• Title/Summary/Keyword: protein kinase C

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Role of Protein Kinase C in Abnormal Proliferation of Vascular Endothelial Cell induced by 1,2-Dimethylhydrazine; Analysis of Isoform (디메틸히드라진(1,2-Dimethylhydrazine)으로 유도된 혈관내피세포의 비정상적인 증식에서 단백활성효소 시이(Protein Kinase C)의 역할; 동종효소 분석)

  • Lee, Jin;Bae, Yong Chan;Park, Suk Young;Moon, Jae Sul;Nam, Su Bong
    • Archives of Plastic Surgery
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    • v.34 no.1
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    • pp.8-12
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    • 2007
  • Purpose: Protein tyrosine kinase(PTK), protein kinase C(PKC), oxidase, as a mediator, have been known to take a role in signal transduction pathway of angiogenesis. The authors confirmed that PKC is the most noticeable mediator for abnormal proliferation of vascular endothelial cells through in vitro study model using the inhibitors, targeting the formation of three co-enzymes. In this study, we would investigate which isoform of PKC play an important role in abnormal angiogenesis of vascular endothelial cell. Methods: In 96 well plates, $10^4$ HUVECs(human umbilical vein endothelial cells) were evenly distributed. Two groups were established; the control group without administration of DMH(1,2-dimethylhydrazine) and the DMH group with administration of $7.5{\times}10^{-9}M$ DMH. RNA was extracted from vascular endothelial cell of each group and expression of the PKC isoform was analyzed by RT-PCR(reverse transcriptase-polymerase chain reaction) method. Results: RT-PCR analysis showed that $PKC{\alpha}$, $-{\beta}I$, $-{\beta}II$, $-{\eta}$, $-{\mu}$ and $-{\iota}$ were expressed in vascular endothelial cells of each group. DMH incresed the expression of $PKC{\alpha}$ and $PKC{\mu}$, and decreased $PKC{\beta}I$, $PKC{\beta}II$ expression dominantly. Conclusion: Based on the result of this study, it was suggested that $PKC{\alpha}$ and $PKC{\mu}$ may have significant role in abnormal proliferation of vascular endothelial cell.

Expression of protein kinase C in the spinal cords of rats with autoimmune encephalomyelitis (뇌염모델에서 Protein Kinase C의 발현에 관한 연구)

  • Shin, Tae-Kyun;Kim, Hyung-Min;Tanuma, Naoyuki;Matsumoto, Yoh
    • Korean Journal of Veterinary Pathology
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    • v.1 no.1
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    • pp.26-32
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    • 1997
  • Protein kinase C an enzyme of signal transduction has been known to regulate cell proliferation activation as well as apoptosis in some cancer cell lines. To explore the role of PKC in the course of cell mediated autoimmune disease such as experimental autoimmune encephalomyelitis (EAE) EAE was induced in Lewis rats(6-8 weeks old) with immunization of myelin basic protein supplemented with complete Freund's adjuvants and affected spinal cords were sampled at days 13 postimmunization(PI) as peak stage of EAE and at days 21 PI as recovery stage. The spinal cords with EAE were subjected to Northern blot analysis and insitu hybridization of PKC delta which is one of prominant isotypes of PKC in the haematopoietic cells. Northern blot analysis showed that levels of PKS delta mRNA in the spinal cords of rats withEAE was significantly increased at days 13 PI in which inflammatory cells including T cells and macrophages in the EAE lesions appeared. however the stage. By in situ hybridization signals of PKC delta in EAE lesions was intensely expressed on the delta is also expressed on some brain cells in normal rat central nervous system This finding suggests that PKC plays an important role on either activation of inflammatory cells including encephalitogenic T cells and macrophages or apoptotic elimination of some inflammatory cells depending on the stge of EAE.

