• Archives of Pharmacal Research
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• 제23권3호
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• pp.202-205
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• 2000

A convenient pathway for synthesis of trans-metanicotine analogues was developed. trans-Metanicotine, a subtype(${\alpha}4{\beta}2$)-selective ligand for neuronal nicotinic acetylcholine receptor, is under clinical phase for Alzheimer's disease. Zn-mediated allylation of allyl bromide and acetaldehyde followed by Heck reaction with 3-bromopyridine gave 5-pyridin-3-yl-pent-4-en-3-ol (2). Tosylation of 5-pyridin-3-yl-pent-4-en-3-ol followed by substitution reaction with methylamine in sealed tube gave methyl-(1-methyl-4-pyridin-3-yl-but-3-enyl)-amine (4) in good yields. Thus, trans-metanicotine analogues modified at the $\alpha$-position of the methylamino group with various functional groups can be obtained in 4 steps.

• BMB Reports
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• 제42권8호
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• pp.516-522
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• 2009

SH2D4A, comprising a single SH2 domain, is a novel protein of the SH2 signaling protein family. We have previously demonstrated SH2D4A is expressed ubiquitously in various tissues and is located in the cytoplasm. In this study we investigated the function of SH2D4A in human embryonic kidney (HEK) 293 cells using interaction analysis, cell proliferation assays, and kinase activity detection. SH2D4A was found to directly bind to estrogen receptor $\alpha$ (ER$\alpha$), and prevent the recruitment of phospholipase C-$\gamma$ (PLC-$\gamma$) to ER$\alpha$. Moreover, we observed its inhibitory effects on estrogen-induced cell proliferation, involving the protein kinase C (PKC) signaling pathway. Together, these findings suggested that SH2D4A inhibited cell proliferation by suppression of the ER$\alpha$/PLC-$\gamma$/PKC signaling pathway. SH2D4A may be useful for the development of a new anti-cancer drug acting as an ER signaling modulator.

• 대한의생명과학회지
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• 제13권2호
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• pp.119-125
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• 2007

It is widely known that protein tyrosine kinases (PTKs) are involved in controlling many biological processes such as cell growth, differentiation, proliferation, survival and apoptosis. An $\alpha3\beta4$ subunit combination acts as a major functional acetylcholine receptor (nAChRs) in male rat major pelvic ganglion (MPG) neurons, and their activation induces fast inward currents and intracellular calcium increases. Recently it has been reported that the activity of acetylcholine receptors (AChRs) in some neurons can be negatively regulated by PTKs. However, the exact mechanism of regulation of nAChRs by PTKs is poorly understood. Therefore, we examined the potential role particular in nAChR by PTK using electrophysiology and calcium imaging in male rat MPG neurons. ACh induced inward currents and $(Ca^{2+})_i$ increases in MPG neurons, concomitantly. These responses were inhibited by more than 90% in $Na^+$- or $Ca^{2+}$- free solution. $\alpha$-conotoxin AuIB, a selective $\alpha3\beta4$ nAChR blocket, inhibited ACh-induced inward currents. Genistein (10 $\mu$M), a broad-spectrum tyrosine kinase inhibitor, markedly decreased ACh-induced currents and $Ca^{2+}$ transients, whereas 10 $\mu$M genistin, an inactive analogue, had little effect. Overall these data suggest that the activities of $\alpha3\beta4$ AChRs in MPG neurons are positively regulated by PTK. In conclusion, trosine kinase may be one of the key factors in the regulation of $\alpha3\beta4$ nAChRs in rat MPG neurons, which may play an important roles in the autonomic neuronal function such as synaptic transmission, autonomic reflex, and neuronal plasticity.

• 한국발생생물학회지:발생과생식
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• 제4권2호
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• pp.215-220
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• 2000

대부분의 내분비교란물질들은 에스트로겐이나 항에스트로겐적인 활성을 가지고 있어 사람이나 생태계에 있어서 생식기능 발달을 저해하는 것으로 보고되고 있다. 본 연구는 estrogen (E$_2$)과 bisphenol (BP) 그리고 octylphenol (OP)이 생쥐 Leydig cell line인 TM3 세포에 미치는 영향을 알아보고자 하였다. TM3 세포는 11 ~13일령의 BALB/c nu/＋ 생쥐로부터 유래한 정소내 세포인 Leydig cell로 DMEM에 FBS (10%)가 첨가된 배양액에 E$_2$, BP와 OP를 농도별 (1 pM, 1 nM, 1 $\mu$M, 1 mM)로 처리하고 48시간동안 체외에서 배양하였다. 배양 후, 전체 세포수와 생존율을 혈구세포판과 trypan blue 염색 방법으로 조사하였고, 스테로이드호르몬 합성에 관여하는 cytochrome P450scc (CYPscc)와 estrogen receptor $\alpha$ (ER $\alpha$) 유전자의 발현은 역전사 중합효소 연쇄 반응으로 관찰하였다. 결과를 살펴보면 TM3 세포의 생존율에 있어서는 1 $\mu$M 이하에서는 차이 가 없었으며 1 mM 첨가군에서는 유의하게 감소함을 보였다. 세포수에 있어서는 OP 처리군에서만 유의하게 적게 나타났다. CYPscc 유전자의 발현은 E$_2$ 군을 제외하고 BP (1 nM, 1 $\mu$M) 첨가군에서 약간 감소가, OP (1 nM, 1 $\mu$M) 첨가군에서는 유의한 감소가 나타났다. 그러나ER $\alpha$ 유전자의 발현은 처리군 모두에서 대조군보다 높은 발현율을 나타내었다. 결론적으로 고농도의 BP와 OP는 CYPscc유전자의 발현을 감소시킴으로써 스테로이드합성 과정을 억제시켜 정소기능에 장애를 일으키며, 정자형성 과정에도 영향을 미칠 수 있을 것으로 사료된다.

