• 한국환경보건학회지
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• 제19권4호
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• pp.83-89
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• 1993

To evaluate an effect of methanethiol on a cause of erythrocyte membrane damage in rats, methanethiol was given at 11.25 rag/100 g body weight, and after 4 hr, the animals were sacrifled, the activities of Na$^+$/K$^+$ ATPase, protein contents in partial purified erythrocyte membrane and erythrocyte indices were determined Concomitantly, in vitro, effect of methanethiol on the erythrocyte fragility, Na$^+$/K$^+$ ATPase activity and its kinetics in various concentration of substrate from the preincubated erythrocyte membrane with methanethiol were demonstrated. The spleen weight per body weight (%) and MCV of erythrocyte in methanethiol-treated rats were more increased than those in the control group. The Na$^+$/K$^+$ ATPase activities in erythrocyte membrane were more decreased in methanethiol-treated rats than those in the control group. The apply of 0.05 ng rat whole blood to the 0.24 mg/ng of methanethiol solution in isotonic condition showed the complete hemolysis. The Na$^+$/K$^+$ ATPase activity in preincubated erythrocyte membrane with methanethiol at 37$\circ$C showed the dual effect and the K$_m$ value of Na$^+$/K$^+$ ATPase was higher in the preincubated erythrocyte membrane with methanethiol than that in the preincubated erythrocyte membrane omitted the methanethiol. These results suggest that the methanethiol may induce the damage of rat's erythrocyte membrane due to a change in substrate binding affinity of Na$^+$/K$^+$ ATPase.

• The Korean Journal of Physiology
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• 제10권1호
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• pp.15-24
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• 1976

The action of aconite on the sodium plus potassium activated ATPase activity in the rabbit red cell membrane has been investigated and the experiments were also designed to determine the mechanism of action of aconite on the ATPase activity. The following results were observed. 1. The activity of the NaK ATPase from red cell membrane is stimulated by aconite, and the concentration of aconite for maximal activity is about 80 mg%. The pH optimum for the aconite sensitive component is 8.0. 2. The activating effect of aconite on the ATPase, with a given concentration of sodium in the medium, is increased by raising the potassium concentration but activity ratio is decreased. 3. The activating effect of aconite on the ATPase, with a given concentration of potassium in the medium, is increased by raising the sodium concentration but activity ratio is decreased. 4. The action of aconite on the ATPase activity is inhibited by calcium ions and the effect of inhibition is increased by small amounts of calcium but decreased by larger amounts. 5. The activating effect of aconite on the ATPase was not related to the sulfhydryl group of cysteine, the amino group of lysine, the hydroxyl group of threonine or the imidazole group of histidine. 6. The action of aconite on the ATPase activity is due to carboxyl group of the enzyme of NaK ATPase.

• The Korean Journal of Physiology
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• 제12권1_2호
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• pp.15-23
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• 1978

The action of ascorbic acid on the sodium Plus potassium activated ATPase activity in the rabbit red cell membrane has been investigated and the experiments were also designed to determine the mechanism of action if ascorbic acid on the ATPase activity The following results were observed. 1. The activity of the NaK ATPase from red cell membrane is stimulated by ascorbic acid and the concentration of ascorbic acid for maximal activity is about 8 mM. 2. The activating effect of ascorbic acid on the ATPase activaty, with a given concentration of sodium in the medium, is increased by raisins the potassium concentration but activity ratio is decreased. 3. The activating effect of ascorbic acid on the ATPase activity, with a given concentration of potassium in the medium, is increased by raising the sodium concentration but activity ratio is decreased. 4. The action of ascorbic acid on the ATPase activity is stimulated by calcium ions and activity ratio is increased by raising the calcium concentration. 5. The activating effect of ascorbic acid on the ATPase activity was not related to the sulfhydryl group of cysteine or the hydroxyl group of threonine. 6. The activating effect of ascorbic acid on the ATPase activity is due to amino group and carboxyl group of the enzyme of NaK ATPase.

