• The Korean Journal of Physiology
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• 제20권2호
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• pp.225-235
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• 1986

가토 근위세뇨관에서 Percoll gradient에 의해 분리한 기저막소포(basolateral membrane vesicle)에서 PAH 이동에 미치는 여러 음이온들의 영향을 rapid filtration technique을 이용하여 관찰하였다. NA-dependent PAH 축적은 용액내 $SO_4$와 $SSO_3$에 의하여 유의하게 억제되었으나 $Cl,\;SCN,\;HPO_4,\;acetate$ 및 oxalate에 의해서는 영향을 받지 않았다. 용액내의 $SO_4$는 상경적으로 PHA 이동을 억제하였다. 소포내 음이온을 부하시킨 후 PAH 이동을 측정했을 때 $acetate,\;SO_4$ 및 PAH의 부하에 의해 PAH 이동은 유의하게 증가되었으며, 소포내 부하되는 $SO_4$농도가 증가함에 따라 PAH 축적은 증가되었다. $SO_4$에 의한 PAH의 trans-stimulation은 Na 농도 경사 존재시 더욱 증가되었으며 이들은 2 mM probenecid 및 1mM SITS에 의해 억제되었다. 이들 결과들은 신피질세뇨관 세포의 기저막에 PAH/음이온 교환에 의해 PAH가 이동되는 기전이 존재한다는 것을 가르킨다. 그러나 PAH 이동에 영향이 없는 음이온들은 PAH/음이온 교환에 기질로써 작용하지 않기 때문인지 아니면 이들 음이온들의 높은 투과성에 인해 나타난 결과인지는 더욱 추구해 보아야 확인될 것으로 생각된다.

• The Korean Journal of Physiology
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• 제22권2호
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• pp.281-293
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• 1988

가토 신장 근위세뇨관에서 분리한 brush border membrane vesicle(BBMV)과 basolateral membrane vesicle(BBMV)에서 rapid filtration 방법으로 PAH 이동에 대한 pH의 영향을 관찰하였다. BLMV에서 용액내 Na이 없을 때 외부 $pH(pH_0)$를 8.0에서 5.5까지 감소시켰을 때 probenecid-sensitive PAH 이동은 유의하게 증가되었다. 용액내 Na이 있을 때 $pH_0$가 8.0에서 6.0까지 변화하여도 PAH 이동에는 영향이 없었으나 5.5까지 더욱 감소시켰을 때 PAH 이동이 증가하였다. 그러나 vesicle 내 외에 pH gradient없이 $pH_0$를 내부 $pH(pH_1)$와 동일하게 변화시켰을 때 PAH 이동은 영향을 받지 않았다. PH gradient가 없을 때 시험된 pH범위에서 Na은 PAH 이동을 증가시켰다. BBMV에서도 BLMV에서와 유사한 pH 의존성을 보였으나 Na의 존재는 PAH 이동에 영향을 미치지 못하였다. BLMV에서 동력학적 분석 결과 일정한 $pH(pH_1)$에서 $pH(pH_0)$ 감소는 Km에 변화없이 PAH 이동에 대한 Vmax를 유의하게 증가시켰다. 이러한 결과로 BBMV에서 PAH 이동에 대한 pH의 영향은$OH^-/PAH$교환기전에 기인하는 것으로 추측되나 BLMV에서 pH 의존성은 음이온 교환기전만으로 설명될 수 없다. 또한 BLMV에서는 PAH 이동이 Na에 의존하나 BBMV에서는 Na에 의존하지 않음을 가르킨다.

• 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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• 제21권2호
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• pp.283-289
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• 1987

Uric acid transport across the basolateral membrane of renal proximal tubules was studied in rabbit kidney cortical slices. Uric acid uptake was greater under $O_2$ atmosphere compared to under $N_2$ atmosphere, and was increased with $Na^{2+}$ concentration in incubation medium. Uric acid inhibited PAH uptake but not TEA uptake and did trans-stimulated PAH efflux. PAH also inhibited uric acid uptake. Uric acid uptake was inhibited by harmaline, ouabin, SITS, DIDS and pyrazinoic acid. The inhibition of PAH uptake by these inhibitors also was reasonably comparable to that of uric acid uptake. These results suggest that uric acid was transported across the basolateral membrane of renal tubule by a carrier-mediated process which was by a common transport system with PAH in rabbit.

