• 제목/요약/키워드: Intracellular chloride

검색결과 77건 처리시간 0.036초

사람 중성호성 백혈구의 기능에 있어서 Arachidonic Acid의 활성화 기전 (Activation Mechanism of Arachidonic Acid in Human Neutrophil Function)

  • 심재건;이정수;신용규;이광수
    • 대한약리학회지
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    • 제28권1호
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    • pp.91-102
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    • 1992
  • 칼슘을 함유하는 반응액에서 arachidonic acid는 뚜렷하게 superoxide와 $H_2O_2$의 생성을 자극하였고 NADPH oxidase를 활성화하였다. 칼슘이 없는 반응액에서 NADPH oxidase에 대한 arachidonic acid의 자극 작용은 나타나지 않았다. Arachidonic acid에 의하여 자극된 respiratory burst는 EGTA, TMB-8, verapamil, diltiazem, nifedipine, dibucaine, lidocaine, CCCP, 2,4-dinitrophenol, sodium arsenate, chlorpromazine, theophylline, $HgCl_2$, PCMB와 PCMBSA에 의하여 억제되었으나, tetrodotoxin, tetraethyl ammonium chloride와 procaine의 영향은 받지 않았다. EGTA는 거의 완전히 arachidonic acid에 의한 ${\beta}-glucuronidase$의 유리를 억제하였으며, verapamil, CCCP와 theophylline은 약간 억제하였으나, 이에 반하여 dibucaine은 유의한 효과를 나타내지 않았다. Arachidonic acid는 중성호성 백혈구로 부터 칼슘을 유리시켰으며, 이는 TMB-8에 의하여 감소되었다. Arachidonic acid에 의한 세포내 유리 칼슘 농도의 상승은 EGTA와 CCCP에 의하여 억제되었고 TMB-8에 의하여 약간 억제되었다. Arachidonic acid와 verapamil 또는 arachidonic acid와 dibucaine의 동시 첨가에 의하여 상승된 세포내 유리 칼슘 농도의 양은 arachidonic acid 단독에 의한 것보다 현저하였다. 이상의 결과로 부터 다양한 생화학적 변화가 arachidonic acid에 의하여 활성화된 중성호성 백혈구의 기능 표현에 관여할 것으로 시사된다. Arachidonic acid는 세포내 칼슘 저장고로부터 칼슘유리를 자극하여 세포내 유리 칼슘 농도를 상승시킬 것으로 추정된다. 중성호성 백혈구의 활성화증에 칼슘 유입과 유출은 동시에 이루어질 것으로 추정된다.

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질소고정균의 성장과 질소고정력에 대한 osmoprotectant의 영향 (Effects of Osmoprotectants on the Growth and Nitrogenase Activity of Rhizobium and Azospirillum under Osmotic Stress)

  • 갈상완;최영주
    • Applied Biological Chemistry
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    • 제41권1호
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    • pp.53-59
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    • 1998
  • 수종의 두과작물과 수도에서 분리한 Rhizobium 및 Azospirillum들의 내염성을 조사하고 salt stress에 의하여 유도되는 균주의 생리적 특성 및 식물이나 미생물에서 osmoprotectant 로 작용하는 proline, glycine betaine 및 glutamate가 질소고정균의 생육이나 질소고정력에 미치는 영향을 연구하였다. 분리된 대부분의 질소고정균들은 0.6 M NaCl 농도에서 생육이 현저히 감소되었지만 Acacia rhizobia sp86 은 1.4 M NaCl 농도에서도 생육이 가능하였다. Osmotic stress에 의하여 증가되는 intracellular 유리아미노산은 Rhizobium 및 Azospirillum에서 glutamate 였으며, 특히 Acacia rhizobia sp86은 salt stress에 의하여 5배정도 glutamate를 축적하였다. Osmoprotectant (proline, glycine betaine, glutamate)를 배지내에 1 mM 첨가함으로서 salt stress에 의하여 감소되는 질소고정균의 생육과 질소고정력을 방지하였으며, glycine betaine이 가장 효과적이었다.

