• The Korean Journal of Physiology and Pharmacology
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• 제13권3호
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• pp.229-239
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• 2009

The aim of the present study was to examine the effect of provinol, which is a mixture of polyphenolic compounds from red wine, on the secretion of catecholamines (CA) from isolated perfused rat adrenal medulla, and to elucidate its mechanism of action. Provinol (0.3 ${\sim}$ 3 ${\mu}g/ml$) perfused into an adrenal vein for 90 min dose- and time-dependently inhibited the CA secretory responses evoked by ACh (5.32 mM), high $K^+$ (a direct membrane-depolarizer, 56 mM), DMPP (a selective neuronal nicotinic $N_N$ receptor agonist, 100 ${\mu}M$) and McN-A-343 (a selective muscarinic $M_1$ receptor agonist, 100 ${\mu}M$). Provinol itself did not affect basal CA secretion. Also, in the presence of provinol (1 ${\mu}g/ml$), the secretory responses of CA evoked by Bay-K-8644 (a voltage-dependent L-type dihydropyridine $Ca^{2+}$ channel activator, 10 ${\mu}M$), cyclopiazonic acid (a cytoplasmic $Ca^{2+}$-ATPase inhibitor, 10 ${\mu}M$) and veratridine (an activator of voltage-dependent $Na^+$ channels, 10 ${\mu}M$) were significantly reduced. Interestingly, in the simultaneous presence of provinol (1 ${\mu}g/ml$) plus L-NAME (a selective inhibitor of NO synthase, 30 ${\mu}M$), the CA secretory responses evoked by ACh, high $K^+$, DMPP, McN-A-343, Bay-K-8644 and cyclpiazonic acid recovered to the considerable extent of the corresponding control secretion in comparison with the inhibition of provinol-treatment alone. Under the same condition, the level of NO released from adrenal medulla after the treatment of provinol (3 ${\mu}g/ml$) was greatly elevated in comparison to its basal release. Taken together, these data demonstrate that provinol inhibits the CA secretory responses evoked by stimulation of cholinergic (both muscarinic and nicotinic) receptors as well as by direct membrane-depolarization from the perfused rat adrenal medulla. This inhibitory effect of provinol seems to be exerted by inhibiting the influx of both calcium and sodium into the rat adrenal medullary cells along with the blockade of $Ca^{2+}$ release from the cytoplasmic calcium store at least partly through the increased NO production due to the activation of nitric oxide synthase.

• 인지과학
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• 제12권4호
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• pp.49-56
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• 2001

입체시 계산 모형들은 국지적으로 대응 가능한 여러 대상들이 있는 경우 가설적인 부등 탐지기들간의 상호 작용을 통해 대응될 대상을 결정한다. 이 연구에서는 인간의 양안시 기제도 입체시 모형들의 가정과 유사하게 부등 탐지기들의 활성화 수준에 따라 대응될 대상을 선택하는 지를 알아보기 위해서 깊이가 다른 두 표면을 가지는 무선점 패턴을 입체 그림으로 사용하였다. 응시 표면과 같은 깊이에 놓여 있는 표면을 구성하는 점들은 무선적으로 선택되었으며(원 무선 패턴) 이 표면보다 앞에 놓여있는 표면을 구성할 점들은 원 무선 패턴의 각 점들과 다음과 같은 방식으로 짝지어졌다. 원 무선 패턴을 구성하는 점들의 위치를 발견하여 왼쪽 입체 그림에서는 무선 점들의 왼쪽 3분에 오른쪽 입체 그림에서는 무선 점들의 오른쪽 3분에 위치하도록 하여 6 분의 깊이에 해당하는 또 다른 표면을 만들었다. 원 무선점 패턴의 자기를 고정시킨 채 새로운 패턴의 밝기를 변화시키면 가설적인 탐지기들의 상대적인 활성화 수준이 서로 달라질 것을 추정할 수 있다. 상대적인 활성화 수준에 따라 대응될 대상이 변화되는 지를 알아보기 위해서 모든 점들의 위치는 고정시킨 채 두 표면을 구성하는 점들간의 상대적인 밝기를 변화시켜가면서 각 밝기 수준에서 두 표면의 깊이를 측정하였다. 쌍을 이루는 점들간의 밝기 차가 30% 이내인 조건에서는 3 분의 부등에 해당되는 하나의 표면이 지각되었지만, 두 점의 밝기가 타이가 날수록 이 표면 이외의 다른 표면과 더불어 모두 두개의 표면이 지각되었다. 특히 새로운 표면의 깊이는 무선점 패턴을 구성하는 점들과 추가된 점들의 상대적 밝기에 따라 달라겠다. 추가된 점들이 밝은 조건에서는 이 점들의 부등에 해당되는 깊이를 갖는 표면이 원 무선점 패턴이 밝은 조건에서는 무선점 패턴의 부등에 해당되는 깊이를 가진 표면이 추가적으로 지각되었다. 이러한 결과는 보다 활성화 된 탐지기들이 약한 탐지기들을 억제하여 있음을 시사한다.

