• 대한수의학회지
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• 제47권4호
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• pp.371-378
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• 2007

To understand the role of $PM_{2X}/P_{2Y}$ receptor in cortex region of kidney and renal artery, molecular and functional analysis of $PM_{2X}/P_{2Y}$ receptor by pharmacophysiological skill in conventional swine tissues were performed. In functional analysis of $P_{2Y}$ receptor for vascular relaxation, 2-methylthio adenosine triphosphate, a strong agonist of $P_{2Y}$ receptor, induced relaxation of noradrenaline (NA)-precontracted renal artery in a dose-dependent manner. Strikingly, relaxative effect of ATP, 2-msATP, agonists of $P_{2Y}$ receptor, abolished by treatment of reactive blue 2, a putative $P_{2Y}$ receptor antagonist. In contrast, no significant differences of gene encoding $PM_{2X}/P_{2Y}$ and protein expression in immortalized suprachiasmatic nucleus from brain, primary isolated vascular smooth muscle cells from renal artery of pigs and HEK293 from human embryonic kidney under with/without adenosine triphosphate were observed. Taken together, the relationship between molecular and functional characteristic of $PM_{2X}/P_{2Y}$ receptors in conventional pig should be considered that they are another important factor which regulate the kidney function in swine. Based on this study, we propose the purinergic receptor as well as adrenergic and cholinergic receptors is an essential component of the renal homeostasis.

• Toxicological Research
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• 제8권2호
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• pp.343-357
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• 1992

Tumor necrosis factor-${\alpha}$(TNF), a polypeptide hormone secreted primarily by activated macrophages, was originally identified on the basis of its ability to cause hemorrhagic necrosis and tumor regression in vivo. Subsequently, TNF has been shown to be an important component of the host responses to infection and cancer and may mediate the wasting syndrome known as cachexia. These systemic actions of TNF are reflected in its diverse effects on target cells in vitro. TNF initiates its diverse cellular actions by binding to specific cell surface receptors. Although TNF receptors have been identified on most of animal cells, regulation of these receptors and the mechanisms which transduce TNF receptor binding into cellular responses are not well understood. Therefore, in the present study, the mechanisms how TNF receptors are being regulated and how TNF receptor binding is being transduced into cellular responses were investigated in rat liver plasma membranes (PM) and ME-180 human cervical carcinoma cell lines. $^{125}I$-TNF bound to high ($K_d=1.51{\pm}0.35nM$)affinity receptors in rat liver PM. Solubilization of PM with 1% Triton X-100 increased both high affinity (from $0.33{\pm}0.04\;to\;1.67{\pm}0.05$ pmoles/mg protein) and low affinity (from $1.92{\pm}0.16\;to\;7.57{\pm}0.50$ pmoles/mg protein) TNF binding without affecting the affinities for TNF, suggesting the presence of a large latent pool of TNF receptors. Affinity labeling of receptors whether from PM or solubilized PM resulted in cross-linking of $^{125}I$-TNF into $M_r$ 130 kDa, 90 kDa and 66kDa complexes. Thus, the properties of the latent TNF receptors were similar to those initially accessible to TNF. To determine if exposure of latent receptors is regulated by TNF, $^{125}I$-TNF binding to control and TNF-pretreated membranes were assayed. Specific binding was increased by pretreatment with TNF (P<0.05), demonstrating that hepatic PM contains latent TNF receptors whose exposure is promoted by TNF. Homologous up-regulation of TNF receptors may, in part, be responsible for sustained hepatic responsiveness during chronic exposure to TNF. As a next step, the post-receptor events induced by TNF were examined. Although the signal transduction pathways for TNF have not been delineated clearly, the actions of many other hormones are mediated by the reversible phosphorylation of specific enzymes or target proteins. The present study demonstrated that TNF induces phosphorylation of 28 kDa protein (p28). Two dimensional soidum dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) resolved the 28kDa phosphoprotein into two isoforms having pIs of 6.2 and 6.1. The pIs and relative molecular weight of p28 were consistent with those of a previously characterized mRNA cap binding protein. mRNA cap binding proteins are a class of translation initiation factors that recognize the 7-methylguanosine cap structure found on the 5' end of eukaryotic mRNAs. In vitro, these proteins are defined by their specific elution from affinity columns composed of 7-methylguanosine 5'-triphosphate($m^7$GTP)-Sepharose. Affinity purification of mRNA cap binding proteins from control and TNF treated ME-180 cells proved that TNF rapidly stimulates phosphorylation of an mRNA cap binding protein. Phosphorylation occurred in several cell types that are important in vitro models of TNF action. The mRNA cap binding protein phosphorylated in response to TNF treatment was purifice, sequenced, and identified as the proto-oncogene product eukaryotic initiation factor-4E(eIF-4E). These data show that phosphorylation of a key component of the cellular translational machinery is a common early event in the diverse cellular actions of TNF.