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Calcium Signaling in Salivary Secretion

  • Kim, Jin Man;Lee, Sang-Woo;Park, Kyungpyo
    • Journal of Korean Dental Science
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    • 제10권2호
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    • pp.45-52
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    • 2017
  • Calcium has versatile roles in diverse physiological functions. Among these functions, intracellular $Ca^{2+}$ plays a key role during the secretion of salivary glands. In this review, we introduce the diverse cellular components involved in the saliva secretion and related dynamic intracellular $Ca^{2+}$ signals. Calcium acts as a critical second messenger for channel activation, protein translocation, and volume regulation, which are essential events for achieving the salivary secretion. In the secretory process, $Ca^{2+}$ activates $K^+$ and $Cl^-$ channels to transport water and electrolyte constituting whole saliva. We also focus on the $Ca^{2+}$ signals from intracellular stores with discussion about detailed molecular mechanism underlying the generation of characteristic $Ca^{2+}$ patterns. In particular, inositol triphosphate signal is a main trigger for inducing $Ca^{2+}$ signals required for the salivary gland functions. The biphasic response of inositol triphosphate receptor and $Ca^{2+}$ pumps generate a self-limiting pattern of $Ca^{2+}$ efflux, resulting in $Ca^{2+}$ oscillations. The regenerative $Ca^{2+}$ oscillations have been detected in salivary gland cells, but the exact mechanism and function of the signals need to be elucidated. In future, we expect that further investigations will be performed toward better understanding of the spatiotemporal role of $Ca^{2+}$ signals in regulating salivary secretion.

Biochemical Characterization of a Putative Calcium Influx Factor as a Diffusible Messenger in Jurkat Cells, Xenopis Oocytes, and Yeast

  • Kim, Hak-Yong
    • Animal cells and systems
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    • 제7권1호
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    • pp.75-79
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    • 2003
  • Highly purified high performance thin layer chromatography (HPTLC) fractions containing a putative calcium influx factor (CIF) were prepared from the Jurkat cells and Xenopus oocytes in which $Ca^{2+}$ stores were depleted by thapsigargin treatment and from the yeast in which intracellular $Ca^{2+}$ stores were also depleted by genetic means. Microinjection of the fractions has been shown to elicit $Ca^{2+}$ dependent currents in Xenopus oocytes. The nature of the membrane currents evoked by the putative CIF appeared to be carried by chloride ions since the current was blocked by the selective chloride channel blocker 1 mM niflumic acid and its reversal potential was about -24 mV. Injection of the calcium chelator 1,2-bis(2-aminophenoxy)ethane-N, N, N',N'-tetraacetic acid (BAPTA) eradicated the current activities, suggesting the current responses are entirely $Ca^2$-dependent. Moreover, the currents were sensitive to the removal of extracellular calcium, indicating the dependence on calcium entry through the plasma membrane calcium entry channels. CIF activities were insensitive to protease, heat, and acid treatments and to Dische-reaction whereas the activities were sensitive to nucleotide pyrophosphatase and hydrazynolysis. The fraction might have a sugar because it was sensitive to Molisch test and Seliwaniff's resorcinol reaction. From the above results, CIF as a small and stable molecule seems to have pyrimidine, pyrophosphate, and a sugar moiety.oiety.