• 한국가축번식학회지
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• 제26권2호
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• pp.183-192
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• 2002

본 연구는 임신 및 분만후 rat에 oxytocin을 투여후 혈장, 자궁 및 태반 조직에서 estradiol-17$\beta$, progesterone, prostaglandin F$_2$$_{\alpha}$ 및 prostaglandin E$_2$그리고 자궁 조직의 oxytocin 수용체의 함량 사이의 관계에 대하여 조사하였다. 임신 rat에 oxytocin 투여 후 혈장 estradiol-17$\beta$의 농도는 임신 말기에 서서히 증가하여 임신 22일에 최고를 기록하였고 분만 후 1일에는 임신 16일 수준으로 감소하였다. Progesterone 농도는 임신 18일부터 분만 후 1일에 유의성 있는(p<0.05) 감소를 보였다. Prostaglandin F$_2$$_{\alpha}$ 및 Prostaglandin E$_2$농도는 임신 14일에서 임신 22일에 공히 상승하여 임신 22일에 최고를 기록하였고 분만 후 1일에는 급격히 감소하였다. Estradiol-17$\beta$의 농도는 자궁 조직에서 임신 14일에서 20일까지 서서히 증가하다가 임신 22일에 급격한 증가를 기록하였고 분만 후 1일에는 급격히 감소하여 임신 14일 수준을 유지하였으며 progesterone 농도는 임신 16일에 상승을 보인 후 임신 20일까지 급격히 감소하였고 분만 후 1일까지 같은 수준을 유지하였다. Prostaglandin F$_2$$_{\alpha}$ 및 prostaglandin E$_2$농도는 자궁조직에서 임신 말기 전기간에 상승하여 임신 22일에 최고를 기록한 후 분만 후 1일에는 급격히 감소하여 임신 14일과 비교하여 70% 수준을 기록하였다. Prostaglandin F$_2$$_{\alpha}$ 농도는 태반 조직에서 임신 14일에서 임신 22일까지 지속적으로 증가하였고 분만 후 1일에는 급격히 감소하였으며 prostaglandin E$_2$의 농도는 임신 14일에서 20일에 증가한 후 분만 후 1일까지 감소하였다. Oxytocin 수용체의 농도는 자궁 조직에서 임신 20일에서 임신 22일 사이에 급격히 증가한 후 분만 후 1일까지 비슷한 수준을 기록하였다. 이상에서와 oxytocin투여에 의한 임신 rat에 있어서 progesterone 농도의 감소에 이은 estradiol-17 $\beta$, prostaglandin F$_2$$_{\alpha}$, prostaglandin E$_2$및 oxytocin 수용체 농도의 증가는 조기에 시작되었다.

• 대한약리학회지
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• 제13권2호
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• pp.49-59
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• 1977

Adrenergic receptors are now classified into alpha type and beta type These adrenergic receptors are distributed in various tissue in different patterns. Therefore, the adrenergic response of a certain tissue may be different from those of the other tissues, and such differences may exist among various species of animals. In this paper, the authors attempt to reevaluate the effect of epinephrine on the isolated atria, aortic strips, and vas deferenses of rabbits preincubated with alpha receptor blockades (ergotamine and dibenamine) and beta receptor blockades (propranolol and dichloroisoproterenol) in Locke-Ringer bathing medium. The results obtained were summarized as follows; 1) The dose dependent responses of isolated atria to epinephrine were significantly inhibited by propranolol and dichloroisoproterenol, and slightly inhibited by dibenamine, but not affected by ergotamine. 2) The dose dependent responses of excised aortic strips to epinephrine were significantly inhibited by ergotamine and dibenamine, but the responses were slightly potentiated by propranolol, and significantly by dichloroisoproterenol. 3) The dose dependent responses of isolated vas deferenses to epinephrine were significantly inhibited by ergotamine and dibenamine, but slightly potentiated by propranolol and dichloroisoproterenol.

