• Applied Biological Chemistry
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• 제39권3호
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• pp.189-194
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• 1996

세포의 활성을 좌우하는 세포질내의 칼슘농도 조절기작을 이해하기 위하여, 전갈 Leiurus quinquestriatus hebraeus (Lqh) 에서 얻은 전갈독소의 세포내 칼슘농도 조절기능과 관련된 효소들의 활성변화에 미치는 효과를 조사하였다. 칼슘펌프와 칼슘채널 효소활성은 돼지의 호흡기 상피세포에서 분리된 마이크로솜에서 측정되었으며, 전갈 독소 Lqh는 온전한 마이크로솜 시료의 경우 총 ATPase의 활성을 약 32% 증가시켰고, Triton X-100나 $Ca^{2+}$ ionophore A23187을 처리한 마이크로솜 시료에서는 총 ATPase의 활성을 약 28% 증가시켰다. Lqh 독소에 의한 ATPase의 활성증가는 $Ca^{2+}-ATPase$의 특이적 저해제인 thapsigargin의 처리로 완전히 저해되었으며, 이것으로 전갈독소 Lqh가 마이크로솜에 위치한 $Ca^{2+}-ATPase$의 활성을 특이적으로 증가시킴을 확인할 수 있었다. 이러한 결과는 결국 Lqh 독소가 $Ca^{2+}-ATPase$의 활성화에 의해 마이크로솜의 $^{45}Ca^{2+}$ uptake를 증가시킬 것이라는 예상을 가능하게 하나, 실제로는 Lqh의 처리가 마이크로솜의 $^{45}Ca^{2+}$ uptake를 오히려 약 30% 감소시켰다. 예상되었던 Lqh에 의한 $^{45}Ca^{2+}$ uptake의 증가는 오직 $InsP_3$ 수용체 칼슘채널의 저해제인 Heparin의 존재시에만 얻어졌다. 이것은 Lqh 독소가 $Ca^{2+}-ATPase$의 활성화에 따른 $^{45}Ca^{2+}$ uptake를 증가시킴과 동시에 또한 $^{45}Ca^{2+}$ release를 유발시킴을 의미하며, Lqh 독소는 실제로 $InsP_3$에 의한 칼슘채널 활성화시에 얻어지는 것과 같은 정도의 $^{45}Ca^{2+}$ release를 유발시킴이 확인되었다. 위의 결과들로부터 Lqh독소에는 적어도 두 가지의 활성성분이 포함되어 있음을 알 수 있었고, 그중 하나는 $InsP_3$ 수용체 칼슘채널의 활성을, 다른 하나는 칼슘펌프의 활성을 촉진시킨다는 결론을 얻었다. 현재 Lqh 독소의 활성성분에 대한 화학적, 전기생리학적 특성들이 연구되어지고 있다.

• 한국식품위생안전성학회지
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• 제11권2호
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• pp.83-87
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• 1996

The objective of this study is to investigate the direct effects of green tea catechins(GTC) on vascular smooth muscle tension and 45Ca2+ Uptake in rat aorta. The methods used in this study are isometric tension measurements using physiograph, Lanthanum method for 45Ca2+(2 uCi/ml) uptake measurement in rat aorta. GTC modified tension induced by 40 mM KCl or 1 uM norepinephrine in rat aorta. Low concentrations of GTC(<0.5mg/ml) increased tension by 40 mM KCl or 1 uM norepinephrine, individually. However, high conecentration of GTC(>0.5 mg/ml) inhibiited tension by 40 mM KCl or 1 uM norepinephrine, individually. GTC increased 45Ca uptake induced by 40 mM KCl in a dose-dependent manner. From these results, GTC has the dual actions in vascular smooth muscle in vitro. Low concentrations of GTC augments tension by K or norepinephrine. However, high concentrations of GTC inhibits tension by K or norepinephrine GTC may have Ca2+ channel activation, action, which may result in unphysiological vasodilation by Ca2+ overload in vascular smooth muscle.