농어(Lateolabrax japonicus) 아가미의 형태와 미세구조 (Morphology and Ultrastructure of Gill for Lateolabrax japonicus)

  • 강충배;김진구;김재원
    • Applied Microscopy
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    • 제39권4호
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    • pp.319-324
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    • 2009
  • 농어 아가미는 새파, 새궁, 새엽 및 새판으로 구성되어 있었다. 위쪽 새파수는 7~10개이며, 아래쪽 새파수는 13~18개로 조사되었다. 농어 아가미의 주요부위인 새엽과 새판(이차새엽)을 광학현미경과 투과전자현미경에서 관찰한 결과는 다음과 같다. 농어 아가미는 새궁(gill arch) 외연의 앞 뒤 2열로 된 긴 점막성의 빗살과 같은 모양으로 줄지어 있는 많은 수의 새엽(gill filament)을 가지고 있으며, 각 새엽은 전후 두개의 새판(gill lamellae)이 근접하여 두 열로 배열되어 있었다. 농어의 새판은 pavement cell, 기둥세포(pillar cell), 혈구세포, 점액세포(mucose cell) 및 염류세포(chloride cell) 등으로 구분되었다. 새판의 표면은 단층으로 큰 핵을 가진 pavement 세포로 구성되며, 상피표면의 glycocalyx로 덮혀진 미세융기를 가진다. 그리고 새판은 기둥세포와 혈구세포와 일정한 간격을 두고 분포하고 있었다. 점액세포는 새판에 드문드문 나타났지만 새엽에 많이 분포하고 있으며, 염류세포는 미토콘드리아와 tubular system이 잘 발달되어 있었다.

The role of $Na^+-Ca^{2+}$ exchange on calcium activated chloride current in single isolated cardiac myocyte in pulmonary vein of rabbit.

  • Kim, Won-Tae;Lee, Yoon-Jin;Ha, Jeong-Mi;Han Choe;Jang, Yeon-Jin;Park, Chun-Sik;Lee, Chae-Hun m
    • 한국생물물리학회:학술대회논문집
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    • 한국생물물리학회 2003년도 정기총회 및 학술발표회
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    • pp.37-37
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    • 2003
  • We have shown the $Ca^{2+}$-activated chloride current is present in cardiac myocyte in rabbit pulmonary vein (Kim et al., 2002). This current amplitude was increased as [N $a^{+}$]$_{i}$ was increased and we suggested this chloride current may be involve in the spontaneous action potential frequency change. Since this current is activated by the increase of intracellular $Ca^{2+}$, we would like to test what is the inducer of the increase of [C $a^{2+}$]$_{i}$ between a L-type $Ca^{2+}$-current or a reverse mode of N $a^{+}$-C $a^{2+}$ exchange current. White rabbit (1.5 kg) was used and anesthetized with Ketamin (100 mg/kg). Pulmonary vein (PV) was isolated and sleeve area between left atrium and PV was dissected. Using collagenase (Worthington 0.7 mg/cc), single cardiac myocytes were isolated. In the presence of 15 mM of N $a^{+}$, three steps of voltage pulses were applied (holding potential : -40 ㎷, -80 ㎷ for 50 msec, 30 ㎷ for 5 msec, 10 ㎷ steps from -70 ㎷ to 60 ㎷). The inward and outward tail current was activated after brief 5 msec prepulse. The outward tail current was blocked by the removal of extracellular chloride substituted by glucuronic acid or by a chloride channel blocker, 5 mM 9-AC. But the inward tail current was still remained even though the amplitude was decreased. The reversal potentials were changed to the direction of the change of chloride equilibrium potential ( $E_{Cl}$ ) but the shift of equilibrium potential was not enough to match to the theoretical equilibrium potential shift. In the presence of L-type $Ca^{2+}$ channel blocker, nifedipine 1 uM, inward tail currents were greatly reduced but the outward current tail currents were still remained. In the presence of N $a^{+}$-C $a^{2+}$ exchange current blocker, 10 uM KB-R7943, the inward and outward tail currents were blocked almost completely. We tried to test the $Ca^{2+}$sensitivity of the chloride current with various [C $a^{2+}$]$_{i}$ in pipette solution from 100 nM to 1 uM but we failed to activate $Ca^{2+}$-activated chloride currents even though the cell became contracted in the presence of 1 uM $Ca^{2+}$. From these results, we could conclude that the increase of [C $a^{2+}$]$_{i}$ to activate the outward $Ca^{2+}$-activated chloride current was mainly induced by the activation of the reverse mode of N $a^{+}$-C $a^{2+}$ exchanger, But for the increase of [C $a^{2+}$]$_{i}$ to activate the inward tail current, L-type $Ca^{2+}$ current may be the major provoking current. Since the cytosolic increase of [C $a^{2+}$]$_{i}$ through pipette solution have failed to activate $Ca^{2+}$-activated chloride current, this chloride current may have very low $Ca^{2+}$ sensitivity or a comparmental increase $Ca^{2+}$ such as in subsarcolemmal space may activate the chloride current. Since there are several reports and models that the increase of $Ca^{2+}$ in subsarcolemmal space would be over several to tens of uM, both possibility may be valid together.uM, both possibility may be valid together.

