• The Korean Journal of Physiology
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• 제24권2호
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• pp.345-352
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• 1990

To determine the role of sulfhydryl group in transport of organic ions across the basolateral membrane of renal proximal tubules, effect of p-chloromercuribenzoic acid (PCMB) on the transport of tetraethylammonium (TEA) and p-aminohippurate (PAH) was studied in rabbit renal cortical slices. PCMB caused irreversible inhibition of TEA and PAH uptake in a dose-dependent manner, with $I_{50}$ value (concentration for 50% inhibition) of $30\;{\mu}M$ for TEA and $75\;{\mu}M$ for PAH. Kinetic analysis of TEA and PAH uptakes showed that PCMB decreased Vmax $(62.35\;vs.\;28.32\;n\;mole/g{\cdot}min\;fur\;TEA:\;385.24\;vs.\;170.36\;n\;mole/g{\cdot}min\;for\;PAH)$ without changing Km. The inhibitory action of PCMB on TEA and PAH uptakes was independent of pH of the pretreatment medium. The inhibitory effect of PCMB on the uptake of TEA or PAH was prevented by dithiothreitol, but not by the substrate. PCMB inhibited Na-K-ATPase activity in a dose-dependent manner with $I_{50}$ value of $50\;{\mu}M$, which is similar to those for TEA and PAH uptake. These results suggest that PCMB inhibits the transport of organic cations and anions in the renal basolateral membrane by directly affecting the SH-group in the transporter molecules or secondly by altering the Na-K-ATPase activity.

• The Korean Journal of Physiology
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• 제27권2호
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• pp.227-232
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• 1993

The temperature dependence of $Na^+-dependent$ succinate uptake was studied in brush border (BBMV) and basolateral (BLMV) membrane vesicles isolated from the rabbit kidney cortex. The succinate uptake was markedly altered by temperature in a similar fashion in both membranes. The temperature dependence was characterized by a nonlinear Arrhenius plot with a break point at 22 and $25^{\circ}C$ for BBMV and BLMV, respectively. The activation energy was 3.91 and 17.09 kcal/mole at above and below the break point respectively, far BBMV; 2.65 and 14.05 kcal/mole, respectively, for BLMV. When temperature increased f개m 20 to $35^{\circ}C$, the Vmax of succinate transport increased from $3.49{\pm}0.11\;to\;5.90{\pm}0.86\;nmole/mg/5\;sec$ for BBMV and from $2.86{\pm}0.25\;to\;3.63{\pm}0.32\;nmole/mg/5\;sec$ for BLMV, with no change in Km in both membranes. These results suggest that renal dicarboxylate transport is similarly sensitive to a change in membrane physical state in BBMV and BLMV.

• Parasites, Hosts and Diseases
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• 제28권4호
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• pp.197-206
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• 1990

MDCK세포간에 형성된 tight junction이 Toxoplasma gondii이 숙주세포 침투에 미치는 영향에 대하여 고찰 하기 위해 여러 조건의 세포배양을 시행하였다. MDCK세포를 25-well배양기 내의 18-mm cover glass에 $1{\times}10^5,{\;}3{\times}10^5{\;}및{\;}5{\times}10^5$ 개로 분주한 후 배양 1, 2, 3 및 4일에 다수의 Toxoplasma를 첨가하여 1시간 동안 배양한 다음 Giemsa 용액으로 염색한 후 관찰하였을 때, 각 실험군에서 침투한 원충의 숫자는 숙주세포가 포화밀도를 이루면서 급격히 감소하였다. 배양액 내의 calcium 농도를 일반적인 1.8mM에서 $5{\;}{\mu}M$로 낮추어 포화밀도의 MDCK 세포간에 tight junction 형성을 억제하였을 때, 원충의 침투가 약 2배 증가하였으며(p<0.05) 포화밀도 이전의 배양에서는 원충의 침투가 감소하였다. Trypsin-EDTA를 처리하여 포화밀도의 배양에서 tight junction을 소화 시킨 경우, 원충의 침투가 약 2.5배 증가하였으며 (p<0.05) 포화밀도 이전의 배양에서는 급격히 감소하였다. 이상의 결과들로 볼 때, 포화밀도의 MDCK세포간에 형성된 tight junction이 Toxoplasma의 침투를 억제하는 것을 알 수 있었다. 이는 Toxoplasma가 숙주세포로 침투할 때 숙주세포의 막구조에 대한 특이성이 있음을 시사 하며, 상피세포에서는 tight junction 위의 막(apical membrane) 보다 옆 및 아래의 막(basolateral membrane)상의 구조를 이용하는 것으로 추정되었다.