• Archives of Pharmacal Research
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• 제12권2호
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• pp.128-134
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• 1989

$Na^+-Ca^{2+}$ exchange process in sarcolemmal vesicles isolated from mesenteric arteries of Wistar-Kyoto normotensive(WKY) and spontaneously hypertensive rats(SHR) was investigated. The sarcolemmal fractions isolated after homogenization and sucrose density gradient centrifugation were enriched with 5'-nucleotidase and ouabain sensitive, $K^+-dependent$ phosphatase activities. When the vesicles were loaded with $Na^+$, a time dependent $Ca^{2+}$ uptake was observed. However, very little $Ca^{2+}$ uptake was observed when the vesicles were loaded with $K^+$, or $Ca^{2+}$ uptake of the $Na^+-loaded$ vesicles was carried out in high sodium medium so that there was no sodium gradient. When the vesicles loaded with $Ca^{2+}$ by $Na^+-Ca^{2+}$ exchange were diluted into potassium medium containing EGTA, $Ca^{2+}$ was rapidly released from the vesicles. $Na^+-dependent\;Ca^{2+}$ uptake was increased in SHR compared to WKY, but passive efflux of preaccumulated $Ca^{2+}$ from the vesicles was decreased in SHR. The data indicate that the membrane vesicles of rat mesenteric arteries exhibit $Na^+-Ca^{2+}$ exchange activity. It is also suggested that changes of this process in vascular smooth muscle cell membrane of SHR may be involved in higher intracellular $Ca^{2+}$ concentration and higher basal tone in SHR.

• 약학회지
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• 제28권3호
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• pp.129-138
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• 1984

Specific [$^{3}H$] ouabain binding and $Ca^{2+}$ -uptake were measured to elucidate the role of high affinity [$^{3}H$] ouabain binding site in rat cardiac myocytes which contain 65% of rod cells. High affinity [$^{3}$H] ouabain binding site, which is about 3% of total pump sites, with apparent dissociation constant ($K_{D}$) of $1.1{\times}10^{-7}M$ and maximum binding site concentration (Bmax) of 1.2 pmol/mg protein ($1.754{\times}10^{5}cells$) were identified. At the concentration of $10^{-7}M$ to $10^{-4}M$, ouabain produced concentration dependent increase in $Ca^{2+}$-uptake of myocytes. The effect of ouabain on $Ca^{2+}$-uptake was not effected by membrane depolarization (elevated K+ in incubation medium) or verapamil. These results suggest that in rat ventricular myocytes the ouabain receptor complex to high affinity site may increase Na+ - $Ca^{2+}$ exchange across the sarcolemmal membrane by inhibition of Na+, K+ - ATPase.

• 한국동물학회지
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• 제20권2호
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• pp.101-107
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• 1977

토끼의 골격근 小胞體의 ATPase 活性과 Ca 輸送에 대한 sodium azide, cAMP, G-strophanthin 및 dicumarol의 영향을 측정하였다. Sodium azide(0.05mM)와 G-strophanthin(0.25nM)은 ATPase活性과 Ca 輸送能에 아무런 영향도 미치지 아니하였다. cAMP$(1 \\times 10^-6 M \\sim 5 \\times 10^-4 M)$는 ATPase 活性에는 아무런 영향도 미치지 않았으나 Ca 輸送은 억제하였다. Dicumarol(0.05mM)도 ATPase 活性에는 영향이 없었으나 小胞體의 $8,000 \\sim 12,000 \\times G$分劃에서의 Ca 輸送을 억제하였다.

• Clinical and Experimental Reproductive Medicine
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• 제21권3호
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• pp.285-296
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• 1994

Follicular oocytes were released from the graafian follicles of ovaries from 3-4 weeks old mice. The spontaenous maturation of these follicular oocyes was inhibited by the treatment of dbcAMP and progesterone and these oocytes were cultured for 2-8hr in the Modified Hank's balanced salt solution(MHBS). Ethylenediaminetetraaceticacid(EDTA) and calmoudulin antagonist, trifluoperazine (TFP) were treated to the culture medium in order to investigate whether these chemical agents inhibit calcium uptake into the oocyte and oocyte maturation. $^{45}Ca^{2+}$, 10-${\mu}$Ci/ml was added to the culture medium during the culture period. $^{45}Ca^{2+}$uptake into the oocytes was examined whether and when various kind of oocyte maturation inhibiting agents inhibit or stimulate the influx of calcium into oocytes. Dibutyryl cAMP and progesterone decrease $^{45}Ca^{2+}$uptake into the oocytes and synergistic inhibiting effect of dbcAMP and progesterone was prominent at much lower dosages. Calcium uptake into oocytes seems to be higher during first 2 hour culture period rather than next 4hr culture. After 8hr culture, calcium uptake level of the oocytes which GVBD already took place gradually approached to the level of those which were maintained at GV by the treatment of dbcAMP and progesterone. However, $^{45}Ca^{2+}$uptake into the GV maintained oocytes did not change at all even after 8hr culture period. In addition, calcium chelating agent, EDTA inhibited calcium uptake into oocytes as well as nuclear maturation of oocytes. Lower dosage used in the present study did not inhibit calcium uptake as well as oocyte maturation.