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The involvement of protein kinase C in the inhibitory effect of methoxamine on the thyrotropin-induced release of thyroxine in mouse thyroid (Mouse 갑상선에서 thyrotropin에 의한 thyroxine 유리에 미치는 methoxamine의 억제효과에 대한 protein kinase C의 관련)

  • Kim, Se-gon;Kim, Jin-sang
    • Korean Journal of Veterinary Research
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    • v.38 no.3
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    • pp.508-517
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    • 1998
  • There is evidence that the sympathetic nervous system exerts a control on thyroid function via an adrenergic innervation of thyroid cells. Although it is clear that the inhibitory effects of catecholamines result from an activation of ${\alpha}_1$-adrenoceptors, the mechanisms involved in ${\alpha}_1$-stimulation are not fully understood. The effects of methoxamine and protein kinase C (PKC) activator on the release of thyroxine ($T_4$) from mouse thyroid were studied to clarify the role of PKC in the regulation of $T_4$ release in vitro. The glands were incubated in the medium, samples of the medium were assayed for $T_4$ by EIA kits. Methoxamine inhibited the TSH-stimulated $T_4$ release. This inhibition was reversed by prazosin, an ${\alpha}_1$-adrenergic antagonist. Futhermore, the inhibitory effect of methoxamine on the $T_4$ release stimulated by TSH was prevented by chloroethylclonidine, an ${\alpha}_{1b}$-adrenoceptor antagonist, but not by WB4101, an ${\alpha}_{1a}$-adrenoceptor antagonist. Also methoxamine inhibited the forskolin-, cAMP- or IBMX-stimulated $T_4$ release. These inhibition were reversed by PKC inhibitors, such as staurosporine and $H_7$. PMA, a PKC activator, completely inhibited the TSH-stimulated $T_4$ release, and its inhibition was reversed by staurosporine and $H_7$, but not by chelerythrine. R59022 (a diacylglycerol kinase inhibitor), like methoxamine, also inhibited the TSH-stimulated $T_4$ release, and its inhibition was also reversed by staurosporine. The present study suggests that methoxamine inhibition of $T_4$ release from mouse thyroid can be induced by activation of the ${\alpha}_{1b}$-adrenoceptors and that it is mediated through the ${\alpha}_1$-adrenoceptor-stimulated PKC formation.

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Detection of the expression of a Bombyx mori Atypical Protein Kinase C in BmPLV-Infected Larval Midgut

  • Cao, Jian;He, Yuanqing;Li, Guohui;Chen, Keping;Kong, Jie;Wang, Fenghua;Shi, Jing;Yao, Qin
    • International Journal of Industrial Entomology
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    • v.22 no.2
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    • pp.59-64
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    • 2011
  • Protein kinase C (PKC) is involved in many cellular signaling pathways, it participates in many physiological processes, such as cell cycle, growth, proliferation, differentiation and apoptosis. To investigate the effect of PKC on the silkworm midgut tissue infection of Bombyx mori parvo-like virus (BmPLV), a B. mori atypical protein kinase C (BmaPKC) gene was cloned from larval midgut tissue, expressed in E. coli and purified. Additionally, the BmPLV susceptible silkworm strain and resistant silkworm strain were used to test the effect of the B. mori infection on BmPLV. The result showed that BmaPKC encodes a predicted 586 amino acid protein, which contains a C-terminal kinase domain and an N-terminal regulatory domain. The maximum expression amount of the soluble (His)6-tagged fusion protein was detected after 0.8 mmol/L IPTG was added and cultured at $21^{\circ}C$. The (His) 6-tagged fusion protein revealed about 73 kDa molecular weight which confirmed by western blot and mass spectrography. Furthermore BmaPKC protein were detected at 0-72 h post-infection in BmPLVinfected larval midgut tissue, western blot showed that as time went on, the expression of BmaPKC increased gradually in susceptible strain, the expression quantity on 72 h is 5 times of 0 h. However, in resistant strain, the expression quantity is slightly lower than susceptible strain. But no significant change in resistant strain was observed as time went on. The available data suggest that BmaPKC may involve in the regulation of BmPLV proliferation.

The Inhibitory Potency of Hesperidin on Protein Kinase C Activity Using a Biochip (바이오칩을 이용한 Protein Kinase C의 활성에 대한 헤스페리딘의 저해 효과)