• The Korean Journal of Physiology
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• 제19권1호
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• pp.49-59
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• 1985

The effects of anions on net accumulation of $(\rho-aminohippuric\;acid)$(PAH) were studied in rabbit kidney cortical slices. Experiments were carried while varying the major anionic composition of the incubation medium(replacement of $Cl^-$ by isethionate and $SCN^-$). The total replacement of $Cl^-$ with isethionate, $SO_4\;^{2-}$ and $SCN^-$ in the incubation medium decreased the 60-min slice-to-medium concentration(S/M) ratio of PAH to 60%, 40% and 50% of control value, respectively. The degree of inhibition in PAH accumulation by the replacement of isethionate and $SCN^-$ was increased with increasing of both preincubation and incubation time. The influence of isethionate and $SCN^-$ on PAH uptake was fully reversible. Both isethionate and $SCN^-$ increased the apparent Km value significantly with no change on the apparent Vmax value, suggesting a competitive inhibition on PAH uptake. And the inhibitory effect of $SCN^-$ on PAH uptake decreased with increase of pH in the incubation medium while that of isethionate increased with increase of pH. Intracellular water content, intracellular electrolyte concentration and oxygen consumption were not influenced by the replacement of $Cl^-$ with isethionate or $SCN^-$ in the incubation medium. These results suggest that both $isethionate^-$ and $SCN^-$ inhibit the PAH uptake by binding to some site necessary for normal PAH transport without affecting the cellular viability.

• The Korean Journal of Physiology
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• 제30권2호
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• pp.279-288
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• 1996

Chronic exposure to cadmium impairs various renal tubular functions, including organic acid (anion) secretion. To investigate the mechanism of cadmium-induced alterations in the organic anion transport system, kinetics of p-aminohippurate (PAH) uptake was studied in renal cortical basolateral membrane vesicles (BLMV) isolated from cadmium-intoxicated rats (adult male Sprague-Dawley). Cadmium intoxication was induced by subcutaneous injections of $CdCl_{2}$ (2 mg Cd/kg per day) for 3 weeks. The renal plasma membrane vesicles were prepared by Percoll gradient centrifugation. The vesicular uptake of $^{14}C$-PAH was determined by rapid filtration technique using Millipore filter. Cadmium intoxication resulted in a marked attenuation of $Na^{+}$-dependent, ${\alpha}$-ketoglutarate (${\alpha}$KG)-driven PAH uptake with no changes in $Na^{+}$ and ${\alpha}$KG-independent transport component. Kinetic analysis indicated that Vmax, but not Km, of the $Na^{+}$-dependent, ${\alpha}$KG-driven component was reduced. A similar reduction of $Na^{+}$-dependent, ${\alpha}$KG-driven PAH uptake was observed in normal membrane vesicles directly exposed to inorganic cadmium in vitro, and this was accompanied by an inhibition of both $Na^{+}$-dependent ${\alpha}$KG uptake and ${\alpha}$KG-PAH exchange activity. These results indicate that during chronic exposure to cadmium, free cadmium ions liberated in the proximal tubular cytoplasm directly interact with the basolateral membrane and impair the active transport capacity for organic anions, most likely due to an inhibition of both $Na^{+}$-dicarboxylate cotransporter and dicarboxylate-organic anion antiporter activities.

• 대한약침학회지
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• 제4권2호
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• pp.49-56
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• 2001

This study was undertaken to determine whether Scutellaria baicalensis Georgi (SbG) extract exerts the protective effect against $HgCl_2$-induced alterations in membrane transport function in rabbit renal cortical slices. The slices were treated with 0.1 mM $HgCl_2$ for 60 min at $37^{\circ}C$. $HgCl_2$ caused an inhibition in PAH uptake by renal cortical slices. Such an effect was accompanied by depressed $Na^+-K^+$-ATPase activity and ATP depletion. SbG prevented $HgCl_2$-induced inhibition of PAH uptake in a dose-dependent manner at the concentration ranges of 0.01-0.1%. $HgCl_2$-induced inhibition of $Na^+-K^+$-ATPase activity and ATP depletion were significantly prevented by 0.05% SbG. These results suggest that SbG prevents $HgCl_2$-induced alterations in membrane transport function in rabbit renal cortical slices. Such protective effects of SbG may be attributed to inhibition of peroxidation of membrane lipid.