• The Korean Journal of Physiology and Pharmacology
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• 제6권5호
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• pp.261-267
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• 2002

The aim of the present study was to investigate the interaction of $estradiol-17{\beta}-bovine$ serum albumin $(E_2-BSA)$ and calcitropic hormones, such as parathyroid hormone, calcitonin, and vitamin D, in regulation of $Ca^{2+}$ uptake in primary cultured renal proximal tubule cells. Statistically significant increase in $Ca^{2+}$ uptake was found from 2 hours after $(E_2-BSA)\;(10^{-9}\;M)$ treatment, while $estradiol-17{\beta}\;(10^{-9}\;M)$ did not affect. Treatment of the cells with $(E_2-BSA)\;(10^{-9}\;M)$ together with parathyroid hormone (PTH) $(10^{-8}\;M),$ vitamin D $(10^{-8}\;M),$ or calcitonin $(10^{-8}\;M)$ significantly stimulated $Ca^{2+}$ uptake by 32.50%, 29.30%, or 27.75%, respectively, compared with the control. However, calcitropic hormones did not exhibit any synergistic effect on the E2-BSA-induced stimulation. $E_2-BSA$ significantly increased cAMP generation and PKC activity. The stimulatory effect of cotreatment of $E_2-BSA$ and PTH or vitamin D was blocked by SQ22536 (an adenylate cyclase inhibitor) and staurosporine (a PKC inhibitor), but the effect of cotreatment of $E_2-BSA$ and calcitonin was not blocked. Furthermore, 8-Br-cAMP and TPA (an artificial PKC promoter) increased $Ca^{2+}$ uptake by 25.51% and 16.47%, respectively, compared with the control. In conclusion, $E_2-BSA$ combined with calcitropic hormones regulated $Ca^{2+}$ uptake partially via cAMP and PKC-dependent mechanisms in renal proximal tubule cells.

• Clinical and Experimental Reproductive Medicine
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• 제18권2호
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• pp.153-162
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• 1991

The present study was undertaken to clarify the role of calcium ion as a factor for the maturation of follicle-enclosed mouse oocytes. Follicles were isolated with two sharp needles under a stereomicroscope from mouse(ICR) ovaries which were treated PMSG 5 IU 45 hours previously. Isolated follicles were cultured for 14-16 hours in an organ culture system at $37^{\circ}C$, 5% $CO_2$ in air and in a 100% humidified incubator by treatment of hCG, EDTA and $^{45}Ca^{++}$. Culture medium was Modified Hank's Balanced Salt Sol. (MHBS) and addition of hCG (human chorionic gonadotropin) was made into two doses level 0.4 IU and 0.8IU from the stock sol. and also $^{45}Ca^{++}$ was treated in the culture medium. To explain the role of calcium, calcium chelating agent EDTA was treated to the culture of the mouse follicle-enclosed oocytes. Two observations were made in the present study; nucleus phase and $^{45}Ca^{++}$ uptake into the oocyte. HCG induced oocyte maturation in the follicle about two folds as much as the control group, whereas there is no difference in oocyte maturation between 0.4 IU and 0.8 IU of hCG. Optimum level of hCG seems to be 0.4 IU/ml in the mouse follicle culture. HCG stimulated $^{45}Ca^{++}$ uptake into the oocyte of the follicles by two folds. $^{45}Ca^{++}$ uptake in the control group is about 2.5 folds in comparison of the EDTA(1.71mM) treated group. However, calcium uptake in the EDTA treated groups tends to increase depending on the decrease of EDTA concentration. These observations suggest that firstly, hCG stimulates maturation of the oocyte of the follicle, secondly, $Ca^{++}$ influx is induced by hCG and thirdly, $Ca^{++}$ influx by the treatment of EDTA decreases as a dosage-dependent process. This $Ca^{++}$ uptake may take place by the changes of permeability which was induced by hCG treatment. That is, $Ca^{++}$ influx may trigger the resumption of oocyte maturation. It is further necessary in the future study how this $Ca^{++}$ uptake is induced by hCG and increases permeability of the follicle and oocyte.