• Asian Pacific Journal of Cancer Prevention
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• 제16권6호
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• pp.2251-2256
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• 2015

To study effects of cellular FLICE (FADD-like IL-$1{\beta}$-converting enzyme)-inhibitory protein (c-FLIP) inhibition by RNA interference (RNAi) on sensitivity of U2OS cells to tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL)-induced apoptosis, plasmid pSUPER-c-FLIP-siRNA was constructed and then transfected into U2OS cells. A stable transfection cell clone U2OS/pSUPER-c-FLIP-siRNA was screened from the c-FLIP-siRNA transfected cells. RT-PCR and Western blotting were applied to measure the expression of c-FLIP at the levels of mRNA and protein. The results indicated that the expression of c-FLIP was significantly suppressed by the c-FLIP-siRNA in the cloned U2OS/pSUPER-c-FLIP-siRNA as compared with the control cells of U2OS/pSUPER. The cloned cell line of U2OS/pSUPER-c-FLIP-siRNA was further examined for TRAILinduced cell death and apoptosis in the presence of a pan-antagonist of inhibitor of apoptosis proteins (IAPs) AT406, with or without 4 hrs pretreatment with rocaglamide, an inhibitor of c-FLIP biosynthesis, for 24 hrs. Cell death effects and apoptosis were measured by the methods of MTT assay with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and flow cytometry, respectively. The results indicated that TRAIL-induced cell death in U2OS/pSUPER-c-FLIP-siRNA was increased compared with control cells U2OS/pSUPER in the presence or absence of AT406. Flow cytometry indicated that TRAIL-induced cell death effects proceeded through cell apoptosis pathway. However, in the presence of rocaglamide, cell death or apoptotic effects of TRAIL were similar and profound in both cell lines, suggesting that the mechanism of action for both c-FLIP-siRNA and rocaglamide was identical. We conclude that the inhibition of c-FLIP by either c-FLIP-siRNA or rocaglamide can enhance the sensitivity of U2OS to TRAIL-induced apopotosis, suggesting that inhibition of c-FLIP is a good target for anti-cancer therapy.

• Asian Pacific Journal of Cancer Prevention
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• 제15권16호
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• pp.6829-6835
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• 2014

Chemotherapy is the primary therapy for malignant lymphoma (ML). However, the clinical outcome is still far from satisfactory. Consequently, an understanding of the mechanism of modulating cancer cell invasion, migration and metastasis is important for the development of more effective chemotherapeutic agents. FNC, 2'-deoxy-2'-${\beta}$-fluoro-4'-azidocytidine, a novel cytidine analogue, has demonstrated significantly inhibitory effects on proliferation of several non-Hodgkin lymphoma (NHL) cell lines. A previous study indicated that FNC effectively inhibited the growth of Raji and JeKo-1 cells in dose-time dependent effects with $IC_{50}$ values of $0.2{\mu}M$ and $0.097{\mu}M$, respectively. This study was focused on investigating the anti-invasive properties of FNC on NHL cells and its potential mechanisms of action. Cell adhesion and transwell chamber assays were utilized to investigate the anti-invasive effects of FNC on Raji and JeKo-1 cells. Real-time PCR and Western blotting were employed to qualify the expression of ${\beta}$-catenin, the glycogen synthase kinase-3 beta (GSK-$3{\beta}$), E-cadherin vascular endothelial growth factor (VEGF), matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9). The results revealed that FNC remarkably inhibited the adhesion, migration and invasion of two human aggressive non-Hodgkin lymphoma cell lines in a dose dependent manner. Furthermore, ${\beta}$-catenin, MMP-2, MMP-9, VEGF mRNA and protein levels were decreased after FNC treatment, while GSK-$3{\beta}$ and E-cadherin increased. Our studies thus provide evidence and a rationale that FNC may offer an effective chemotherapeutic agent by regulating the invasion and metastasis of aggressive non-Hodgkin lymphoma via inhibition of the Wnt/${\beta}$-catenin signaling pathway.