• 대한약리학회지
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• 제16권1호
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• pp.51-56
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• 1980

Phytolaccae Radix (PR), Brunella Herba (BH), Akebiae Lignum (AL) and Atractylis Rhizoma (AR) are some of the diuretic agents used in Chinese medicine and folk remedy. Water or methanol extracts of them (100mg/kg) were intravenously injected to rabbits in order to re-evaluate the effects on renal function. PR water extract elicited moderate diuresis while water extracts of BH, AL and methanol extract of AR had antidiuretic effects. Influence of PR on renal hemodynamics and $Na^+-K^+$-ATPase activity in rabbit kidney were observed in vivo and in vitro. The results were as follows: 1) Clearances of inulin and p-aminohippuric acid increased significantly after 15 minutes following the administration of PR water extract, but Na+ reabsorption rate was not changed. 2) The increase of $Na^+-K^+$-ATPase activity in renal cortex, outer and inner medulla was observed at 15 minutes after PR water fraction was given intravenously, and the change was most prominent in cortical area. 3) More than 50% of decrease in $Na^+-K^+$-ATPase activity in renal tissues was observed with PR water fraction $(10^{-2}g/ml)$ in vitro experiments. However, the inhibition of $Na^+-K^+$-ATPase activity was reversed with lower concentrations $(10^{-4}g/ml,\;10^{-6}g/ml)$ of PR water fraction in outer and inner medullary zone. These results suggest the diuretic effect of PR is due to improved renal hemodynamics, and contradictory reults concerning $Na^+-K^+$-ATPase activity require further investigation.

• The Korean Journal of Physiology
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• 제19권1호
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• pp.15-23
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• 1985

$(Na^++K^+)-ATPase$은 ${\alpha}$ 와 ${\beta}$의 두 subunits로 구성되어 있으며, 분자량이 약 300,000 daltons 정도되는 것으로 보아 ${\alpha}_2{\beta}_2$의 형태로 존재할 것으로 알려져 왔다 한편, 사람 적혈구막에 있는 $Na^+,\;K^+\;Pump$는 glycolytic enzymes과 complex를 이루고 있으리라는 보고도 있다. 우리는 이 실험에서 in situ상태의 사람 적혈구막$(Na^++K^+)-ATPase$의 분자량을 측정하기 위하여, 소위 말하는 ‘Target theory’를 radiation에 의한 ouabain sensitive한 $\Na^+$이동과, intact한 cells과 ghosts에서의 ATP가수분해능력의 inactivation data에 적용하였다. Intact한 cells은 cryoprotective agent의 존재하에서, ghosts는 직접적으로 액화질소의 용기속에 담고 온도를 $-45^{\circ}C$에서 $-50^{\circ}C$로 유지시키면서 1.5 MeV의 electron beam으로 조사한 후에 Pump의 기능내지 효소의 활성도를 측정하여 radiation에 따르는 inactivation의 정도를 측정하였다. 이득 활성도는 radiation의 양에 따라 simple exponential function으로 inactivation되었으며, 이로부터 radiation sensitive volume(target size)를 계산하였다. Target size는 intact한 cells을 사용하였을 경 우$(Na^++K^+)-ATPase$나 $Na^+,\;K^+\;Pump$ 모두 600,000 daltons으로 계산되었으며, 이 값은 만약 cells을 strophanthidin으로 먼저 처치하고 측정하면 약 325,000 daltons으로 감소하였다. Ghosts를 사용했을 경우에도$(Na^++K^+)-ATPase$의 target size는 역시 약 325,000 daltons이었다. 이상의 결과로 미루어 보아 intact한 cells에서는 $(Na^++K^+)-ATPase/Na^+,\;K^+\;Pump$가 $(\alpha\beta)_2$의 dimer 상태로 존재하거나 혹은 $(\alpha\beta)_2$의 monomer에 glycolytic enzymes과 같은 다른 enzymes이 붙어 functional한 구조를 이루고 있는 것이 아닌가 사료된다. 또한 실헐성적은 이러한 dimeric association 혹은 heterocomplex association은 ghost를 만드는 과정에서나 strophanthidin의 처치로 부서질 수 있음을 암시하고 있다.

