• Bulletin of the Korean Chemical Society
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• 제29권10호
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• pp.2023-2025
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• 2008

To evaluate the potential of radioiodine labelled hypericin as a malignant glioma imaging agent, U-251 MG, U-373 MG, C6 glioma and fibroblast were treated with a I-123 labelled hypericin derivative and C6 glioma transplanted nude mouse were injected with a I-124 labelled hypericin derivative for a micro PET imaging. 2- Iodohypericin was prepared as a reference compound. In this paper, we describe the syntheses of 2- iodohypericin and 2-[$^{123}I/^{124}I$]iodohypericin and the results of a corresponding biological evaluation. In all glioma cell lines, 2-[$^{123}I$]iodohypericin uptake was increased in a time dependant manner and an accumulation of 2-[$^{124}I$]iodohypericin was observed in C6 glioma bearing nude mouse. These results suggest that radioiodine labelled hypericin can visualize a PKC overexpressed malignant glioma.

• BMB Reports
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• 제47권10호
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• pp.552-557
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• 2014

The main purpose of this study was to investigate whether type 3 muscarinic acetylcholine receptor (M3R) dysfunction induced vascular hyperpermeability. Transwell system analysis showed that M3R inhibition by selective antagonist 4-diphenylacetoxy-N-methylpiperidine methiodide (4-DAMP) and small interfering RNA both increased endothelial permeability. Using coimmunoprecipitation and Western blot assay, we found that M3R inhibition increased VE-cadherin and ${\beta}$-catenin tyrosine phosphorylation without affecting their expression. Using PTP1B siRNA, we found that PTP1B was required for maintaining VE-cadherin and ${\beta}$-catenin protein dephosphorylation. In addition, 4-DAMP suppressed PTP1B activity by reducing cyclic adenosine monophosphate (cAMP), but not protein kinase $C{\alpha}$ ($PKC{\alpha}$). These data indicate that M3R preserves the endothelial barrier function through a mechanism potentially maintaining PTP1B activity, keeping the adherens junction proteins (AJPs) dephosphorylation.

• 대한한방부인과학회지
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• 제21권2호
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• pp.59-75
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• 2008

Purpose: This study was performed to determine the inhibitory effect of Polygonum multiflorum(PM) on melanin synthesis in B16F10. Methods: The Inhibitory effects of Polygonum multiflorum(PM) on melanin synthesis were determined by in-vitro assay. To elucidate inhibitory effects of Polygonum multiflorum on melanin synthesis, we determined the melanin release and melanin production in B16F10. And to investigate the action mechanism, we assessed the gene expression of tyrosinase, TRP-1, TRP-2, MMP-2, PKA, PKC, ERK-1 ERK-2, AKT-1, MITF in B16F10. Results: 1. PM inhibited melanin-release, melanin production in B16F10. 2. PM inhibited tyrosinase activity in vitro and in B16F10. 3. PM suppressed the expression of tyrosinase, TRP-1 in B16F10. 4. PM suppressed the expression of PKA in B16F10. 5. PM suppressed the expression of ERK-1, ERK-2, AKT-1 in B16F10. 6. PM suppressed the expression of MITF in B16F10. Conclusion: From these results, it may be concluded that PM possesses the antimelanogenetic effects.

• 동의생리병리학회지
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• 제17권5호
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• pp.1335-1338
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• 2003

We have previously reported that the vasodilatory effect of BanhabackchuIChunma-tang(半夏白朮天麻湯; BCT), a herbal formula, and its mechanism might be associated with at least in part NO pathway. In the present study, we studied the influence of BanhabackchulChenuma-tang (BCT) on the phosphorylation of LC20, in parallel, the distribution of α-protein kinase C(PKCα) by phenylephrine was monitored using laser scan confocal immunofluorescent microscopy in freshly isolated ferret portal vein smooth muscle living single cells. Phenylephrine stimulation induced LC20 phosphorylation and translocation of PKCα. However, BCT dephosphorylated LC20 phosphorylation and inhibited the translocation of PKCα. Our results demonstrate that the mechanism of relaxant effect of BanhabackchulChunma-tang inhibition is associated with inhibition of PKCα activation and LC20 phosphorylation.

• Bioinformatics and Biosystems
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• 제2권1호
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• pp.28-32
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• 2007

The protein phosphorylation is one of the important processes in the cell signaling pathway. A variety of protein kinase families are involved in this process, and each kinase family phosphorylates different kinds of substrate proteins. Many methods to predict the kinase-specific phosphoryrated sites or different types of phosphorylated residues (Serine/Threonine or Tyrosin) have been developed. We employed Supprot Vector Machine (SVM) to attempt the prediction of protein kinase specific phosphorylation sites. 10 different kinds of protein kinase families (PKA, PKC, CK2, CDK, CaM-KII, PKB, MAPK, EGFR) were considered in this study. We defined 9 residues around a phosphorylated residue as a deterministic instance from which protein kinases determine whether they act on. The subsets of PSI-BALST profile was converted to the numerical vectors to represent positive or negative instances. When SVM training, We took advantage of multiple SVMs because of the unbalanced training sets. Representative negative instances were drawn multiple times, and generated new traing sets with the same positive instances in the original traing set. When testing, the final decisions were made by the votes of those multiple SVMs. Generally, RBF kernel was used for the SVMs, and several parameters such as gamma and cost factor were tested. Our approach achieved more than 90% specificity throughout the protein kinase families, while the sensitivities recorded 60% on average.