• 대한의생명과학회지
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• 제14권4호
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• pp.203-210
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• 2008

One of cell surface receptor proteins, human folate receptor (hFR) involves in the uptake of folates through cell membrane into cytoplasm, and is anchored to the plasma membrane by a fatty acid linkage, which has been identified in some cells as a glycosylphosphatidylinositol (GPI)-tailed protein with a molecular mass of about 40 kDa. The hFR is released by phosphatidylinositol phospholipase C (PI-PLC) because it contains fatty acids and inositol on the GPI tail. Caveolin decorates the cytoplasmic surface of caveolae and has been proposed to have a structural role in maintaining caveolae. It is unknown whether caveolin is involved in targeting, and is necessary for the function of GPI-tailed proteins. To compare the ability of folic acid binding, internalization and expression of hFR, and the effect of caveolin at the both apical and basolateral side of cell surfaces in Chinese hamster ovary (CHO) clone cells overexpressed the hFR and/or caveolin. Our present results suggest a possibility that the overexpression of caveolin does not be involved in expression of hFR, but plays a role as a factor in PI-PLC releasing kinetics, and for a regulation of formation, processing and function of hFR in CHO clone cells overexpressed cavcolin.

• Biomolecules & Therapeutics
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• 제10권4호
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• pp.281-286
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• 2002

In this study, we intended to get a preliminary data for establishing rat tracheal surface epithelial(RTSE) cell culture system as an experimental model for physiology and pharmacology of tracheal epithelial cells. Primary culture on the membrane support and application of the air-liquid interface system at the level of cell layer were performed. The cell growth rate and mucin production rate were measured according to the days in culture. The results were as follows: this culture system was found to manifest mucocilliary differentiation of rat tracheal epithelial cells, the cells were confluent and the quantity of produced and released mucin was highest on culture day 9, the mucin was mainly released to the apical side and tbe free $^3{H}$-glucosamine which was not incorporated to process of synthesis of mucin was left on the basolateral side. Taken together, we suggest that air-liquid interface culture system can be used as a substitute for immersion culture system and as an experimental model for in vivo mucus-hypersecretory diseases.

• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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• pp.234.3-235
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• 2003

The aim of this study is to evaluate the permeability of PEG-conjugated salmon calcitonin (sCT) across monolayers of Caco-2 cells that represent a model of the intestinal barrier. Caco-2 cells were grown to confluency on a permeable polycarbonate membrane to permit transport through it. Permeability experiments were performed with native-sCT and PEG-conjugated sCT (PEG M.W. 2000) at various concentrations (5uM, 10uM, 25uM, 50uM, 100uM) in the apical to basolateral direction. The barrier properties were assessed by detecting transport of markder molecules ($^3$H-mannitol) and by measuring transepithelial electrical resistance (TEER). (omitted)

• 한국발생생물학회지:발생과생식
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• 제22권3호
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• pp.263-273
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• 2018

Aquaporin (AQP) 3, a facilitated transporter of water and glycerol, expresses in placenta and fetal membranes, but the detailed localization and function of AQP3 in placenta remain unclear. To elucidate a role of AQP3 in placenta, we defined the expression and cellular localization of AQP3 in placenta and fetal membranes, and investigated the structural and functional differences between wild-type and AQP3 null mice. Gestational sacs were removed during mid-gestational period and amniotic fluid was aspirated for measurements of volume and composition. Fetuses with attached placenta and fetal membranes were weighed and processed for histological assessment. AQP3 strongly expressed in basolateral membrane of visceral yolk sac cells of fetal membrane, the syncytiotrophoblasts of the labyrinthine placenta and fetal nucleated red blood cell membrane. Mice lacking AQP3 did not exhibit a significant defect in differentiation of trophoblast stem cells and normal placentation. However, AQP3 null fetuses were smaller than their control litter mates in spite of a decrease in litter size. The total amniotic fluid volume per gestational sac was reduced, but the amniotic fluid-to-fetal weight ratio was increased in AQP3 null mice compared with wild-type mice. Glycerol, free fatty acid and triglyceride levels in amniotic fluid of AQP3 null mice were significantly reduced, whereas lactate level increased when compared to those of wild-type mice. These results suggest a role for AQP3 in supplying nutrients from yolk sac and maternal blood to developing fetus by facilitating transport of glycerol in addition to water, and its implication for the fetal growth in utero.