• Asian Pacific Journal of Cancer Prevention
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• 제14권12호
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• pp.7179-7186
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• 2013

Background and Aims: Scutellaria is one of the most popular traditional Chinese herbal remedies against various human diseases, including cancer. In this study, we examined the active effects of Scutellaria extract and its main flavonoid constituents on the proportion of side population cells within human multiple myeloma cell line RPMI8226 in vitro and explored the potential molecular mechanisms involved. Materials and Methods: The contents of flavonoids in ethanolic extract of Scutellaria baicalensis Georgi were determined using high performance liquid chromatography. The antiproliferative effect of the ethanolic extract on RPMI-8226 was determined by CCK assay. Apoptosis was measured by annexin combining with propidium iodide in a flow cytometer. Cell cycle analysis was performed by propidium iodide staining in combination with flow cytometry analysis. Hoechst 33342 exclusion assay was used for the identification of side population within RPMI8226 cells. The expression of ABCG2 protein was assessed by Western blotting assay. Results: The content of major flavonoids constitutents of Scutellaria extract was baicalin (10.2%), wogonoside (2.50%), baicalein (2.29%), and wogonin (0.99%), respectively. The crude Scutellaria extract did not show significant anti-proliferative effect, apoptosis induction and cell cycle arrest in RPMI-8226 within the concentrations of $1-75{\mu}g/mL$. However, the ethanolic extract, baicalein, wogonin and baicalin reduced the side population cells in RPMI-8226, and data showed that baicalein and wogonin had stronger inhibitory effects. Correspondingly, they also exhibited significant effects on decreasing the expression level of ABCG2 protein in RPMI-8226 in vitro. Conclusions: Our results for the first time demonstrated a novel mechanism of action for Scutellaria extract and its main active flavonoids, namely targeting SP cells by modulating the expression of ABCG2 protein. This study provides an insight for new therapeutic strategies targeting cancer stem cells of multiple myeloma.

• The Korean Journal of Physiology and Pharmacology
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• 제9권6호
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• pp.333-339
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• 2005

B/K protein is a novel protein containing double C2-like domains. We examined the specific signaling pathway that regulates the transcription of B/K in PC12 cells. When the cells were treated with forskolin ($50{\mu}M$), B/K mRNA and protein levels were time-dependently decreased, reaching the lowest level at 3 or 4 hr, and thereafter returning to the control level. Chemicals such as dibutyryl-cAMP, cellpermeable cyclic AMP (cAMP) analogue and CGS21680, adenosine receptor $A_{2A}$ agonist, also repressed the B/K transcription. However, 1,9-dideoxyforskolin did not show inhibitory effect on B/K transcription, suggesting direct involvement of cAMP in the forskolin-induced inhibition of B/K transcription. Effect of forskolin, dibutyryl cAMP and CGS21680 was significantly reduced in PKA-deficient PC12 cell line (PC12-123.7). One cAMP-response element (CRE)-like sequence (B/K CLS) was found in the promoter region of B/K DNA, and electrophoretic mobility shift assay indicated its binding to CREM and CREB. Forskolin significantly suppressed the promoter activity in CHO-K1 cells transfected with the constructs containing B/K CLS, but not with the construct in which B/K CLS was mutated (AC：TG). Taken together, we suggest that the transcription of B/K gene in PC12 cells may be regulated by PKA-dependent mechanism.