• Clinical and Experimental Pediatrics
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• 제52권7호
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• pp.811-817
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• 2009

목 적 : 소아 천식에서 흡입용 스테로이드의 장기간 사용이 골밀도와 골대사에 미치는 영향에 대해 알아보고자 하였다. 방 법 : 고려대학교 안암병원 소아청소년과에서 천식으로 진단되어 최소 6개월 이상 ICS를 사용 중인 만 6세에서 12세 사이의 환아(ICS군: 26명)를 대상으로 하였다. 대조군으로 연령 및 성별이 일치하면서 LTRA만을 사용한 천식 환아(LTRA군: 15명)와 골밀도에 영향을 미칠 만한 기저 질환이 없는 정상 소아(정상군: 30명)를 선정하여 이중 에너지 X-선 흡수법으로 요추와 대퇴골 상부에서 골밀도를 측정하고, 골대사에 대한 영향은 혈청 골-특이 알칼리인산효소(BALP)를 측정하여 평가하였다. 결 과 : ICS군의 요추 골밀도는 $0.57{\pm}0.07g/cm^2$으로 LTRA군($0.55{\pm}0.06g/cm^2$) 및 정상군($0.58{\pm}0.07g/cm^2$)과 유의한 차이를 보이지 않았고(P=0.254), 대퇴골 상부의 골밀도도 ICS군에서 $0.70{\pm}0.07g/cm^2$로 LTRA군($0.66{\pm}0.06g/cm^2$)및 정상군($0.70{\pm}0.07g/cm^2$)과 유의한 차이가 없었다(P=0.297). 골밀도 Z-점수 또한 세 군에서 유의한 차이를 보이지 않았다(요추: ICS군 $-0.22{\pm}0.78$ vs. LTRA군 $-0.16{\pm}0.47$ vs. 정상군 $-0.19{\pm}0.83$, P=0.963; 대퇴골 상부: ICS 군 $-0.12{\pm}0.93$ vs. LTRA군 $0.16{\pm}0.59$ vs. 정상군 $-0.04{\pm}0.67$, P=0.560). 혈청 BALP 농도는 ICS군과 LTRA군에서 정상군에 비해 유의하게 높은 소견을 보였다(P=0.021). ICS군에서 ICS의 최근 6개월간 사용량, 총 축적 용량, 사용 기간, 투여 시작 연령과 골밀도 Z-점수는 유의한 상관관계를 보이지 않았다(모두 P>0.05). 결 론 : 천식 환아에서 저용량의 ICS 사용은 골밀도에 유의한 영향을 미치지 않음을 확인하여, ICS의 부작용에 대한 과도한 우려를 막고 치료 순응도를 높이는데 도움이 될 것으로 생각한다.

• Asian-Australasian Journal of Animal Sciences
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• 제20권4호
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• pp.569-576
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• 2007

Four hundred and fifty tilapias ($6.77{\pm}0.23$ g) were assigned randomly to six groups to evaluate the feasibility of the tested antibacterial peptides (ABPs) and oligosaccharides as substitutes for antibiotics. The control group was fed with a commercial tilapia diet; other five groups were fed with the same commercial diet supplemented with konjac glucomannan (KGLM), cluster bean galactomannan (CBGAM), and three animal intestinal ABPs derived from chicken, pig and rabbit at 100 mg/kg respectively. After 21 days of feeding, growth, disease resistance, and in vivo anti-adherence were determined. Furthermore, the inhibitory effect of tested agents on adhesion of Aeromonas veronii biovar sobria (A.vbs) strain BJCP-5 to tilapia enteric epithelia in vitro was assessed by cell-ELISA system. As a result, the tested agents supplemented at 100 mg/kg show significant benefit to tilapia growth and disease resistance (p<0.05), and the benefit may be correlated with their interfering in the contact of bacteria with host mucosal surface. Although none of the tested agents did inhibit the growth of BJCP-5 in tryptic soy broth at $100{\mu}g/ml$, all of them did inhibit the adhesion of A.vbs to tilapia enteric epithelia in vivo and in vitro. In vitro mimic assays show that three ABPs at low concentrations of $25{\mu}g/ml$ and $2.5{\mu}g/ml$ have the reciprocal dose-dependent anti-adherence effect. The inhibition of ABPs may be correlated with a cation bridging and/or receptor-ligand binding, but not with hydrophobicity. The KGLM and CBGAM inhibited the adherence of BJCP-5 to tilapia enteric epithelia with dose-dependent manner in vitro, and this may be through altering bacterial hydrophobicity and interfering with receptor-ligand binding. Our results indicate that the anti-adherence of the tested ABPs and oligosaccharides may be one of the mechanisms in promoting tilapia growth and resistance to A.vbs.