• Journal of Acupuncture Research
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• 제18권3호
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• pp.123-133
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• 2001

Objective : This study was undertaken to determine whether Cordyceps sinensis Sacc. (CSS) extract exerts the protective effect against oxidant-induced alterations in membrane transport function in renal tubules. Methods : Membrane transport fucntion was estimated by examining alterations in p-aminohippurate (PAH) uptake in rabbit renal cortical slices. For induction oxidative stress, slices were treated with an organic peroxide cumene hydroperoxide for 60 min at $37^{\circ}C$. Cumene hydroperoxide inhibited PAH uptake in a time dependent manner. Results : CSS at 0.5-5% concentrations prevented cumene hydroperoxide-induced inhibition of PAH uptake. CSS at 1% also attenuated LDH release and lipid peroxidation induced by cumene hydroperoxide. When slices were treated with 0.2 mM mercury chloride, PAH uptake was inhibited and lipid peroxidation was increased. These changes by mercury were significantly prevented by CSS. Conclusion : These results suggest that CSS prevents oxidant-induced alterations in membrane transport function in rabbit renal cortical slices. Such protective effect of CSS may be attributed to inhibition of peroxidation of membrane lipid.

• The Korean Journal of Physiology
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• 제22권1호
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• pp.55-62
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• 1988

가토 신피질 절편에서 p-aminohippurate (PAH) 이동에 대한 카드뮴 이온(Cd)의 영향을 조사하여 다음과 같은 결과를 얻었다. 조직절편과 용액내의 PAH 농도비(S/M PAH)는 용액내에 Cd이 0.1 mM이상 존재할 때에 현저히 감소되었다. PAH influx에 대한 동력학적 분석결과 Cd에 의하여 influx의 최대이동율(즉 $V_{max}$)은 심하게 감소되지만 운반체와 기질간의 친화력(즉 $K_{m}$)에는 변화가 없었으며, 수동적 influx 역시 변화되지 않았다. 신피질조직의 산소 소모율은 1 mM Cd에 의하여 35% 가량 억제되었으며, 신피질 microsome 분획의 Na-K-ATPase 활성도는 Cd 농도가 $10^{-7}M$ 이상일 때 의의있게 억제되었다. 이상의 결과로 미루어 볼 때 신장조직이 카드뮴이온에 직접 노출될 경우 유기음이온의 능동적 이동능이 심하게 저해된다고 사료된다.

• The Korean Journal of Physiology
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• 제17권2호
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• pp.143-159
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• 1983

Vanadate가 가토 신피질절편에서 PAH이동과 Na-K-ATPase활성에 미치는 효과를 관찰한 결과 다음과 같은 결론을 얻었다. 1) Vanadate는 Na-K-ATPase활성을 농도에 따라 억제하였으며 $7.94{\times}10^{-7}M$에서 이 효소의 활성이 50% 억제되었다. 2) Vanadate는 PAH의 능동적이동을 농도에 따라 억제하였으며 50%억제농도는 대략 $10^{-4}M$ 이었고, 수동적이동에는 영향을 미치지 못하였다. 조직내 Na과 K의 양도 vanadate가 PAH이동을 억제하는 농도 범위에서 같이 변화하였고 산소소모량은 $10^{-4}M$까지는 약간 감소하였으나 $10^{-3}M$에서는 오히려 증가하였다. 3) 30분간 preincubation한 후에도 15분까지의 PAH이동은 30분 이후에 비해 vanadate에 의해 적게 억제되었다. 4) $10^{-4}M$ vanadate와 $10^{-4}M$ ouabain은 가역적으로 PAH 이동을 억제하였으며 $10^{-3}M$ vanadate는 비가역적으로 억제하였고 장시간 세척후에도 거의 같은 정도의 억제양상을 나타내었다. 5) Vanadate에 의한 PAH이동의 억제정도는 incubation용액내 $Na^+$의 감소, $K^+$의 증가에 의하여 증가하였고 $Ca^{2+}$의 농도 변화에 의해서는 영향을 받지 않았다. 6) Vanadate가 존재치 않을 때 Tris완충용액 사용시는 pH 8.2까지 PAH축적정도가 증가하였고 phosphate완충용액 사용시는 pH 7.4에서 최대축적치를 보였다. pH가 증가함에 따라 억제정도는 증가하였으며 같은 pH에서도 완충용액의 종류에 따라 vanadate에 의한 억제정도가 달랐다. 7) Vanadate와 ouabain은 PAH이등과 Na-K-ATPase활성에 부가적 억제작용을 나타내었다. 이상의 결과로 vanadate는 가토신장의 세포내부에서 Na-K-ATPase를 가역적으로 억제함으로써 PAH의 이동을 억제하는 것으로 생각되며 PAH의 이동은 Na-K-ATPase활성과 기능적으로 밀접히 연결되어 있는 것으로 생각된다.