• Journal of Ginseng Research
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• 제37권4호
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• pp.401-412
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• 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.

• 생명과학회지
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• 제19권6호
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• pp.836-838
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• 2009

뇌졸중 치료에 사용되는 tPA는 탁월한 혈전 용해 효과를 보이고 있어 혈액의 흐름을 용이하게 하는데 중요한 역할을 한다. 그러나 tPA 치료법은 매우 짧은 시간 내에 사용해야하는 단점과 출혈, 부종과 같은 여러 가지 부작용이 수반될 수 있기 때문에 매우 제한적이다. 이전의 실험 결과에 따르면 tPA의 이러한 양면적인 현상은 MMP의 활성 조절과 관련이 있는 것으로 보고되어 있으나 세포시스템을 활용한 이들의 직접적인 효과나 조절 기전에 대한 연구는 거의 알려져 있지 않다. 본 연구는 임상에서 사용되는 tPA의 부정적인 효과를 극복하기 위한 방안을 모색하고자 tPA와 MMP 활성과의 조절 기전을 살펴보았다. 랫트의 뇌로부터 추출한 신경세포에서 tPA의 처리는 MMP의 발현을 촉진시켰고 저산소상태에서 tPA에 의한 MMP활성 증가가 더욱 가속화되었으며 JNK 신호전달 경로를 통해 조절되는 것을 확인하였다.

• 한국작물학회지
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• 제40권2호
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• pp.133-141
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• 1995

약용 식물의 추출액이 자가영양배양세포의 광합성전자전달계에 미치는 영향을 조사하기 위해 9종의 약용식물 추출액으로부터 종자발아, PA세포의 엽록소 억제정도, DCIP의 환원율, 세포 생존율, 광계 I의 전자전달활성, 단백질에 미치는 영향을 조사한 결과는 다음과 같다. 1. 식물의 추출액을 농도별로 처리하였을 때 10％ 처리시 전 식물체에서 상추의 발아억제 현상을 나타내었고, 특히 백두옹과 초오 추출물 10％ 처리시는 100％ 억제를 나타내었다. 2. 백두옹의 증류수 및 MeOH 추출액을 PA세포에 처리한 경우 엽록소의 생성을 100％ 억제 하 였다. 이는 광합성 전자전달 저해제로 알려진 DCMU 10-3M 처리와 동일한 억제 효과였다. 3. PA세포에 추출물 처리시 백두옹이 힐반응 억제가 가장 컸으며, 세포 생존력은 가장 낮았다. 4. 광합성 산소발생은 반하, 독활, 백두옹, 만형자 추출액 처리시 14-77％ 억제되었고, 특히 PA 세포 2ml 반응액에 백두옹 추출물 60rl 처리시 50％ 산소발생 억제를 나타내었다. 5. 추출액을 PA 세포에 처리한 후 단백질을 추출하여 SDS-PAGE를 이용하여 조사한 결과 대조구에 비하여 백두옹 추출물 10％ 처리에서 14KD, 31KD, 41KD, 53KD, 73KD의 밴드가 나타나지 않았다.

• IMMUNE NETWORK
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• 제12권5호
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• pp.213-216
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• 2012

Our previous report showed that polyacetylene compound, 1-Heptadecene-11, 13-diyne-8, 9, 10-triol (PA) from the root of Cirsium japonicum var. ussuriense has anti-inflammatory activity. In this study we investigated the role of the PA as inhibitor of caspase-1, which converts prointerleukin-$1{\beta}$ (proIL-$1{\beta}$) to active IL-$1{\beta}$ and is activated by inflammasome involved in the inflammatory process. We tested the effect of PA on the production of pro-inflammatory cytokines, IL-$1{\beta}$ in murine macrophage cell line, RAW264.7. PA inhibited lipopolysaccharide (LPS)-induced IL-$1{\beta}$ production by macrophages at a dose dependent manner. PA also suppressed the activation of caspase-1. The mRNA level of ASC (apoptosis-associated spec-like protein containing a CARD), an important adaptor protein of inflammasome, was decreased in the PA treated group. Therefore our results suggest that the anti-inflammatory effect of PA is due to inhibit the caspase-1 activation.

• 한국발생생물학회지:발생과생식
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• 제3권1호
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• pp.29-38
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• 1999

자궁내막증은 흔한 부인과적 질병이며 여성 불임의 한 원인이 되나 그 발생 원인에 대하여서는 아직 논란의 여지가 많다. 최근 월경혈의 역류가 한 원인이며 자궁내막증 환자가 정상여성에서 보다 역류되는 월경혈의 양이 많거나 침습성이 강한 것이 자궁내막증의 발생 원인이 될 수 있다는 이론들이 소개되었다. 종양이나 자궁내막 조직의 침습이나 전이에는 세포막외 기질 및 기저막의 파괴가 일어나야 하는데 이 과정에 plasminogen activators (PAs)나 matrix metalloproteinase (MMPs)같은 proteolytic enzyme이 관여한다. 이에 자궁 내막증환자와 대조군의 자궁내막에서 PA나 MMP를 억제하는 plasminogen activator inhibitor-1 (PAI-1)나 tissue inhibitor of metalloproteinase (TIMP-3)의 mRNA 발현의 차이를 quantitative competitive RT PCR로 연구하였으며, 그 결과 자궁내막증 환자의 황체기 자궁내막에서는 정상 대조군 환자에서 보다 PAI-1과 TIMP-3 mRNA발현이 낮음을 알 수 있었다. 따라서 자궁내막증 환자의 자궁내막에서는 PA와 MMP의 활성도가 증가할 수 있으며 이 증가된 proteolytic activity로 인하여 역류된 자궁내 막 조직의 복강내 침습이 보다 쉽게 일어날 가능성이 있다.