• 대한약리학회지
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• 제2권1호
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• pp.31-40
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• 1966

The microsomal fraction is isolated from rabbit heart and skeletal muscle. The fraction is found to contain the $Na^+$-and $K^+$-activated ATPase. The maximal ATPase activity is obtained in $Na^+$ and $K^+$ concentration of 100 mM. Calcium itself stimulates the $Na^+$-and $K^+$-activated portion of ATPase in the presence of $Mg^{++}$. However, calcium does not stimulate ATPase in the absence of $Mg^{++}$.

• Toxicological Research
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• 제34권2호
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• pp.143-150
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• 2018

It has been demonstrated that vanadate causes nephrotoxicity. Vanadate inhibits renal sodium potassium adenosine triphosphatase (Na, K-ATPase) activity and this is more pronounced in injured renal tissues. Cardiac cyclic adenosine monophosphate (cAMP) is enhanced by vanadate, while increased cAMP suppresses Na, K-ATPase action in renal tubular cells. There are no in vivo data collectively demonstrating the effect of vanadate on renal cAMP levels; on the abundance of the alpha 1 isoform (${\alpha}_1$) of the Na, K-ATPase protein or its cellular localization; or on renal tissue injury. In this study, rats received a normal saline solution or vanadate (5 mg/kg BW) by intraperitoneal injection for 10 days. Levels of vanadium, cAMP, and malondialdehyde (MDA), a marker of lipid peroxidation were measured in renal tissues. Protein abundance and the localization of renal ${\alpha}_1-Na$, K-ATPase was determined by Western blot and immunohistochemistry, respectively. Renal tissue injury was examined by histological evaluation and renal function was assessed by blood biochemical parameters. Rats treated with vanadate had markedly increased vanadium levels in their plasma, urine, and renal tissues. Vanadate significantly induced renal cAMP and MDA accumulation, whereas the protein level of ${\alpha}_1-Na$, K-ATPase was suppressed. Vanadate caused renal damage, azotemia, hypokalemia, and hypophosphatemia. Fractional excretions of all studied electrolytes were increased with vanadate administration. These in vivo findings demonstrate that vanadate might suppress renal ${\alpha}_1-Na$, K-ATPase protein functionally by enhancing cAMP and structurally by augmenting lipid peroxidation.

• The Korean Journal of Physiology
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• 제23권1호
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• pp.23-34
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• 1989

Gentamicin (GM) is a polybasic, aminoglycoside antibiotic used frequently for the treatment of serious gram-negative infections. The major limiting factors in the clinical use of GM as well as other aminoglycoside antibiotics are their nephrotoxicity and ototoxicity. The primary mechanism of cell injury in aminoglycoside toxicity appears to be the disruption of normal membrane function and the inhibition of $Na^{+}-K^{+}$ ATPase activity. There are both indirect and direct evidences which suggests that the effect of aminoglycoside antibiotics on $Na^{+}-K^{+}$ ATPase may explain, or contribute to, their toxicity. It has been shown that aminoglycoside reduce total ATPase activity (Kaku et al., 1973) and $Na^{+}-K^{+}$ ATPase activity (linuma et al., 1967) in the stria vascularis and spiral ligament of the guinea-pig cochlea. Lipsky and Lietman (1980) reported that aminoglycoside antibitoics inhibited the activity of $Na^{+}-K^{+}$ ATPase in microsomal fractions of the cortex and medulla of the guinea-pig kidney, isolated rat renal tubule and human erythrocyte ghosts. The present invstigation was undertaken to elucidate the mechanism of GM on human erythrocytes by examining its effect on $Na^{+}-K^{+}$ ATPase activity, actives sodium and potassium transport across red blood cell and $^{3}H-ouabain$ binding to red blood cell membranes. The results obtained are summarized as follows: 1) CM inhibited significantly both the activity of total ATPase and $Na^{+}-K^{+}$ ATPase at all concentrations tested. 2) GM inhibited active $^{22}Na$ efflux across red blood cell. When ouabain is present, the rate of $^{22}Na$ efflux was completely inhibited. When both GM and ouabain were added, the inhibitory effect of active $^{22}Na$ efflux was more pronounced. 3) Active $^{86}Rb$ influx was inhibited significantly by GM. In the presence of ouabain, the rate of $^{86}Rb$ influx is markedly inhibited. But $^{86}Rb$ influx is not appreciably altered by the presence of both GM and ouabain. 4) In the presence of GM, $^{3}H-ouabain$ binding to red blood cell membrane increased. From the above results, it may be concluded that the inhibition of active sodium and potassium transport across red blood cell by gentamicin appears to be due to the inhibition of $Na^{+}-K^{+}$ ATPase activity and an increase in ouabain binding to red blood cell membranes.

