• 제목/요약/키워드: protein kinase C

검색결과 453건 처리시간 0.116초

카바마제핀 장기 투여가 백서(白鼠) 뇌의 Protein Kinase A와 Protein Kinase C 활성도에 미치는 영향 (The Effects of Chronic Carbamazepine Administration on Protein Kinase A and Protein Kinase C Activities in Rat Brain)

  • 임두원;김인;서광윤
    • 생물정신의학
    • /
    • 제5권2호
    • /
    • pp.227-234
    • /
    • 1998
  • Objective : Many evidences suggest that patients with bipolar disorder have functional abnormalities in their postreceptor signal transduction pathways, and mood stabilizing effect of lithium is exerted by modulating this dysfunctioning system. Carbamazepine, an antiepileptic agent, is also known to be effective in the treatment and prevention of bipolar disorder. But the precise mechanism of action of the drug is still poorly understood. This study was performed to elucidate the possible therapeutic mechanism of carbamazepine. Method : The effects of chronic carbamazepine administration on protein kinase A and protein kinase C activities in frontal cortex of rat brain after 2 weeks of drug administration were measured and compared with those of control subjects. Results : Mean(${\pm}SE$) value of activity(phosphate transfer ${\mu}mol/mg$ of $protein{\cdot}min$) of protein kinase A in control and test group was $0.249563{\pm}0.036$ and $0.539853{\pm}0.078$, and that of protein kinase C was $0.654817{\pm}0.053$ and $1.146205{\pm}0.052$ respectively, being increased in test group. And differences between the two groups were statistically significant for both enzymes(protein kinase A ; p<0.01, protein kinase C ; p<0.001). Conclusion : These results show that chronic carbamazepine administration increases protein kinase A and C activities, and concerning the possible mode of therapeutic action in bipolar disorder it is suggested that enhanced enzymes phosphorylate receptor-G-protein-effector complexes to dampen hyperfunctioning neuronal activity and thus stabilize the system.

  • PDF

말초혈액 단핵구에 대한 내독소 자극의 신호 전달에서 Protein Kinase C와 Protein Tyrosine Kinase의 역할 (The Role of Protein Kinase C and Protein Tyrosine Kinase in the Signal Transduction Pathway of Stimulus Induced by Endotoxin in Peripheral Blood Monocyte)

  • 김재열;박재석;이귀래;유철규;김영환;한성구;심영수
    • Tuberculosis and Respiratory Diseases
    • /
    • 제44권2호
    • /
    • pp.338-348
    • /
    • 1997
  • Background : Endotoxin, the component of outermembrane of gram negative organism, plays an important role in the initiation and amplification of inflammatory reaction by its effects on inflammatory cells. Until recently, there have been continuing efforts to delinate the mechanisms of the signal trasduction pathway of endotoxin stimuli on inflammatory cells. By uncovering the mechanisms of signal transduction pathway of endotoxin stimuli, we can expect to have tools to control the excessive inflammatory responses which sometimes may be fatal to the involved host. It was generally accepted that endotoxin exerts its inflammatory effects through inflammatory cytokines that are produced by endotoxin-stimulated inflammatory cells and there were some reports on the importance of protein kinase C and protein tyrosine kinase activation in the production of inflammatory cytokines by endotoxin So we evaluated the effect of pretreatment of protein kinase C inhibitors (H7, Staurosporin) and protein tyrosine kinase inhibitors(Herbimycin, Genistein) on the endotoxin-stimulated cytokines(IL-8 & TNF-$\alpha$) mRNA expression. Method : Peripheral blood monocytes were isolated from healthy volunteers by Ficoll-Hypaque density gradient method and purified by adhesion to 60mm Petri dishes. Endotoxin(LPS 100ng/ml) was added to each dishes except one control dish, and each endotoxin-stimulated dishes was preincubated with H7, Staurosporin(protein kinase C inhibitor), Herbimycin or Genistein(protein tyrosine kinase inhibitor) respectively except one dish. Four hours later the endotoxin stimulation, total RNA was extracted and Northern blot analysis for IL-8 mRNA and TNF-$\alpha$ mRNA was done. Result : Endotoxin stimulation increased the expression of IL-8 mRNA and TNF-$\alpha$ mRNA expression in human peripheral blood monocyte as expected and the stimulatory effect of endotoxin on TNF-$\alpha$ mRNA expression was inhibited by protein kinase C inhibitors(H7, Staurosporin) and protein tyrosine kinase inhibitors (Herbimycin, Genistein). The inhibitory effect of each drugs was increased with increasing concentration. The stimulatory effect of endotoxin on IL-8 mRNA was also inhibited by H7 and protein tyrosine kinase inhibitors (Herbimycin, Genistein) dose-dependently but not by Staurosporin. Conclusion : Protein kinase C and protein tyrosine kinase are involved in the endotoxin induced signal transduction pathway in human peripheral blood monocyte.

