• Journal of Microbiology and Biotechnology
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• 제20권2호
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• pp.319-324
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• 2010

Phorbol ester extraction was carried out from Jatropha curcas seed cake, a by-product from the biodiesel fuel industry. Four repeated extractions from 5 g of J. curcas seed cake using 15 ml of 90% (v/v) ethanol and a shaking speed of 150 rpm gave the highest yield of phorbol esters. The ethanolic extract of J. curcas seed cake showed antifungal activities against important fungal phytopathogens: Fusarium oxysporum, Pythium aphanidermatum, Lasiodiplodia theobromae, Curvularia lunata, Fusarium semitectum, Colletotrichum capsid, and Colletotrichum gloeosporioides. The extract contained phorbol esters mainly responsible for antifungal activities. The extract could therefore be used as an antifungal agent for agricultural applications.

• 생약학회지
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• 제35권1호통권136호
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• pp.62-79
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• 2004

Cell-scattering is a phenotypic change easily observed in most epithelial cells treated with Hepatocyte Growth Factor /scatter Factor (HGF/SF) or phorbol esters (PKC-activators). Recent studies have shown the possibilities to use as therapeutic materials of HGF/SF or non tumor promoting phorbol esters for liver disease, cancer and AIDS. In this study, we tested a cell-scattering activity of 534 methanol extracts from plants inhabiting in Korean peninsula using the phenotype-based assay system. Nine Active extracts were detected : Daphne genkwa, Daphne kiusiana, and Aleurites fordii showed high activity (+++), Euphorbia sieboldiana and Rhodotypos scandens showed medium activity (++), Sambucus sieboldiana var. pendula, Catalpa bignonioides, Sambucus sieboldiana and Lycoris squamigera showed low activity (+). Furthermore, the effects of these active materials in the culture cells were investigated with biochemical studies.

• BMB Reports
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• 제38권6호
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• pp.639-645
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• 2005

Protein kinase C (PKC) isozymes, a family of serine-threonine kinases, are important regulators of cell proliferation and malignant transformation. Phorbol esters, the prototype PKC activators, cause PKC translocation to the plasma membrane in prostate cancer cells, and trigger an apoptotic response. Studies in recent years have determined that each member of the PKC family exerts different effects on apoptotic or survival pathways. $PKC{\delta}$, one of the novel PKCs, is a key player of the apoptotic response via the activation of the p38 MAPK pathway. Studies using RNAi revealed that depletion of $PKC{\delta}$ totally abolishes the apoptotic effect of the phorbol ester PMA. Activation of the classical $PKC{\alpha}$ promotes the dephosphorylation and inactivation of the survival kinase Akt. Studies have assigned a pro-survival role to $PKC{\varepsilon}$, but the function of this PKC isozyme remains controversial. Recently, it has been determined that the PKC apoptotic effect in androgen-dependent prostate cancer cells is mediated by the autocrine secretion of death factors. $PKC{\delta}$ stimulates the release of $TNF{\alpha}$ from the plasma membrane, and blockade of $TNF{\alpha}$ secretion or $TNF{\alpha}$ receptors abrogates the apoptotic response of PMA. Molecular analysis indicates the requirement of the extrinsic apoptotic cascade via the activation of death receptors and caspase-8. Dissecting the pathways downstream of PKC isozymes represents a major challenge to understanding the molecular basis of phorbol ester-induced apoptosis.

• 대한치과교정학회지
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• 제32권3호
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• pp.227-234
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• 2002