  • Kang, Jung Ae;Rho, Jong Kook;Choi, Mi Hee;Jung, Young Jin;Park, Sang Hyun
    • Journal of Radiation Industry
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    • v.5 no.1
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    • pp.15-20
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    • 2011
  • Protein kinases are the most important drug targets for the treatment of numerous diseases. The involvement of protein kinase C (PKC) in many biological processes such as development, memory, cell differentiation, and proliferation has been demonstrated. PKC is recognized as an important player in carcinogenesis. Thus, a variety of PKC inhibitors have been investigated. Among them, flavonoids have been demonstrated to affect the activity of many mammalian in vitro enzyme systems. The recent investigation was performed to evaluate the inhibitory effects of hesperidin, which is a flavonoid, on the proliferation and carcinogenesis of many cancers. In this study, an efficient kinase assay based on a biochip using radio-phosphorylation was established and performed for an examination of the inhibitory effects of hesperidin on PKC activity at different concentrations of 50, 200, 500 nM. It was found that hesperidin shows inhibitory potency on PKC, and that the biochip can be used to rapidly screen kinase inhibitors resulting in the therapeutic agents.

The Involvement of Protein kinase C in Glutamate-Mediated Nociceptive Response at the Spinal Cord of Rats (흰쥐의 척수에서 Glutamate가 매개하는 Nociceptive Response에 있어서 Protein kinase C의 관련성)

  • 김성정;박전희;이영욱;양성준;이종은;이병천;손의동;허인회
    • YAKHAK HOEJI
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    • v.43 no.2
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    • pp.263-273
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    • 1999
  • When glutamate was infected intrathecally, the result is similar to those produced by TPA injected. The involvement of protein kinase C (PKC) in the nociceptive responses in rat dorsal horn neurons of lumbar spinal cord was studied. In test with formalin, a PKC inhibitor (chelerythrine) inhibited dose-dependently the formalin-induced behavior response. Neomycin also inhibited it significantly. But, a PKC activator (12-O-tetradecanoylphorbol-13-ester, TPA) showed reverse effect. When gluatamate was injected intrathecally, we observed the result is smilar to those produced by TPA injection. On the other hand, intrathecal injection of glutamate induced thermal and mechanical hyperalgesia. In Tail-flick test, we examined the involvement of PKC on the glutamate-indeced thermal hyperalgesia. Chelerythrine showed an inhibitory effect and TPA enhanced thermal response. Glutamate decreased the mechanical threshold significantly. A pretreatment of chelerythrine and neomycin inhibited glutamate-induced mechanical hyperalgesia, but the effect of neomycin was not significant. TPA had little effect on the mechanical nociceptive response. These results suggest that the PKC activation through metabotropic receptor at postsynaptic region of spinal cord dorsal horn neurons may influence on the persistent nociception produced by chemical stimulation with formalin, thermal and mechanical hyperalgesia induced by glutamate.

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Involvement of Protein Kinase C-δ in Vascular Permeability in Acute Lung Injury

  • Ahn, Jong J.;Jung, Jong P.;Park, Soon E.;Lee, Minhyun;Kwon, Byungsuk;Cho, Hong R.
    • IMMUNE NETWORK
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    • v.15 no.4
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    • pp.206-211
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    • 2015
  • Pulmonary edema is a major cause of mortality due to acute lung injury (ALI). The involvement of protein kinase C-${\delta}$ (PKC-${\delta}$) in ALI has been a controversial topic. Here we investigated PKC-${\delta}$ function in ALI using PKC-${\delta}$ knockout (KO) mice and PKC inhibitors. Our results indicated that although the ability to produce proinflammatory mediators in response to LPS injury in PKC-${\delta}$ KO mice was similar to that of control mice, they showed enhanced recruitment of neutrophils to the lung and more severe pulmonary edema. PKC-${\delta}$ inhibition promoted barrier dysfunction in an endothelial cell layer in vitro, and administration of a PKC-${\delta}$-specific inhibitor significantly increased steady state vascular permeability. A neutrophil transmigration assay indicated that the PKC-${\delta}$ inhibition increased neutrophil transmigration through an endothelial monolayer. This suggests that PKC-${\delta}$ inhibition induces structural changes in endothelial cells, allowing extravasation of proteins and neutrophils.