• 대한약리학회지
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• 제32권2호
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• pp.211-219
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• 1996

The inhibitory effect of cyclosporin A (CsA) on nitric oxide production is not related to the immunosuppressive action of the drug, but to the renal toxicity and arterial hyper-tension. In this study the experimental interventions to reverse the inhibition of nitric oxide production by cyclosporin A in rat aortic smooth muscle cells were examined. CsA inhibited the accumulation of nitrite, the stable end product of nitric oxide, in culture media in a concentration $(0.1{\sim}100{\mu}g/ml)-dependent$ manner. The inhibitory effect of CsA on nitrite accumulation were not antagonized by arginine (10 mM), a substrate of nitric oxide synthase, nor by calcium ionophore A23187 $(7{\mu}M)$. Forskolin, an activator of adenylate cyclase, which enhanced iNOS induction at transcriptional level, completely reversed the inhibitory action of CsA on nitrite accumulation. However, PMA (2 nM) and PDB (50 nM), PKC activators, increased the inhibitory action of CsA on nitrite accumulalion. From these results, it is suggested that cyclic AMP-elevating agents may be candidates of therapeutic agents in prevention and treatment of renal toxicity and arterial hypertension induced by CsA. Among conventional antihypertensive drugs, calcium channel blockers and ${\alpha}-blockers$ are preferred to ${\beta}-blockers$.

• 한방안이비인후피부과학회지
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• 제16권3호
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• pp.96-115
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• 2003

Introduction and Objectives : Administrating the Sopungchengyoungtanggami-bang (SCG) extract on to the atopic dermatitis(AD) is to study change of external dermal formation, change of leukocytes in vasculature, change of lipid formation in stratum corneum and distribution of ceramide and this study is done through forcing injury to rat's back skin which are lipid protect formation in stratum corneum. Materials and Methods : The AD which caused intentionally using the external application on the rat's back skin was used the SCG. The change of leukocytes in vasculature has been identified through optima 5.2 and student t-test and the results were made into dermal formation graph. Results : After dispensing SCG extract into the AD, the dermal injury was decreased. Especially, recover of lipid protection formation which include lipid and ceramide in stratum corneum is suppressing acute inflammation that some factors are PKC, TNF-${\alpha}$, IL-12B which controled the secretion of relating inflammatory cytokine, also went onto decrease of angiogenesis, and the decrease of degranulated mast cell. In addition, the decrease of epithelial injury also caused the growth of cell to decrease in stratum basale and cytoclasis. In the vasculature, the leukocytes were also decreased and it could relate to decrease AD Conclusions : Thus, SCG has effect on AD suppressing the dermal injury through recovering of lipid protection formation in stratum corneum.

• Archives of Pharmacal Research
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• 제18권4호
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• pp.262-266
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• 1995

Bacterial lipopolysaccharide (LPS) is one of the most potent inducers of various cytokines nad other proinflammatory mediators in macrophages. Although pathophysiological consequences of LPS-induced responses are well established, the mechanisms through which LPS-generated singals are transduced remain unclear. In the present study, we attempted to determine early intracellular events after LPS binding which transduced the signal for the induction of arachidonic acid metabolism in rat alveolar macrophages. While H-7, a protein kinase C(PKC) inhibitor, did not affect LPS-stimulated prostaglandin synthesis, staurosporine enhanced archidonic acid etabolism in macropahages treated with LPS. Phorbol-12-myristate-13 acetate snesitive to LPS compare with control group. PMA and H-7 did not alter the effect of flucose. Pertussis toxin did not show nay effect, thus pertussis toxin snesitive G-protein pathway appears not to play a role in this experimental system. Genistein and tyrphostin 25, protein tyrosine kinase 9PTK) inhibitors, markedly inhibited prostaglandin synthesis in macrophages nal transduction events leading to icnreased macrophage arachidonic acid metabolism.

• 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.

• 한국생물물리학회:학술대회논문집
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• 한국생물물리학회 2003년도 정기총회 및 학술발표회
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• pp.53-53
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• 2003

Calumenin was previously identified as a high affinity Ca$\^$2+/ binding protein in mouse cardiac sarcoplasmic reticulum (SR). For the present study, a 48 kDa skeletal homologue of calumenin was identified by sucrose-density gradient of rabbit skeletal SR membranes, concanavalin A treatment, 2D-gel electrophoresis, $\^$45/Ca$\^$2+/ overlay, Stains-all staining, and MALDI-TOF analysis. We attempted to clone the skeletal calumenin by RT-PCR based on mouse cardiac and human calumenin sequences. The deduced amino acid sequence (315 residues) of the skeletal calumenin showed high identity to mouse cardiac calumenin (90％). As seen in the cardiac calumenin, the deduced sequence contains a 19 amino acid N-terminal signal sequence and a HDEF C-terminal sequence, a putative retrieval signal to ER. Also, the skeletal calumenin contains one N-glycosylation site, three PKC phosphorylation sites, eight casein kinase 2 phosphorylation sites, and 6 EF-hand domains. GST-calumenin showed a conformational change and increased mobility in the presence of Ca$\^$2+/ in SDS-PAGE. Three calumenin interacting proteins (ryanodine receptor 1, glycogen phosphorylase, and phosphofructo kinase) were identified by pull-down assay with GST-calumenin and solubilized SR. All the interactions were Ca$\^$2+/dependent. The present results suggest that calumenin plays an important role in Ca$\^$2+/ homeostasis of muscle cells.