phosphorylation led to changes in the expression levels of Cdc2, Atg7 and LC3II, as well as DPC proliferation. Our results suggest that MA promotes anagen signaling via autophagy and cell cycle progression by activating the Wnt/β-catenin and ERK pathways in DPCs.

• Animal Bioscience
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• 제35권1호
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• pp.115-125
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• 2022

Objective: Intramuscular fat (IMF) is a critical economic indicator of pork quality. Studies on IMF among different pig breeds have been performed via high-throughput sequencing, but comparisons within the same pig breed remain unreported. Methods: This study was performed to explore the gene profile and identify candidate long noncoding RNA (lncRNAs) and mRNAs associated with IMF deposition among Laiwu pigs with different IMF contents. Based on the longissimus dorsi muscle IMF content, eight pigs from the same breed and management were selected and divided into two groups: a high IMF (>12%, H) and low IMF group (<5%, L). Whole-transcriptome sequencing was performed to explore the differentially expressed (DE) genes between these two groups. Results: The IMF content varied greatly among Laiwu pig individuals (2.17% to 13.93%). Seventeen DE lncRNAs (11 upregulated and 6 downregulated) and 180 mRNAs (112 upregulated and 68 downregulated) were found. Gene Ontology analysis indicated that the following biological processes played an important role in IMF deposition: fatty acid and lipid biosynthetic processes; the extracellular signal-regulated kinase cascade; and white fat cell differentiation. In addition, the peroxisome proliferator-activated receptor, phosphatidylinositol-3-kinase-protein kinase B, and mammalian target of rapamycin pathways were enriched in the pathway analysis. Intersection analysis of the target genes of DE lncRNAs and mRNAs revealed seven candidate genes associated with IMF accumulation. Five DE lncRNAs and 20 DE mRNAs based on the pig quantitative trait locus database were identified and shown to be related to fat deposition. The expression of five DE lncRNAs and mRNAs was verified by quantitative real time polymerase chain reaction (qRT-PCR). The results of qRT-PCR and RNA-sequencing were consistent. Conclusion: These results demonstrated that the different IMF contents among pig individuals may be due to the DE lncRNAs and mRNAs associated with lipid droplets and fat deposition.

• Journal of Applied Biological Chemistry
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• 제65권2호
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• pp.113-120
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• 2022

구슬말(Nostoc commune Vaucher ex Bornet & Flahault)은 이형세포를 갖는 특징으로 다른 목들과 구분되는 남조류의 일종으로 극지방에서 열대지역까지 광범위한 지역에 분포하며 단세포가 연결되어 형성된 수많은 trichome들이 점액질에 둘러 쌓인 형태로 커다란 군체를 형성한다. 주로 토양, 암반, 잔디 위 등에 서식한다고 알려져 있으나 흔히 관찰되지 않기 때문에 현재 연구가 거의 없는 실정이다. 이에 본 연구에서는 토양 남조류인 N. commune HCW0811을 분리 및 동정하였으며 항염증 활성을 조사 하고자 하였다. 그 결과 N. commune HCW0811는 LPS로 유도된 RAW 264.7세포에서 80%이상의 세포 생존율을 나타내었으며 NO, PGE2 및 TNF-α, IL-6, IL-1β의 생성을 효과적으로 억제하였다. 또한 western blot assay를 통해 iNOS, COX-2 및 MAP kinase (p38, ERK1/2, JNK)와 NF-κB 세포내 신호전달 경로에서의 단백질 발현을 조사한 결과 이들의 발현이 유의하게 억제됨을 확인하였다. 본 연구에서는 이러한 결과를 근거하여 HCW0811가 다양한 염증 인자를 표적으로 하는 피부 면역 질환을 포함한 염증성 질환의 예방과 치료를 위한 항염증 기능성 화장품 및 식품소재로의 개발가능성을 제시한다.

• 대한한방부인과학회지
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• 제34권3호
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• pp.29-48
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• 2021