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TNF-$\alpha$에 의해 유도된 세포부착분자의 발현에 대한 Delphinidin chloride의 억제 효과 (Delphinidin Chloride Effects on the Expression of TNF-$\alpha$ Induced Cell Adhesion Molecules)

  • 고은경;채수철;서은선;나명석;이종빈
    • 환경생물
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    • 제27권1호
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    • pp.88-94
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    • 2009
  • 안토시아닌(Anthocyanin)은 플라보노이드계 화합물의 한 부류로 항산화, 항암 및 항궤양, 항당뇨, 중금속해독, 시력보호, 콜레스테를 저하 등의 다양한 생리활성을 가지는 것으로 보고되어 있다. 죽상경화과정은 염증성 사이토카인의 분비 또는 혈관손상으로 인한 백혈구의 부착과 이동을 통해 시작된다. 본 연구는 이러한 죽상경화의 초기과정에서 안토시아닌 혼합물 중 single compound인 delphinidin chloride (DC) 인간혈관 내피세포주(HUVEC, human umbilical vein endothelial cell line)에서 백혈구 부착과 관련이 있는 ICAM-1 (Intraceliular Adhesion Molecule-1)과 VCAM-1 (Vascular Adhesion Molecule-1) 발현에 미치는 영향에 대해 조사하였다. 세포독성이 없는 농도에서 TNF-$\alpha$에 의해 유도된 혈관 내피세포에 대한 단핵구의 부착정도를 측정하기 위해 monocyte-endothelial cell adhesion assay와 광학현미경을 이용한 형태학적 관찰을 한 결과 DC가 처리농도 의존적으로 부착을 억제하였다. 내피세포로부터 TNF-$\alpha$에 의해 유도된 세포부착 분자인 VCAM-1과 ICAM-1의 발현에 대한 영향을 western blot analysis 및 RT-PCR방법으로 비교 분석한 결과 VCAM-1과 ICAM-1의 단백질과 mRNA수준에서의 발현이 농도 의존적으로 감소되었다. 이러한 결과들을 종합해 볼 때 안토시아닌 중에서 DC를 실험한 결과 DE는 TNF-$\alpha$에 의해 유도된 내피세포의 ICAM-1과 VCAM-1 발현 억제효과를 확인할 수 있었다.

Effect of zinc and calcium on the intracelularly uptake of cadimium and growth of escherichia coli

  • Hong, Hyo-Bong;Brown, Lewis R.;Kim, Jong-Kyu
    • Journal of Microbiology
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    • 제33권4호
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    • pp.302-306
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    • 1995
  • E. coli was tested for their ability to uptake cadmium intracellularly, and the effect of zinc and calcium on cadmium toxicity to E. coli was observed. In addition, the effect of zinc and calcium on the uptake of cadimium was also studied. This study showed that living E. coli cells took up more cadmium than the dead cells. E. coli in the log phase uptake cadimiumm more actively than E. coli in the stationary phase. These results suggested that there may be metabolic reactions or compounds which encourage the uptake of cadimium. This study also showed that cadimium was sequestered by cell components of which molecular weight is about 30,000. Adding of zinc and calcium chloride reduced cadmium toxicity in E. coli and encouraged intracellular uptake by E coli. However adding of heavy metal solutions helped the microorganisms to adsorb more cadmium. Extremely high or low concentrations of zinc, however, did not affect cell viability.