• Journal of Microbiology and Biotechnology
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• 제10권2호
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• pp.233-237
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• 2000

As part of a continuing investigation to identify free radical scavengers from the fruit bodies of basidiomycetes, we isolated two p-terphenyl compounds, designated as PAl and PA2l, from cethanolic extract of the fruit body of Pasillus panuoides. The methanolic extract was processed by ethyl acetate extraction and silica gel column chromatography to yield two active fraction. PAl was obtained from one of the fractions through Sephadex LH-20 and silica gel column chromatographies and reverse-phase HPLC. The other fraction was purified by Sephadex LH-20 and reverse-phase column chromatographies to produce PA2. The compounds PA1 and PA2 were identified as leucomentin-4 and leucomentin-2, respectively, on the basis of various spectroscopic analyses. These compounds exhibited strong inhibitory activities against lipid peroxidation in rat liver microsomes with IC_{50}$ values of 0.10 and $0.06{\;}\mu\textrm{g}ml$, respectively.

• Journal of Microbiology and Biotechnology
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• 제20권8호
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• pp.1243-1250
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• 2010

A cDNA encoding a cysteine protease inhibitor (CPI) was isolated from the cDNA library of clamworm Perinereis aibuhitensis Grube. The deduced amino acid sequence analysis showed that the protein had 51%, 48%, and 48% identity with Zgc:153129 from Danio rerio, cystatin B from Theromyzon tessulatum, and the ChainA, stefin B tetramer from Homo sapiens, respectively. The gene was cloned into the intracellular expression vector pET-15b and expressed in Escherichia coli. The recombinant CPI (PA-CPI) was purified by affinity chromatography on Ni-charged resin and ion-exchange chromatography on DEAE-Sepharose FF. The relative molecular mass of PA-CPI was 16 kDa as deduced by SDS-PAGE. Activity analysis showed that the recombinant protein could inhibit the proteolytic activity of papain. A constitutive and secretive expression vector was also constructed, and the cDNA encoding CPI was subcloned into the vector for extracellular expression. Western blotting analysis results showed that the PA-CPI was secreted into the medium. Bioassay demonstrated that E. coli DH5${\alpha}$ harboring pUC18ompAcat-CPI showed a significant difference in mortality to the Asian longhorned beetle Anoplophora glabripennis compared with untransformed E. coli DH5${\alpha}$ and control.

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2004년도 제4회 발생공학 국제심포지움 및 학술대회
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• pp.108-108
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• 2004

• Reproductive and Developmental Biology
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• 제32권2호
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• pp.89-95
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• 2008

The present study was undertaken to identify changes of plasminogen activators (PAs) in porcine oviductal epithelial cells (POECs) during the estrous cycle classified with post-ovulatory stages (Post-Ov), early to mid-luteal stages (Early-mid L) and pre-ovulatory (Pre-Ov) stages. The urokinase-type plasminogen activator (uPA) was only observed on day 5 and day 7 of culture in the POECs on all the estrous cycles and gradually increased according to increasing culture times, but not Early-mid L. In POECs-conditioned medium, uPA, tissue-type (tPA) and tPA-PA inhibitor (tPA-PAI) activity were observed at all culture times during estrous cycles. The uPA activity of POECs-conditioned medium on Post-Ov stage were significantly (p<0.05) decreased during prolonged cultures. On the other hand, the tPA activity of POECs-conditioned medium at Post-Ov stage was significantly (p<0.05) higher on day 5 than compared to the other days. Although was higher on day 1 at Post-Ov stage, the tPA-PAI activity of POECs-conditioned medium was significantly (p<0.05) higher on day 7 at all stage than that of day 5 of the culture. Taken together, these results suggest that uPA, tPA and tPA-PAI are produced by POECs, and the variations of the PAs activity are regulated in the different stages of the estrous cycle.

• The Korean Journal of Physiology and Pharmacology
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• 제28권1호
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• pp.49-57
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• 2024

While arterial tone is generally determined by the phosphorylation of Ser¹⁹ in myosin light chain (p-MLC2), Thr¹⁸/Ser¹⁹ diphosphorylation of MLC2 (pp-MLC2) has been suggested to hinder the relaxation of smooth muscle. In a dual-wire myography of rodent pulmonary artery (PA) and mesenteric artery (MA), we noticed significantly slower relaxation in PA than in MA after 80 mM KCl-induced condition (80K-contraction). Thus, we investigated the MLC2 phosphorylation and the expression levels of its regulatory enzymes; soluble guanylate cyclase (sGC), Rho-A dependent kinase (ROCK) and myosin light chain phosphatase target regulatory subunit (MYPT1). Immunoblotting showed higher sGC-α and ROCK2 in PA than MA, while sGC-β and MYPT1 levels were higher in MA than in PA. Interestingly, the level of pp-MLC2 was higher in PA than in MA without stimulation. In the 80K-contraction state, the levels of p-MLC2 and pp-MLC2 were commonly increased. Treatment with the ROCK inhibitor (Y27632, 10 µM) reversed the higher pp-MLC2 in PA. In the myography study, pharmacological inhibition of sGC (ODQ, 10 µM) slowed relaxation during washout, which was more pronounced in PA than in MA. The simultaneous treatment of Y27632 and ODQ reversed the impaired relaxation in PA and MA. Although treatment of PA with Y27632 alone could increase the rate of relaxation, it was still slower than that of MA without Y27632 treatment. Taken together, we suggest that the higher ROCK and lower MYPT in PA would have induced the higher level of MLC2 phosphorylation, which is responsible for the characteristic slow relaxation in PA.