• 한방비만학회지
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• 제16권2호
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• pp.109-115
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• 2016

Objectives: The all anti-obesity drugs currently approved by the US Food and Drug Administration work by reducing energy intake. In fact, no approved drug targets energy expenditure. In Korean medicine, it is known to Qi or Yang invigorating therapy could increase energy metabolism. Aconitum carmichaeli Debx (ACD) is a Yang invigorating herb, often used for treat obesity in Korean medicine. In the present study, the authors investigated the regulatory effects of ACD in energy metabolism and mitochondrial biogenesis in C2C12 skeletal muscle cells. Methods: The water extract of ACD (0.2, 0.5 and 1.0 mg/ml) were treated in differentiated C2C12 cells. The protein or mRNA levels of target genes were analyzed and mitochondrial mass were investigated. Results: ACD activated the expressions of peroxisome proliferator-activated receptor gamma coactivator 1-alpha ($PGC-1{\alpha}$), nuclear respiratory factor 1 and TFAM and increased mitochondrial mass. ACD also increased adenosin monophosphate-activated protein kinase (AMPK), and acetyl-CoA carboxylase. Conclusions: This study suggests that ACD has the potential to increase energy metabolism and mitochondrial biogenesis by activating AMPK and $PGC1{\alpha}$.

• Journal of Microbiology and Biotechnology
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• 제25권2호
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• pp.247-254
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• 2015

In this presentation, I describe the expression and purification of the recombinant liver X receptor β-ligand binding domain proteins in E. coli using a commercially available double cistronic vector, pACYCDuet-1, to express the receptor heterodimer in a single cell as the soluble form. I describe here the expression and characterization of a biologically active heterodimer composed of the liver X receptor β-ligand binding domain and retinoid X receptor α-ligand binding domain. Although many of these proteins were previously seen to be produced in E. coli as insoluble aggregates or "inclusion bodies", I show here that as a form of heterodimer they can be made in soluble forms that are biologically active. This suggests that co-expression of the liver X receptor β-ligand binding domain with its binding partner improves the solubility of the complex and probably assists in their correct folding, thereby functioning as a type of molecular chaperone.

• Reproductive and Developmental Biology
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• 제39권1호
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• pp.1-6
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• 2015

Luteolysis is a cyclical regression of the corpus luteum in many non-primate mammalian species. Prostaglandin $F_2{\alpha}$($PGF_2{\alpha}$) from the uterus and ovary induces functional and structural luteolysis in bovine. The action of $PGF_2{\alpha}$ is mediated by $PGF_2{\alpha}$ receptor located on the luteal steroidogenic and endothelial cell membranes. $PGF_2{\alpha}$ plays an important role in regulating nitric oxide production in endothelial cells of the bovine corpus luteum. Nitric oxide production and nitric oxide synthase activity are stimulated and induced by $PGF_2{\alpha}$ in luteal endothelial cells. Moreover, the reactive oxygen species inhibits progesterone secretion in bovine luteal cells and induces apoptosis. Thus, the interaction between $PGF_2{\alpha}$ and reactive oxygen species provides important aspects in physiology of the corpus luteum forfunctional and structural luteolysis.

• The Korean Journal of Physiology and Pharmacology
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• 제21권5호
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• pp.495-507
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• 2017

The effect of clonidine administered intrathecally (i.t.) on the mortality and the blood glucose level induced by sepsis was examined in mice. To produce sepsis, the mixture of D-galactosamine (GaLN; 0.6 g/10 ml)/lipopolysaccharide (LPS; $27{\mu}g/27{\mu}l$) was treated intraperitoneally (i.p.). The i.t. pretreatment with clonidine ($5{\mu}g/5{\mu}l$) increased the blood glucose level and attenuated mortality induced by sepsis in a dose-dependent manner. The i.t. post-treatment with clonidine up to 3 h caused an elevation of the blood glucose level and protected sepsis-induced mortality, whereas clonidine post-treated at 6, 9, or 12 h did not affect. The pre-treatment with oral D-glucose for 30 min prior to i.t. post-treatment (6 h) with clonidine did not rescue sepsis-induced mortality. In addition, i.t. pretreatment with pertussis toxin (PTX) reduced clonidine-induced protection against mortality and clonidine-induced hyperglycemia, suggesting that protective effect against sepsis-induced mortality seems to be mediated via activating PTX-sensitive G-proteins in the spinal cord. Moreover, pretreatment with clonidine attenuated the plasma tumor necrosis factor ${\alpha}$ ($TNF-{\alpha}$) induced by sepsis. Clonidine administered i.t. or i.p. increased $p-AMPK{\alpha}1$ and $p-AMPK{\alpha}2$, but decreased p-Tyk2 and p-mTOR levels in both control and sepsis groups, suggesting that the up-regulations of $p-AMPK{\alpha}1$ and $p-AMPK{\alpha}2$, or down-regulations of p-mTOR and p-Tyk2 may play critical roles for the protective effect of clonidine against sepsis-induced mortality.