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화학적 저산소증이 유도하는 뇌신경세포 손상에 있어서 미성숙 진귤 과피 발효 추출물의 보호 효과 (Anti-apoptotic effect of fermented Citrus sunki peel extract on chemical hypoxia-induced neuronal injury)

  • 고운철;이선령
    • Journal of Nutrition and Health
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    • 제48권5호
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    • pp.451-456
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    • 2015
  • Purpose: Neuronal apoptotic events induced by aging and hypoxic/ischemic conditions is an important risk factor in neurodegenerative diseases such as ischemia stroke and Alzheimer's disease. The peel of Citrus sunki Hort. ex Tanaka has long been used as a traditional medicine, based on multiple biological activities including anti-oxidant, anti-inflammation, and anti-obesity. In the current study, we examined the actions of fermented C. sunki peel extract against cobalt chloride ($CoCl_2$)-mediated hypoxic death in human neuroblastoma SH-SY5Y cells. Methods: Cell viability was measured by trypan blue exclusion. Expression of apoptosis related proteins and release of cytochrome c were detected by western blot. Production of intracellular reactive oxygen species (ROS) and apoptotic morphology were examined using 2',7'-dichlorofluorescin diacetate (DCF-DA) and 4',6-diamidino-2-phenylindole (DAPI) staining. Results: Exposure to $CoCl_2$, a well-known mimetic agent of hypoxic/ischemic condition, resulted in neuronal cell death via caspase-3 dependent pathway. Extract of fermented C. sunki peel significantly rescued the $CoCl_2$-induced neuronal toxicity with the cell viability and appearance of apoptotic morphology. Cytoprotection with fermented C. sunki peel extract was associated with a decrease in activities of caspase-3 and cleavage of poly (ADP ribose) polymerase (PARP). In addition, increase in the intracellular ROS and release of cytochrome c from mitochondria to the cytosol were inhibited by treatment with extract of fermented C. sunki peel. Conclusion: Based on these data, fermented C. sunki peel extract might have a protective effect against $CoCl_2$-induced neuronal injury partly through generation of ROS and effectors involved in mitochondrial mediated apoptosis.

삼채 뿌리의 항산화 및 수명연장 효과 (Antioxidant and Longevity Properties of the Root of Allium hookeri in Caenorhabditis elegans)

  • 이은별;김준형;양재헌;김영수;전현일;기별희;이성현;김유석;한순천;김대근
    • 생약학회지
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    • 제46권3호
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    • pp.234-242
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    • 2015
  • Allium hookeri (Liliaceae), native to India is widely cultivated in Korea lately as a medicinal plant. This study was performed to evaluate the antioxidant and lifespan extending effects of the root of A. hookeri. Methanol extract from the root of A. hookeri was successively partitioned as methylene chloride, ethyl acetate, n-butanol and H2O soluble fractions. Antioxidant effects of the fractions were measured by measuring the radical scavenging effect on 1,1-diphenyl-2-picrylhydrazyl (DPPH), and superoxide quenching activities. Lifespan extending effects of the fractions were investigated using Caenorhabditis elegans model system. In addition, stress resistant, superoxide dismutase (SOD) and catalase activities, and intracellular reactive oxygen species (ROS) levels were also studied. Furthermore, we had investigated aging-related factors such as reproduction, food intake, growth and movement of C. elegans to find any sample-induced significant change. Our results revealed that ethyl acetate soluble fraction from the root of A. hookeri showed the most potent antioxidant and lifespan extending effects, and this fraction elevated the most potent SOD and catalase activities of worms, and reduce intracellular ROS accumulation.