• The Korean Journal of Physiology
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• 제28권2호
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• pp.203-214
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• 1994

Using the methods of stopped-flow and epifluorescence microscopy with entrapped fluorophore, membrane permeability of $NH_3$ and weak acids in liposomes, renal brush border (BBMV) and basolateral membrane vesicles (BLMV), and primary culture cells from renal proximal tubule was measured. Permeability coefficient (cm/sec) of $NH_3$ was $(2.9{\times}10^{-2}$ in phosphatidylcholine liposome $25^{\circ}C)$, $5.9{\times}10^{-2}$ in renal proximal tubule cell $(37^{\circ}C)$, $4.0{\times}10^{-2}\;and\;2.4{\times}10^{-2}$ in BBMV and BLMV $(25^{\circ}C)$, respectively. Formic acid has the highest permeability coefficient among the weak acids tested, which was $4.9{\times}10^{-3}$ in liposome, $5.0{\times}10^{-3}$ in renal proximal tubule cell, $9.1{\times}10^{-3}$ in BBMV and $3.8{\times}10^{-3}$ in BLMV. There was a linear relationship between external concentration of nonionized formic acid and initial rate of flux of formic acid in liposome, and the slope coincided with the value of permeability coefficient of formic acid measured in pH 7.0. These results show that techniques of stopped-flow and epifluorescence microscopy with entrapped fluorophore provide the precise method of measurement of very rapid transport of nonelectrolytes through membranes with the advantages of instantaneous mixing effect, good resolution time and easy manipulation.

• Applied Microscopy
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• 제22권2호
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• pp.84-96
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• 1992

Turtle bladder 상피세포(上皮細胞)의 수송기작(輸送機作)을 in vitro에서 효과적으로 연구하기 위하여 Lucite chamber 한가운데 상피조직을 두고 전압고정법(電壓固定法)을 적용하여 상피 세포층의 막전위(膜電位)를 측정한 후 급속 동결(凍結)하고 투과 및 주사형 전자현미경(電子顯微鏡)으로 탄산 탈수효소를 함유하는 세포의 표면막 특성을 분석(分析)하였다. 방광(膀胱)의 점막층(粘膜層)은 두 타입의 탄산탈수효소를 함유한 세포가 특징적인데 정단부(丁端部)와 기저부(基底部) 세포막에서는 각기 다른 수송의 특성을 나타내고 있다. 즉 ${\alpha}$ 및 ${\beta}$형 탄산탈수효소가 풍부한 세포는 정단세포막(丁端細胞膜)의 proton 펌프를 이용하여 $H^+$ 분필(分泌)에 관여하거나 bicarbonate의 재흡수(再吸收) 기능을 가지는 것으로 믿어진다. 본 연구에서 탄산탈수효소를 함유한 ${\alpha}$형의 세포의 proton 분필수송(分泌輸送)과 세포막 투과성 변화와의 상관관계를 관찰하였는 바, 이들 세포에서 $H^+$을 분비하는 과정에서 정단부의 표면세포막(表面細胞膜) P-face에는 특이한 구조로서 세포막내(細胞膜內) 입자(粒子)들이 다량으로 분포하였다. 이와같은 세포막내(細胞膜內) 입자(粒子)들은 proton 펌프를 함유하는 것으로 생각되며 ${\beta}$형의 세포에서는 기저세포막(基底細胞膜)에서 관찰되고 있다. 이와같은 결과는 방광상피(膀胱上皮) 세포내 탄산탈수효소는 $H^+$과 $HCO_{3}^{-}$의 생성에 관여하지만, 특히 ${\alpha}$형 세포에서 정단세포막의 proton 펌프를 이용한 $H^{+}$ 분필수송(分泌輸送)과 기저세포막을 통한 bicarbonate의 재흡수(再吸收) 기능을 설명해 주는 중요한 사실로서 사료된다.

• 한국임상약학회지
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• 제23권4호
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• pp.316-321
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• 2013

Purpose: Atorvastatin, a HMG-CoA reductase inhibitor is widely prescribed in hyperlipidemic patients and telmisartan, an angiotensin receptor blocker is frequently used in the treatment of hypertension. Both drugs are substrates of organic anion transporting polypeptide (OATP) expressed in basolateral membrane in the liver, and undergo high first pass metabolism. Therefore, OATP-mediated hepatic uptake is important for disposition and metabolism of these drugs. The present study was designed to investigate the pharmacokinetic interactions between atorvastatin and telmisartan in rats. Method: Young adult SD rats were divided into three groups (n=6, each) and atorvastatin (10 mg/kg) and telmisartan (4 mg/kg) were orally given alone and together. Heparinized blood was serially taken and plasma concentrations of both drugs were measured using HPLC-MS/MS. Pharmacokinetic parameters of two drugs were calculated. Results: No significant pharmacokinetic change was found except a delay of time to peak of telmisartan when administered with atorvastatin. Each drug at the present dosage seemed to be insufficient to alter the pharmacokinetic parameters of its counterpart drug. Conclusion: Conclusively, co-administration of atorvastatin and telmisartan may lead to negligible clinical consequences.