• 한국동물위생학회지
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• 제32권4호
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• pp.353-359
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• 2009

Interest in marine organisms as potential sources of bioactive agents has increased in recent years. The red seaweed, Callophyllis (C.) japonica, is abundant in the coastal regions of Jeju Island in South Korea. A previous study shows that C. japonica extracts have antioxidant activity and radioprotective effects. In this study, an methanol extract of C. japonica was tested whether it has antibacterial effects against the bacteria from swine. In vitro antibacterial activities of the crude extracts prepared from the C. japonica using 80 % methanol were tested for inhibitory activity against the Escherichia (E.) coli (S175), Enterococcus (E.) faecium (ATCC 51558), Salmonella (S.) Typhimurium and Staphylo-coccus (S.) aureus (ATCC 25923) by using broth dilution method. All organisms were incubated in brain heart infusion medium containing 1% extract at 0, 4, 8, 12 and 24 hrs. The 3 days-old piglets were fed an experimental diet supplemented with 1% C. japonica for 1 week. And the change of the coliform bacteria in feces were examined after supplement of C. japonica for 1 week. When the inocula containing $10^2{\sim}10^3$CFU/ml of each organism were used the extracts of C. japonica showed various degrees of antibacterial effects on all bacteria tested. The CFU value ($6.3\times10^8$CFU/ml) of C. japonica for E. coli was decreased 30% compared with vehicle controls ($9.0\times10^8$CFU/ml) after 8 hrs incubation. The proliferation rate of E. faecium was inhibited about 68% at 4 hrs, 81% at 8 hrs and 76% at 12 hrs after incubation, respectively. The proliferation rate of S. Typhimurium was inhibited about 96% at 4 hrs, 90% at 8 hrs and 72% at 12 hrs after incubation with extracts of C. japonica. The proliferation rate of S. aureus was inhibited more than 90% each time courses. Conclusively, a red seaweed extract of C. japonica was found to be effective against a number of gram negative and gram positive bacteria such as E. coli, E. faecium, S. Typhimurium, and S. aureus. The number of coliform bacteria was increased in the 1% C. japonica-treated group, as compared to those of controls. This result suggests that C. japonica extracts be added as an effective natural antibacterial agent. The precise mechanism of antibacterial effects and its application on swine industry remains to be further studied.

• 한국잡초학회지
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• 제18권4호
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• pp.333-340
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• 1998

Primisulfuron 또는 Imazethapyr와 Cytochrome P-450 저해제인 PBO의 혼합처리가 옥수수의 생장 및 ALS효소 활성에 미치는 영향을 조사한 결과는 다음과 같다. 1. Pioneer 3751 IR은 Primisulfuron과 Imazethapyr에 Imazethapyr에 높은 내성을 보인 반면, Pioneer 3751과 찰옥 2호는 감수성을 보였다. 2. Pioneer 3751 IR은 Primisulfuron과 PBO혼합처리에서 높은 저항성을 나타내어 PBO혼합처리에 따른 영향은 적었으나 Pioneer 3751,수원 118호와 찰옥 2호는 PBO혼합처리에 따라 생장 저해가 PBO무처리에 비해 높게 나타났다. 3. Pioneer 3751 IR의 생장은 Imazethapyr와 PBO 혼합처리에 따른 영향은 적었으며 Pioneer 3751과 수원 118호, 찰옥 2호는 생장저해가 심하였으나 PBO혼합처리 에 따른 약해 상승작용은 크지 않았다. 4. ALS효소 활성은 Prirnisulfuron은 $0.01{\mu}M$에서부터, 그리고 Imazethapyr는 $1{\mu}M$에서부터 Pioneer 3751 IR 품종을 제외한 3품종에서 현저하게 억제되었으며, Primisulfuron과 PBO혼합처리에 의하여 Pioneer 3751 IR과 Pioneer 3751에서는 활성저해가 더 크게 나타났고 수원 118호와 찰옥 2호에서는 차이가 없었으며, Imazethapyr와 PBO의 혼합처리에서는 4품종에서 모두 약간 억제되었다.