  • PDF

Protein Kinase 억제제 첨가 후 Platelet-Activating Factor에 의하여 자극된 호중구반응의 변경 (Alteration of the Activated Responses in Platelet-Activating Factor-Stimulated Neutrophils by Protein Kinase Inhibitors)

  • 이강건;고지영;함동석;신용규;이정수
    • 대한약리학회지
    • /
    • 제32권1호
    • /
    • pp.103-112
    • /
    • 1996
  • Platelet-activating factor (PAF)에 의하여 자극된 호중구 respiratory burst, 탈과립과 세포질 칼슘농도의 증가에 있어 protein kinase C와 protein tyrosine kinase의 역할을 관찰하였다. PAF에 의하여 자극된 호중구에서 superoxide 및 $H_2O_2$의 생성과 myeloperoxidase와 acid phosphatase의 유리는 protein kinase C 억제제인 staurosporine과 H-7 그리고 protein tyrosine kinase 억제제인 genistein과 tyrphostin에 의하여 억제되었다. PAF에 의한 호중구 세포내 칼슘농도의 증가는 staurosporine, genistein과 methyl-2,5-dihydroxycinnamate에 의하여 억제 되었다. Staurosporine은 PAF에 의하여 자극된 호중구에서 세포내 칼슘유리와 망간유입을 억제 하였다. Genistein과 methyl-2,5-dihydroxycinnamate는 PAF에 의한 망간유입을 억제하였으나, 세포내 칼슘유리에 대한 이들의 효과는 관찰되지 않았다. PMA에 의하여 활성화된 호중구에서 세포내 칼슘농도의 증가에 대한 PAF의 자극효과는 감소되었다. Protein kinase C와 protein tyrosine kinase는 PAF에 의하여 자극된 호중구에서의 respiratory burst, lysosomal enzyme유리와 칼슘동원에 관여할 것으로 제시된다. 세포내 칼슘농도의 증가는 protein kinase의 영향을 다르게 받는 세포내 칼슘유리와 세포외부로 부터의 칼슘유입에 의하여 이루어질 것으로 추정된다. Protein kinase C가 활성화되어 있는 상태에서 세포내 칼슘동원에 대한 PAF의 자극작용은 감소될 것으로 시사된다.

  • PDF

Potential Role of Protein Kinase C on the Differentiation of Erythroid Progenitor Cells