본 연구는 조골세포의 특성을 가지고 있는 G292세포주를 이용하여 세포막 이온통로에 대한phorbol ester의 효과를 조사하여 protein kinase C (PCK)의 이온통로에 대한 작용기전을 밝히고자 하였다. Patch clamp 기법을 이용하여 G292 세포에서 cell-attached configuration으로 단일이온통로의 활동을 관찰하고 Phorbol 12, 13-dibutyrate (PDBu)의 효과를 관찰하였다. 안정상태 G292 세포에서 cell-attached 모드로 세포막의 단일이 온통로 활동을 관찰한 결과 45pS의 $K^+$통로가 특징 적으로 우세하였다. 유리 전극 내부에 세포내 액과 세포외 액을 사용하여 전류-전압의 관계를 조사한 결과, 세포내 액을 사용하는 경우에는 역전전압이 5.5mV이었으며 세포외액을 사용하는 경우에는 -27mV이었다. PDBu는 45pS의 이온통로를 10nM이상의 농도에서 이온통로의 열릴 확률을 증가시켰으며 PKC억제제인 staurosporine 10nM에 의하여 차단되는 특성을 보였다. PDBu는 45pS의 이온통로에 작용하여 전류-전압의 관계에서 역전전압을 음의 방향으로 이동시켰으며 동일한 막전압에서 단일이온통로의 전류 크기를 증가시켰다. G292세포에서 PDBu에 의하여 PKC가 활성화되는 것을 western blot으로 확인한 결과 PDBu 0.luM은 세포질에서 세포막으로 PKC translocation을 유의하게 증가시키는 것을 확인하였다. 이상의 결과는 G292세포에서 phorbol ester의 일종인 PDBu가 세포내 PKC를 활성화시켜 45pS의 이온통로를 활성화시키며 이러한 작용의 결과로 세포막전압의 변화가 세포의 기능을 조절할 것으로 사료된다.

• Asian-Australasian Journal of Animal Sciences
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• 제31권6호
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• pp.888-898
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• 2018

Objective: The study was conducted to investigate the effects of replacing soybean meal (SBM) with different levels of detoxified Jatropha curcas kernel meal (DJM) in growing pig diets on growth performance, nutrients digestibility and meat edibility. Methods: A total of 144 pigs with initial body weight of $20.47{\pm}1.44kg$, were randomly allocated to 6 dietary treatments with 6 replications per treatment and 4 pigs per replication for a period of 79 days. Six diets (DJM0, DJM15, DJM30, DJM45, DJM60, and DJM75) were formulated using DJM to replace 0%, 15%, 30%, 45%, 60%, and 75% of SBM. From d 37 to 42, feces and urine were total collected from six barrows in each treatment. At day 79, thirty-six pigs were slaughtered for sampling. The feed intake and weight gain were recorded, while the intestinal morphology, digestive enzyme activities, nutrient digestibility and the content of residual phorbol esters in muscles were determined. Results: The results showed that increasing the replacement of SBM with DJM decreased the parameters including body weight, average daily gain, average daily feed intake, gain-to-feed ratio, weight and villus heights of duodenum, villus height and villus height/crypt depth of jejunum, digestive enzymes (protease, amylase, lipase, and trypsin) activities, and nutrients digestibility (nitrogen deposition, digestibility of nitrogen, energy digestibility, and total nitrogen utilization) (linear, p<0.05; quadratic, p<0.05) and there was no significant difference among DJM0, DJM15, and DJM30 in all measured indices. The highest diarrhea morbidity was observed in DJM75 (p<0.05). Phorbol esters were not detected in pig muscle tissues. Conclusion: The DJM was a good protein source for pigs, and could be used to replace SBM up to 30% (diet phorbol esters concentration at 5.5 mg/kg) in growing pig diets with no detrimental impacts on growth performance, nutrient utilization, and meat edibility.

• 생물정신의학
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• 제2권2호
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• pp.252-256
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• 1995

Corticotropin-releasing factor(CRF)를 분비하는 배양세포주는 CRF의 세포내 조절을 연구하는데 있어 훌륭한 체계가 된다. BE(2)-M17 및 BE(2)-C 세포주는 CRF를 생산하고 분비하며 forskolin 처치에 반응하는 것으로 알려져 있다. 저자들은 이들 세포주에 phorbol 에스테르, 즉 phorbol 12-myristate 13-acetate(TPA) 0.8, 4, 10, 20 nM, 및 nitric oxide(NO)의 신호전달 경로의 자극제(1-${\mu}M$ nitroprusside)와 차단제($1{\mu}M$ nitroprusside+$300{\mu}M$ $N^G$-methyl-D-arginine), 그리고 interleukin-$1{\alpha}$ (IL $1{\alpha}$ ; 4, 20, 100, 500 pM)를 처치하여 CRF를 의미있게 증가시켰으나 세포외 분비는 C 세포에서만 변화되었다. NOS계의 자극제와 차단제는 C 세포의 forskolin 효과를 의미있게 변화시켰다. IL$1{\alpha}$는 두 세포주에 대한 영향이 없었다. 상기 제제들에 대한 이들 세포의 반응이 중추신경계 CRF 신경들과 관련하여 논의되었다.