Effects of muscarinic receptor stimulation on the thyrotropin-induced thyroxine release in the guinea pig thyroid (기니픽 갑상선에서 Thyrotropin에 의한 thyroxine 유리에 대한 muscarinic 수용체 자극효과)

  • Kim, Hong-hyun;Kim, Jin-shang
    • Korean Journal of Veterinary Research
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    • v.39 no.1
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    • pp.55-61
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    • 1999
  • The present experiments were performed to examine the effects of acetylcholine (ACh) and carbachol (CC) on thyroxine ($T_4$) release and any possible relation between inhibition of $T_4$ release and signaling pathway in guinea pig thyroids. The thyroids were incubated in the medium containing the test agents, samples of the medium were assayed for $T_4$ by EIA kits. ACh and CC inhibited the TSH-stimulated $T_4$ release. These inhibition were reversed by atropine, but not by d-tubocurarine. The inhibitory effects of ACh on $T_4$ release were prevented by $M_{1^-}$ and $M_{3^-}$muscarinic antagonists and its inhibition was also slightly reversed by $M_{2^-}$ and $M_{4^-}$muscarinic antagonists. R59022, like ACh and CC, also inhibited the TSH-stimulated $T_4$ release. This inhibition was reversed by protein kinase C inhibitor and $Ca^{2+}$ channel blocker. The present study suggests that cholinergic inhibition of $T_4$ release from thyroids can be induced mainly by activation of the $M_{1^-}$ or $M_{3^-}$ receptors and that it is mediated through the muscarinic receptorstimulated protein kinase C activation.

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Action of Phospholipase $A_2$in Histamine Release from Mast Cells (비만세포에서 Histamine유리에 관여하는 Phospholipase $A_2$의 작용)

  • 이윤혜;이승준;서무현;장용운;윤정이
    • YAKHAK HOEJI
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    • v.45 no.3
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    • pp.287-292
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    • 2001
  • To investigate whether phospholipase $A_2$pathway is involved in histamine release of rat peritoneal mast cells, we measured histamine release in the presence of various enzyme inhibitors involved in eicosanoid pathway, such as phospholipase $A_2$, cyclooxygenase and lipoxygenase. Phospholipase $A_2$inhibitors, manoalide and OPC, significantly inhibited histamine release induced by 100 $\mu$M ATP and 1$\mu$g/ml compound 48/80. Cyclooxygenase inhibitors, ibuprofen and indomethacin, significantly inhibited ATP-induced histamine release and lipoxygenase inhibitors, baicalein and caffeic acid, also significantly inhibited. To investigate the involvement of protein kinase in ATP- and compound 48/80-induced histamine release, we observed effects of protein kinase inhibitors on histamine release. Bisindolmaleimide (protein kinase C antagonist) dose-dependently inhibited both ATP and compound 48/80-induced histamine release. Tyrosine kinase inhibitors (methyl 2,5-dihydroxy cinnamate and genistein) dose-dependently inhibited ATP and compound 48/80-induced histamine release. Protein kinase C and tyrosine kinase seem to be involved in histamine release induced by ATP and compound 48/80. These results suggest that phospholipase $A_2$pathway as well as protein kinase C and tyrosine kinase are involved in histamine release of rat peritoneal mast cells by ATP and compound 48/80.

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Isoforms of Protei,n Kinase C during the Differentiation of Chick Limb Mesenchvme (계배 간충직세포 분화과정에서의 Protein Kinase C Isoform들의 변화)

  • 손종경;강신성
    • The Korean Journal of Zoology
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    • v.38 no.2
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    • pp.286-293
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    • 1995
  • The present studies were undertaken to examine the activitites of PKC isoforms in cultures of chick limb mesenchvme. Micromass cultures were prepared using wing buds of stage 23/24 (Hamburger and Hamilton, 19511 chick embryo. The cells were homogenized and DEAE-cellulose column chromatography was performed to get fraction containing protein kinase C (PKC) activity. PKC isoforms were resolved with hvdroxyapatitie column chromatography. Profile of PKC isoforms of cultures were compared with that of rat brain. Activity of $PKC-\beta$ isoform was appeared at the early stage of chondrogenesis. On 3 daw of culture, activities of both PKC a and $\beta$ were observed with remarkable increase but no activity of y isoform was appeared. Treatment of phorbol-12-mvristate-13-acetate (PMA) (10-7 M) to the culture inhibited chondrosenesis and down-regulated a and $\beta$ isoforms. Staurosporine promoted chondro!genesis without any effect on PKC isioforms profile. These data indicate that PKC a and $\beta,$ especiallv $\beta$ isoform is related to chondrosenesis and the promoting effect of staurosporine on chondrogenesis is not related to PKC isoforms activities.

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