• 한국잡초학회지
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• 제14권4호
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• pp.280-290
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• 1994

작물(作物)과 잡초(雜草)에서 Oxyfluorfen과 Chlormethoxynil에 대한 선택성(選擇性)과 살초력(殺草力)을 비교(比較)하고자 발아실험(發芽實驗), 수경재배실험(水耕栽培實驗), 두가지 토양(土壤)에서의 발아전(發芽前) 토양처리실험(土壤處理實驗)을 실시(實施)하였으며, 처리부위(處理部位)와 시간(時間)을 다르게 하였을 때의 초종간(草種間) 흡수(吸收) 및 이행(移行)을 조사하고자 $^{14}C$-oxyfluorfen을 이용(利用)한 흡수이행실험(吸收移行實驗)을 실시(實施)하였다. 1. 발아실험(發芽實驗), 수경재배실험(水耕栽培實驗), 발아전(發芽前) 토양처리실험(土壤處理實驗)을 통하여 볼 때 벼, 옥수수에서는 공시제초제(供試除草劑) 농도(濃度)가 증가(增加)되면서 약간 생장(生長)이 억제(抑制)되는 경향(傾向)을 나타냈으나 감수성(感受性) 초종(草種)인 참소리쟁이, 바랭이에서는 농도(濃度)가 높아짐에 따라 살초력(殺草力)이 증대(增大)되어 생장억제(生長抑制)가 심하였으며 전반적(全般的)으로 Oxytluorfen처리(處理)는 Chlormethoxynil처리(處理)보다 생장억제(生長抑制)가 심하였다. 2. 발아기간(發芽期間)동안의 생육억제(生育抑制) 효과(效果)를 나타낸 것을 보면 발아과정(發芽過程) 때보다 발아후(發芽後) 생육과정(生育過程)에서 많은 것으로 사료(思料)되며, 벼와 옥수수에서 가장 둔감(鈍感)한 반응(反應)을, 피에서는 비교적 둔감(鈍感)한 반응(反應)을, 참소리쟁이와 바랭이에서는 가장 민감(敏感)한 반응(反應)을 나타내었다. 3. 수경재배실험(水耕栽培實驗) 결과(結果), 2엽기(葉期)에서의 처리(處理)가 4엽기(葉期)에서의 처리(處理)보다 생장억제(生長抑制)가 더 심하게 나타났으며 지상부(地上部)보다 지하부(地下部)가 많은 생장억제(生長抑制)를 나타내었다. 4. 발아전(發芽前) 토양처리실험(土壤處理實驗) 결과(結果), 유기물(有機物), C.E.C. 및 점토성분(粘土成分)이 적은 사양토(砂壤土)에서 생장억제(生長抑制)가 심하였으며 초종간(草種間)에는 참소리쟁이와 바랭이에서 많이 나타났다. 5. $^{14}C$-oxytluorfen의 흡수(吸收)는 작물(作物)과 잡초(雜草)에서 모두 잘 되였으나 이행은 모든 초종(草種)에서 거의 되지 않는 경향이었으나 시간(時間)이 경과(經過)함에 따라 감수성(感受性) 초종(草種)인 참소리쟁이와 바랭이에서 흡수량(吸收量)이 많아졌으며 작물(作物)과 잡초간(雜草間) 선택성(選擇性)은 주로 흡수(吸收)된 양(量)에 따라 결정된다고 생각한다.

• 한국자원식물학회지
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• 제33권5호
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• pp.428-435
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• 2020

본 연구에서 상동나무 가지 추출물(STB-E100)은 대장암 세포에서 세포사멸을 유도하여 세포생육을 억제하였다. 또한 IκB-α 인산화를 통한 IκB-α 단백질 분해를 유도하며 이로 인해 P65 핵내 전이를 유도하여 NF-κB 신호전달을 활성화시킨다. NF-κB 신호전달 활성화는 GSK3β 활성화를 통해 P65 핵내 전이를 유도에 의한 것이지만 IκB-α분해는 GSK3β 의존성이 아니다. 상동나무 가지 추출물은 이러한 신호전달 활성화를 통해 세포사멸을 유도하여 대장암의 세포생육을 억제한다. 본 결과를 바탕으로 상동나무 가지가 암 예방 및 치료를 목적으로 한 표적 요법에서 항암제 개발의 잠재적 활용 소재로서 이용 가능하다고 사료된다. 그러나 대장암 세포에서 상동나무 가지 추출물에 의해 유도된 NF-κB 신호전달 작용기전을 좀더 구체적으로 구명할 필요가 있고 대장암에 대한 세포사멸과 작용기전의 정확한 관련성을 조사하기 위해 추가적인 연구가 필요하다.