• Bulletin of the Korean Chemical Society
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• 제30권9호
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• pp.1981-1984
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• 2009

Subcellular localization of protein kinase often plays an important role in determining its activity and specificity. Protein kinase C (PKC), a family of multi-gene protein kinases has long been known to be translocated to the particular cellular compartments in response to DAG or its analog phorbol esters. We used C-terminal green fluorescent protein (GFP) fusion proteins of PKC isoforms to visualize the subcellular distribution of individual PKC isoforms. Intracellular localization of PKC-GFP proteins was monitored by fluorescence microscopy after transient transfection of PKC-GFP expression vectors in the HeLa cells. In unstimulated HeLa cells, all PKC isoforms were found to be distributed throughout the cytoplasm with a few exceptions. PKC$\theta$ was mostly localized to the Golgi, and PKC$\gamma$, PKC$\delta$ and PKC$\eta$ showed cytoplasmic distribution with Golgi localization. DAG analog TPA induced translocation of PKC-GFP to the plasma membrane. PKC$\alpha$, PKC$\eta$ and PKC$\theta$ were also localized to the Golgi in response to TPA. Only PKC$\delta$ was found to be associated with the nuclear membrane after transient TPA treatment. These results suggest that specific PKC isoforms are translocated to different intracellular sites and exhibit distinct biological effects.

• 한국미생물·생명공학회지
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• 제21권1호
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• pp.59-65
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• 1993

방선균 분리주 GMT 1153로부터 ts/NRK 세포의 형태를 변형된 세포형태에서 정상세포의 형태로 전환하는 활성물질을 분리, 정제하였다. 활성물질 MT 1155는 UV, IR, 1H-NMR, 13C-NMR, mass,원소분석 등의 기기분석을 통하여 항진균성 항암물질인 toyocamycin으로 동정되었다. MT 1155는 ts/NRK 세포의 형태전환에 대한 활성외에도 PKA 효소활성 저해와 CTLL 세포에 대한 세포독성이 있으나 K562 소포형성 억제 및 PKC 효소활성에 대한 저해효과도 없었다.

• 한국동물학회지
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• 제36권4호
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• pp.439-451
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• 1993

Protein kinase C isozymes in a human prostate adenocarcinoma PC-3 cell line were characterized. Immunoreactive bands and immunocytochemical stains were obsenred in PC-3 cells with antibodies raised against protein kinase C ${\alpha}$, ${\beta}$, ${\gamma}$, $\delta$, $\varepsilon$, and ζ types, respectively. Protein kinase C ${\alpha}$ corresponded to a immunoreactive band at a molecular weight of 80,000-dalton, whereas molecular weights of other immunoreactive isozvmes of protein kinase C were detected at 68,000-dalton. Protein kinHse C $\delta$ and ζ antibodies detected additional bands at 55,000-dalton and 80,000-dalton, respectively Immunocvtochemical study confirmed the results of the immunoblotting experiments qualitatively: all six protein kinase C isozymes were detected in the cytoplasm of PC-3 cells. Translocation of protein kinase C in PC-3 cells were also examined with phorbol 12-myristate 13-acetate (PMA), bryostatin 2, diolein, and 1-oleoyl-2-acetyl glycerol (OAG). Differential reactions of protein kinase C isozvmes to these activators were obsenred. When PC-3 cells were treated with 10mM bryostatin 2, protein kinase C isozyme u was translocated into the nucleus, whereas s type was translocated into the plasma membrane and the nucleus. Protein kinase C ${\alpha}$ and ζ types were translocated into the nucleus following the treatment with 101M diolein, whereas protein kinase C ${\alpha}$, ${\beta}$, ${\gamma}$, and $\varepsilon$ types were translocated into the nucleus by the treatment with 10mM OAG. Protein kinase C ${\alpha}$ and $\varepsilon$ types were translocated into the nucleus in the presence of 100nM PMA. Protein kinase C $\delta$ type was translocated to the nuclear membrane by these activators, however, only PMA-induced translocation was inhibited by protein kinase C inhibitor, 1-(5-isoquinolinesulfonyll-2-methvlpiperazine dihvdrochloride (H7) . H7 inhibited translocation of protein kinase C ${\alpha}$ type induced by PMA, ${\beta}$ type by OAG and s type by PMA and OAG, whereas it did not affect translocations induced by bryostatin and diolein, respectively. These results suggest that there exist six isoformes of protein kinase C (${\alpha}$, ${\beta}$, ${\gamma}$, $\delta$, $\varepsilon$ and ζ types) in PC-3 cells and that each of these isozvmes distinctivelv reacts to bryostatin, diolein, OAG and PMA, in part due to an altered molecular size and conceivably discrete binding site(s).