Objectives: Banchong-san (BCS) is a herbal formula composed of 13 korean medicinal herbs and is traditionally used to treat inflammatory diseases and pain. The object of this study was to research the antioxidant, anti-inflammatory, antipruritic and antimicrobial effects of the BCS in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. Methods: In this experiment, effects of BCS on the following four were measured as follows: (1) Anti-oxidative effects were evaluated by 1,1-diphenyl-2-picryl-hydrazyl (DPPH) Radical scavenging activity, 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) Radical scavenging activity. (2) Anti-inflammatory effects were evaluated by the production amount of Reactive oxygen species (ROS), Nitric oxide (NO), Interleukin-1β (IL-1β), Interleukin 6 (IL-6), tumor necrosis factor-α (TNF-α), Prostaglandin E2 (PGE₂), inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2)(the previous two are "mRNA"), extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), p38 mitogen-activated protein kinases (p38), inhibitor of nuclear factor kappa B (IκBα), nuclear factor kappa B (NF-κB) (the previous five are "Protein") in LPS-Stimulated RAW 264.7 cells. (3)Antipruritic effects were evaluated by the production amount of histamine, Leukotriene B4 (LTB4), LeukotrieneC4 (LTC4) Levels in phorbol 12-myristate 13-acetate(PMA)/ionomycin-stimulated MC/9 mast cell. (4) Anti-microbial effects were evaluated by the growth suppression of Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa and Aspergillus niger. Results: The following results were obtained through each measurement: (1) DPPH Radical Scavenging Activity, ABTS Radical Scavenging Activity evoked a significant concentration-dependent increase. (2) ROS, NO, IL-1β, IL-6, TNF-α, PGE₂ production amount, iNOS, COX-2 mRNA expression were significantly reduced in the BCS extraction group compared with the control group and significantly decreased the amount of ERK, JNK, p38, NF-κB Protein expression. The amount of IκB-α Protein Expression have increased significantly. (3) The amounts of histamine, LTB4, LTC4 were significantly decreased. (4) The antibacterial efficacy, BCS inhibited the growth of Escherichia coli, Pseudomonas aeruginosa at concentrations of 5 ㎍/ml, but did not suppress the growth of staphylococcus aureus and aspergillus niger. Conclusions: The experimental results show that BCS has anti-oxidant, anti-inflammatory, antipruritic and antimicrobial properties.

• BMB Reports
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• 제54권5호
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• pp.266-271
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• 2021

Estrogen-related receptor γ (ERRγ), a member of the orphan nuclear receptor family, is a key mediator in cellular metabolic processes and energy homeostasis. Therefore, ERRγ has become an attractive target for treating diverse metabolic disorders. We recently reported that ERRγ acts as a negative regulator of osteoclastogenesis induced by receptor activator of nuclear factor-κB ligand (RANKL). In the present study, we explored the effects of an ERRγ-specific modulator, GSK5182, on ERRγ-regulated osteoclast differentiation and survival. Interestingly, GSK5182 increased ERRγ protein levels much as does GSK4716, which is an ERRγ agonist. GSK5182 inhibited osteoclast generation from bone-marrow-derived macrophages without affecting cytotoxicity. GSK5182 also attenuated RANKL-mediated expression of cFos and nuclear factor of activated T-cells cytoplasmic 1 (NFATc1), pivotal transcription factors for osteoclastogenesis. Arrested osteoclast differentiation was associated with reduced RANK expression, but not with the M-CSF receptor, c-Fms. GSK5182 strongly blocked the phosphorylation of IκBα, c-Jun N-terminal kinase, and extracellular signal-regulated kinase in response to RANKL. GSK5182 also suppressed NF-κB promoter activity in a dose-dependent manner. In addition to osteoclastogenesis, GSK5182 accelerated osteoclast apoptosis by caspase-3 activation. Together, these results suggest that GSK5182, a synthetic ERRγ modulator, may have potential in treating disorders related to bone resorption.

• 대한본초학회지
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• 제36권3호
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• pp.47-53
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• 2021

Objectives : Currently, the alopecia is one of the most emotionally stressful syndromes in human life. Human hair dermal papilla cells (HDPCs) play an essential role in controlling hair growth and in regulating hair cycle. We performed MTT assay, cell cycle, and western blot to determine the effects of essential oil from Coicis Semen (ECS) on hair growth in HDPCs. Methods : We monitored cell proliferations by MTT assay in HDPCs. After setting up the safe and effective concentration range to be treated ECS, cell cycle analysis was performed using flow cytometry. Also, the protein expression of hair growth-related factors such as insulin like growth factor-1 (IGF-1), Wnt, extracellular signal-regulated kinase (ERK), serine/threonine-specific protein kinase (Akt) in HDPCs was determined by western blot. Results : As results, cell proliferation was increased in ECS group compared to dimethyl sulfoxide (DMSO) group and minoxidil (MNXD) group. Cell number of ECS group was more decrease in sub G1 phase than cell number of DMSO group. Also, cell number of ECS group increased compared to cell number of DMSO group in G1 phase. Protein expression of ECS group was higher than protein expression of DMSO group on related hair growth factors (IGF-1, Wnt, ERK, Akt). Conclusion : As mentioned above, ECS increased cell proliferation and the protein expression of IGF-1, Wnt, ERK, and Akt. These results suggest that ECS could be used as a potential material for the treatment of alopecia by increasing the proliferation of HDPCs.