• Archives of Pharmacal Research
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• 제6권1호
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• pp.69-73
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• 1983

It was reported from our laboratory that the rate of deterioration of the force of contraction was slower in heart from Panax ginseng extract treated rats. Present investigation was designed to elucidate the mechanism of the slow deterioration of contractility of ginseng treated hearts. Therefore, $^{45}Ca^{2+}$ Uptake by sarcoplasmic reticulum (SR) isolated from ginseng treated rate and control rats was studied. Rate weighing 150-250g were administered orally with ginseng ethanol extract (100mg/kg) for 10 days. Cardiac SR was isolated by differential centrifugation and $^{45}Ca^{2+}$ uptake was assessed by the Millipore method. Freshly isolated SR from treated as well as control animals did not show any differences, but after incubation for 30 and 60 min at 37.deg.C, $^{45}Ca^{2+}$ uptake of control animal SR was found to be greatly depressed. The SR of treated animal possessed a greater degree of resistance to incubation. Thus it can be concluded that ginseng may have an ability to sustain the normal function of the heart by sustaining Ca accumulation by SR involved with the excitationcontraction coupling processes.

• 한국식품영양학회지
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• 제15권2호
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• pp.89-96
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• 2002

Ca-alginate bead로 소금의 흡착에 미치는 영향조건을 검토한 결과는 다음과 같다. Ca-alginate beads에 의한 소금 흡착은 시간이 경과함에 따라 증가하였으며, 10분 후4.0g으로 최고의 흡착량을 나타내었다. 0.2M CaCl$_2$, 0.2M BaCl$_2$, 0.2M FeCl$_3$및 0.2M MgCl$_2$등의 경화용액으로 조제한 bead에 의한 소금 흡착량은 Fe-alginate beads가 5.6g으로 제일 높았으나 bead가 쉽게 부서지는 단점이 있었고, MgCl$_2$용액으로는 bead가 만들어지지 않았다. 그리고 0.2M CaCl$_2$, 0.2M BaCl$_2$및 0.2M SrCl$_2$용액으로 만든 bead는 각각 4.2g 정도의 대등한 흡착량을 나타내었다. CaCl$_2$경화 용액이 0.1M, 0.2M 및 1M 일때 소금 흡착량은 각각 4.8g, 4.2g 및 4.1g으로 나타났다. Alginate의 농도를0.6%, 1% 그리고 2%로 하여 제조한 비드로 소금 흡착량은 2.8g, 4.0g, 4.4g으로 각각 나타났으며, 그리고 bead의 크기를 각각 2.5mm, 3.5mm 그리고 4.5mm로 제조하여 소금의 흡착량을 살펴본 결과 각 크기별 모두 4.0~4.2g로 차이가 없었다. 초기 소금의 농도 4%, 8%, 12%그리고 16%에서, 각각 소금의 흡착율은 30%, 28%, 27% 그리고 25%이었으며, pH에 따른 염분의 흡탁율은 산성(pH 4.0) 및 중성(pH 6.8) 영역에서 보다는 염기성(pH 10.0)에서 더 높았다. 된장으로부터 내염성 세균을 분리한 후 alginate로 고정화한 beads와 비고정화한 bead와의 염분 흡착량은 고정화한 bead에 의한 염분 초기흡착속도가 보다 낮았으며, Ca-alginate bead 제조시 경화용액에 머무는 시간이 길수록 염분 흡착율은 감소하는 것으로 나타났다. 또한 1회 사용한 bead를 증류수에 하루 동안 방치 후 이 bead를 재이용 함에 따라 염분 흡착량은 점점 감소하였다. 시료를 된장으로 하여 0시간, 3시간, 6시간, 12시간 및 24시간 후 소금 흡착율은 시간의 경과에 따라 더불어 증가하였다. 또한 소금이 감소된 된장의 pH를 측정한 결과, 4.90, 5.00, 5.01, 5.02, 그리고 5.03이였으며, 적정산도는 0.1N-NaOH 소모량이 4ml, 3.4ml, 3.2ml, 3.0ml 그리고 3.0 ml이었다. 저염 된장의 아미노태 질소를 적정 한 결과, 원료 된장이 840mg/된장 100g, 740mg/된장 100g, 630mg/된장 100g, 그리고 530mg/된장 100g으로 줄어들었다. 각 시료별 염분 흡착율은 된장이 다른 시료(Doengjang 100%, Kochujang 86%, Soy-sauce 78% and Jeotkal 71%) 보다 가장 높게 나타났다.