• Tuberculosis and Respiratory Diseases
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• 제42권6호
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• pp.862-870
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• 1995

연구배경: 산소기의 작용은 과거에는 세포독성이 주로 알려져 있었던 반면, 최근 들어 산소기의 세포내 신호전달체계에서의 역할에 많은 사람의 관심이 모이고 있다. 여러 cytokine의 전사인자(transcription factor)로 작용하는 $NF{\kappa}B$는 기저상태에서는 세포질에 존재하는데 $I{\kappa}B$와 결합되어 핵내로의 이동이 억제되고 있다. 여러 연구에 의해 $NF{\kappa}B$의 $I{\kappa}B$로부터의 분리는 외부자극에 의해 생성된 산소기에 의한 것으로 알려졌는데, 이렇게 하여 분리된 $NF{\kappa}B$가 핵내로 이동하면 핵내에서 전사인자로 작용하여 여러 유전자의 전사를 증가시키는 것이 보고되었다. IL-8 유전자는 5'flanking promotor region에 $NF{\kappa}B$-like motif가 있어 핵내 $NF{\kappa}B$ activity의 증가로 IL-8 유전자의 전사가 증가되는 것으로 알려졌고, 또한 내독소는 핵내의 $NF{\kappa}B$ activity의 증가와 함께 호중구에서의 산소기의 분비를 가져온다. 이러한 사실로부터 내독소에 의한 IL-8 유전자의 발현은 세포내에서 생성된 산소기에 의해 $NF{\kappa}B$가 $I{\kappa}B$로부터 분리되어 핵내로 이동하고 이로 인해 IL-8 유전자의 전사가 증가되는 가설을 생각할 수 있다. 저자들은 이러한 가설 검정의 첫번째 단계로써 체내 염증반응에서 중요한 역할을 하는 말초혈액 단핵구의 IL-8 및 IL-$1{\beta}$ 유전자 발현에 세포내의 산소기가 관여하는지의 여부를 평가하고자 본 연구를 시행하였다. 방법: Ficoll-Hypaque density gradient 법과 plastic 부착법을 이용하여 말초혈액 단핵구를 분리하였다. 외부에서 투여한 산소기의 농도에 따른 IL-8 및 IL-$1{\beta}$ mRNA 발현의 유무를 관찰하기 위하여 $H_2O_2$를 0, 10, 100, $300{\mu}M/L$, 1mM/L의 농도로 투여하고 6시간이 경과한후 IL-8 및 IL-$1{\beta}$에 대한 Northern blot analysis를 시행하였다. 시간에 따른 IL-8 및 IL-$1{\beta}$ mRNA 변화를 관찰하고자 $H_2O_2$를 $100{\mu}M/L$의 농도로 투여하고 0, 2, 4, 6, 8, 24시간이 경과한 후 Northern blot analysis를 시행하였다. 항산화제가 내독소에 의한 IL-8과 IL-$1{\beta}$ mRNA 발현에 미치는 영향을 평가하기 위하여 TMTU(10 mM/L) 1시간; PDTC($100{\mu}M/L$) 1시간, NAC(10 mM/L) 2.5시간, ME(10mM/(L) 2.5시간, Desferrioxamine(100mM/L) 15시간 동안 전처치 한 디음 내독소를 투여허여 4시간이 경과한 후 IL-8 및 IL-$1{\beta}$ mRNA에 대한 Northern blot analysis를 시행하였다. 결과: $H_2O_2$농도 및 시간에 따른 말초혈액 단핵구에서의 IL-8 및 IL-$1{\beta}$ mRNA의 발현에는 유의한 차이가 관찰되지 않았지만 항산화제로 전처치하였을 때 내독소에 의한 말초혈액 단핵구에서의 IL-8 및 IL-$1{\beta}$ mRNA의 발현이 억제되었고 그 억제정도는 TMTU에서 가장 현저하였다. 결론: 이상의 결과에서 말초혈액 단핵구에서의 IL-8 및 IL-$1{\beta}$ mRNA 발현에 $H_2O_2$가 아닌 다른 산소기가 일부 관여할 것으로 생각된다.