• Radiation Oncology Journal
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• 제12권2호
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• pp.129-141
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• 1994

Intracellular ions which have a major role in cellular function have been reported to affect repair of radiation damage. Recently it has been reported that ouabain sensitizes A549 tumor cellls but not CCL-120 normal cells to radiation. Ouabain inhibits the $Na^+-K^+$-pump rapidly thus it increases intracellular Na concentration, Vanadate which is distributed extensively in almost all living organisms is known to be a $Na^+-K^+$-ATPase inhibitors, This study was performed to see any change in radiosensitivity of tumor cell by vanadate and any role of $Na^+-K^+$ATPase in radiosensitization. Experiments have been carried out by pretreatment with vanadate in human cell line(A549, JMG) and mouse cell line(L1210, spleen). For the cell survival MTT assay was performed for A549 and JMC cells and frypan blue dye exclusion test for L120, and spleen cells. Measurements of $Na^+-K^+$-ATPase activity in control, vanadate treated cell, radiation treated cell (9 Gy for A549 and JMG, 2 Gy for L1201, spleen), and combined $10^{-6}M$ vanadate and radiation treated cells were done. The results were summerized as fellows. 1. L1210 cell was most radiosensitive, and spleen cell and JMG cell were intermediate, and A549 cell was least radiosensitive. 2. Mininum or no cytotoxicity was seen with vanadate below concentration of $10^{-6}M$. 3. In A549 cells there was a little change in radiosensitivity with treatment of vanadate. However radiation sensitization was shown in low dose level of radiation i. e. 2- Gy. In JMG cells no change in radiosensitivity was noted. Both L1210 and spleen cell had radiosensitization but change was greater in tumor cell. 4. $Na^+-K^+$-ATPase activity was inhibited significantly in tumor cell by treatment of vanadate. 5. Radiaiton itself inhibited $Na^+-K^+$-ATPase activity of tumor cell with high $Na^+-K^+$-ATPase concention. Increase in radiosensitivity by vanadate was closely associated with orginal $Na^+-K^+$-ATPase contents. From the above results vanadate had little cytotoxicity and it sensitized tumor cells to radiation. Inhibitory effect of vanadate on $Na^+-K^+$-ATPase activity might be one of the contributing factors for radiosensitization to tumor cells which has greater enzyme activity than that of normal cell. It was suggested vanadate could be used as a potential radiosensitizer for tumor cells.

• Journal of Radiation Protection and Research
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• 제14권1호
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• pp.1-7
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• 1989

우라늄 피폭으로 발생하는 다뇨증과 급성 신부전증의 원인을 밝히기 위하여, 질산 우라늄을 정맥주사한 후 소변으로 배설되는 $Na^{+},\;K^{+}$의 전해질 양과 $Na^{+}-K^{+}$ adenosine triphosphatase($Na^{+}-K^{+}$ATPase) 활성도 변화를 측정하였다. 질산 우라늄 투여 24시간 이내에 $Na^{+},\;K^{+}$의 배설량이 크게 증가 하였고, 투여 3일 후에는 대조군과 비교하여 유의하게 감소하였다. 이때 $Na^{+},\;K^{+}$의 소변내 농도도 정상 대조군 범위 이하였다. 한편 $Na^{+}-K^{+}$ATPase활성은 투여 3일후에 고농도 질산 우라늄 투여 (30mg/kg BW) 시에만 감소 하였고, 저농도 투여군(5mg/kg BW, 15mg/kg BW) 에서는 활성 변화가 없었다.