  • Lee, Sang-Jun;Cho, In-Koo;Huh, In-Hoe;Yoon, Ki-Yom;Ann, Hyung-Soo
    • Archives of Pharmacal Research
    • /
    • 제18권2호
    • /
    • pp.90-99
    • /
    • 1995
  • The effect of protein kinase C inhibitors, sturosporine and 1-(5-isoquinolinyl sulfonyl)-2-methyl piperazine(H7) on in vitro differentiation of erythroid progenitor cells which were isolated from spleens of mice infected with the anemia-inducing strain of Friend virus were examined. Erythropoietin-mediated differentitation of erythroid progenitor cells, as determined by the incorporation of $^{59}Fe$ into protoporphyrin, was inhibited by staurosporine and H7 in a concentration -dependent manner. Scatchard analysis of the $^3H-phorbol-12$, 13-dibutyrate binding to erythroid progenitor cells revealed that at the high affinity sites the dissociation constant was 22nM and the maximum number of $^3H-phorbol-12$, 13-dibutyrate binding to erythroid progenitor cells revealed that at the high affinity sites the dissociation constant was 22nM and the maximum number of $^3H-phorbol-12$, 13-dibutyrate binding sites per cell was approximately $3.7\times10^5$. Cytosonic protein kinase C was isolated from erthroid progenitor cells and then purified by sequential column chromatogrphy. Two isoforms of protein kinase C were found. Photoaffinity labeling of the purified protein kinase C samples with $^3H-phorbol-12$12-myristate 13-acetate followed by analysis of SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and autofluorography showed radiolabeled 82-KDa pepticles. Rediolabeling of the 82-KDa peptides with $^3H-phorbol-12$myristate 13-acete was almost completely blocked by excess unlabeled phorbol 12-myristate 13-acetate was almost 12-muristate 13-acetate-promoted phosphorylation with the puyrified protein kinase C samples showed that the phosphorylation of 82-KDa peptides was increased as the concentration of phorbol 12-myristate 13-acetate was increased from $10^{-8}M{\;}to{\;}10^{-4}$M. In light of the findings that erythroid progenitor cells possessed an abundance of protein kinase C and that stauroporine and H7 inhibited erythroid differentiation, it seemed likely that protein kinase C would play a role in the erythroid progenitor cell development.

  • PDF

G292 세포에서 $K^+$통로에 대한 phorbol ester의 효과 (Effect of Phorbol ester on $K^+$channel in an G292 osteoblast-like cell)

  • 김미경;박수병
    • 대한치과교정학회지
    • /
    • 제32권3호
    • /
    • pp.227-234
    • /
    • 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.

백서의 심방관류모델에서 protein kinase C의 활성화가 atrial natriuretic peptide(ANP) 조절에 미치는 영향 (Effects of activation of protein kinase C on the regulation of atrial natriuretic peptide(ANP) by isolated perfused left atria)

  • 강창원;강형섭;이호일
    • 대한수의학회지
    • /
    • 제37권4호
    • /
    • pp.735-744
    • /
    • 1997
  • 심방근 세포는 심방이뇨호르몬을 합성, 저장 그리고 분비하며, 세포내외 이온의 농도, 수분균형 및 혈압 등을 조절하는 것으로 알려져 있다. 또한 심방근의 인장자극에는 Atrial Natriuretic Peptide(ANP)를 2단계(분비, 유리)의 과정으로 이루어져 있으며, 이에 따른 심방이뇨호르몬의 분비 조절기전에 대하여서는 명확히 알려져 있지 않다. 따라서 본 연구는 백서의 심방근 적출관류 모델을 이용하여 protein kinase C와 ANP 조절의 상관관계를 밝히고 분비와 유리의 과정중 어떠한 과정을 이용하여 분비자극에 영향을 주는지를 관찰하기 위하여 본 실험을 실시하였다. PKC 활성제인 PMA(phorbol 12-mystrate 13-acetate)는 ANP의 유리를 현저하게 증가시켰으며, PKC 억제제인 H-7(1-(5-isoquinoline sulfonyl)-2-methyl piperazine dihydrochlo-ride)에 의해 유리를 억제시켰다. PMA와 H-7을 동시에 처리한 경우 PMA에 의하여 증가된 ANP의 유리가 H-7에 의하여 차단됨을 관찰할 수 있었다. 따라서 백서의 관류 심방에서의 ANP 분비 증가는 PKC 활성화에 의하여 이루어지며, ANP분비의 2단계중 ANP 유리에 영향을 줌을 알 수 있었다.

  • PDF

허혈이 유도된 대뇌신경세포에 대한 항산화제 및 Ampa/kainate 수용체 길항제의 영향 (Effect of Antioxidant and Ampa/kainate Receptor Antagonist on Cerebral Neurons Damaged by Ischemia)

  • 오연균
    • 동의생리병리학회지
    • /
    • 제19권4호
    • /
    • pp.1022-1026
    • /
    • 2005
  • To clarify the toxic effect on cultured neonatal mouse cerebral neurons damaged by ischemia, we examined the cytotoxicity induced by ischemia and the protective effect of antioxidant and AMPA/kainate receptor antagonist against ischemia-induced cytotoxicity on cultured cerebral neurons. For this study, mice were administrated with 20ug/kg cyclothiazide or 50U/kg vitamin E via intraperitoneal injection for 2 hours before ischemic induction. After cell culture for 7 days, cell viability, amount of neurofilament and protein kinase C activity were examined. Ischemia decreased significantly cell viability, amount of neurofilament and the increase of protein kinase C activity in these cultures. In the protective effect, vitamin I showed remarkably the increase of cell viability and amount of neurofilament, and the decrease of protein kinase C activity but, cyclothiazide did not showed any protective effect on ischemia-induced cytotoxicity. From these results, it is suggested that vitamin I is effective in blocking the neurotoxicity induced by ischemia, but cyclothiazide as a AMPA/kainate receptor antagonist is not.

근세포 분화에 관한 연구 계배의 Myoblasts에 있어서 Protein Kinase C (PKC)의 인 산화작용과 Down Regulation (Studies on the Differentiation of Skeletal Muscle Cells in uitro : The Phosphorylation and Down Regulation of Protein Kinase C in Myoblasts of Chick Embryos)

  • 문현근;최원철
    • 한국동물학회지
    • /
    • 제35권2호
    • /
    • pp.161-172
    • /
    • 1992
  • In the short-term treahent of 12-0-tetradecanoylphorbol-13-acetate (TPA) or platelet-derived growth factor (PDGF), the'Wh and PDGF induced the Protein Kinase C (PKC) activation and migration from the cytoplasm to the peripheral nulcear membrane. And the activated PKC which was directly or indirectly stimulated by TPA or PDGF Phosphorylated many kinds of PKC's targeting proteins and induces various biological responses. Especially, the cytoplasmic PKC was phosphorylated within 1 hr and 10 min by TPA-and PDGF-treahent respectivelv. In the long-term treatment of TPA or PDGF, both of them induced the down-regulation and translocation of PKC in the mvoblasts. The down-regulation of PKC isozyrnes, the pattern of PKC I and ll was similar to the PKC 111 isozpnes in the cytoplasm. But in the nucleolus, the TPA did not induce and down-regulation or the inhibition of the immunoreactivity of PKC III antibody. This investigation indicates that each isozvmes of PKC mal be performed the different effects to the down-regulation of the cytoplasm or nucleolus. And douvn-regulated myoblasts contained low immunoreactivity of PKC antibodies.

  • PDF

말 정소내 protein kinase C의 발현 (Expression of protein kinase C in the testes of horse)

  • 진재광;신태균
    • 대한수의학회지
    • /
    • 제38권1호
    • /
    • pp.9-15
    • /
    • 1998
  • To investigate the involvement of protein kinase C(PKC) isoenzyme in the testes which control spermatogenesis and hormone secretion, we examined cellular distribution of four types of PKC $\alpha$, ${\beta}I$, ${\delta}$ and ${\theta}$ in the horse testes using PKC antisera by western blot analysis and immunohistochemistry. By the western blot analysis, PKC $\alpha$ and ${\beta}I$ were detected at 82KD, while PKC ${\delta}$ and ${\theta}$ were detected at 80KD in the testes of both juvenile and adult horses. In juvenile horse, PKC $\alpha$, ${\delta}$ and ${\theta}$ except ${\beta}I$ were not detected in the cells of the testes, whereas PKC ${\beta}I$ was immunoreacted with only in spermatocytes. In adult, PKC $\alpha$, ${\beta}I$, ${\delta}$ and ${\theta}$isoenzymes were localized in interstitial cells of the testes. In the seminiferous tubules, PKC ${\beta}I$ is localized in spermatocyte, spermatid and spermatozoa, while PKC ${\delta}$ is localized only in spermatids. We suggest that this is a first report to localize PKC in the testes of horse and PKC isoenzymes are upregulated in the cells of horse testes depending on ages. These findings also suggest that certain PKC isoenzyme plays an important role in the signal transduction of spermatogenic cells and interstitial cells in horse testes.

  • PDF