• 제목/요약/키워드: fluorescence parameters

검색결과 251건 처리시간 0.029초

반복자기자극이 지방유래 중간엽 줄기세포 증식과 활성에 미치는 영향 (Effect of Repetitive Magnetic Stimulation on Proliferation and Viability of Adipose Tissue-Derived Stromal Cells)

  • 김수정;박해운;조윤우;이준하;서정민;신현진;강재훈;안상호
    • The Journal of Korean Physical Therapy
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    • 제21권3호
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    • pp.87-93
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    • 2009
  • Purpose: TThis study examined the effect of repetitive magnetic stimulation (RMS) on the viability and proliferative response of human adipose tissue-derived stromal cells (hATSCs) in vitro. Methods: The hATSCs were cultured primarily from human adipose tissue harvested by liposuction and incubated in a $37^{\circ}C$ plastic chamber. The cells were exposed to a repetitive magnetic field using a customized magnetic stimulator (Biocon-5000, Mcube Technology). The RMS parameters were set as follows: repetition rate=10Hz, 25Hz (stimulus intensity 100%= 0.1 Tesla, at 4cm from the coil), stimulated time= 1, 5, and 20 minutes. Twenty four hours after one application of RMS, the hATSCs were compared with the sham stimulation, which were kept under the same conditions without the application of RMS. The cells were observed by optical microscopy to determine the morphology and assessed by trypan blue staining for cell proliferation. The apoptosis and viability of the hATSCs were also analyzed by fluorescence-activated cell sorting (FACS) analysis of Annexin V and MTT assay. Results: After RMS, the morphology of the hATSCs was not changed and the apoptosis of hATSCs were not increased compared to the sham stimulation. The viability of the cells was similar to the cells given the sham stimulation. Interestingly, the level of hATSC proliferation was significantly higher in all RMS groups. Conclusion: The application of RMS may not cause a change in morphology and viability of hATSCs but can increase the level of cell proliferation in vitro. RMS might be useful as an adjuvant tool in combination with stem cell therapy without adverse effects.

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광환경조절에 따른 멸종위기식물 섬시호의 생리적 반응 (Physiological Responses of Bupleurum latissimum Nakai, Endangered Plants to Changes in Light Environment)

  • 이경철;왕명현;송재모
    • 생물환경조절학회지
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    • 제22권2호
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    • pp.154-161
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    • 2013
  • 차광처리에 의한 광환경 변화가 섬시호의 생리적 반응에 미치는 영향을 조사하기 위해, 차광막을 이용하여 무차광처리구(0%), 50%, 70% 및 90% 차광처리구를 설치하고 광합성 관련 인자, 수분이용효율, 엽육세포내 $CO_2$ 농도, 엽록소 형광반응 등을 조사하였다. 차광수준이 높아짐에 따라 낮은 광도에서 광합성을 수행하기 위해 암호흡의 저하로 인한 광보상점이 감소가 나타났으며, 광합성의 효율을 높이기 위해 순양자 수율과 엽록소 함량의 증가가 나타났다. 무차광처리구에서는 강한 광에 의한 수분손실을 막기 위해 기공전도도와 기공증산속도가 감소되었고, 엽육내의 $CO_2$를 효율적으로 광합성에 활용하지 못하는 것으로 나타났다. 또한 최대 광합성속도가 감소하는 광저해현상이 나타났으며, 여기 에너지의 전자전달이 원활하지 못하여 광으로 인한 피해가 예상된다. 90% 차광처리구 역시 점진적으로 광합성속도가 감소하여 7월에 가장 낮은 최대광합성속도를 보였는데, 이는 광선요구도보다 매우 적은 광 환경에서 계속 생장함으로서 광합성 능력이 저하되는 것으로 생각할 수 있으며, 50% 차광처리구의 경우 광합성에 효율적인 광환경이 제공되어 기공전도도와 기공증산속도가 증가하였고, 광화학반응 효율이 증가하는 등 광합성 활성이 높아진 것을 알 수 있어 생육에 보다 유리한 광 조건임을 알 수 있었다.

수용성 폴리파라시클로판류와 약물과의 상호작용(제 3보)-수용액 중 수용성 폴리파라시클로판류와 형광 소수 나프탈렌 유도체류와의 복합체 형성- (Interactions between Water-Soluble Polyparacyclophanes and Drugs (III) -Complex Formation of Water-Soluble Polyparacyclophanes with Fluorescent Hydrophobic Naphthalene Derivatives in Aqueous Solution-)

  • 전인구;이민화
    • Journal of Pharmaceutical Investigation
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    • 제19권2호
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    • pp.71-79
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    • 1989
  • Complex formation of water-soluble polyparacyclophanes bearing two diphenylmethane or two diphenyl ether skeletons with l-anilinonaphthalene-8-sulfonate (ANS) and 2-p-toluidinylnaphthalene-6-sulfonate (TNS) was investigated quantitatively to develop useful host compounds comparing with ${\alpha}\;-\;and\;{\beta}-cyc1odextrins$$({\alpha}-\;and\;{\beta}-CyDs$) in aqueous solution. Benesi-Hildebrand type analysis of the fluorescent intensity showed that the dissociation constants (Kd) of paracyclophane-ANS complexes were $1.55\;{\times}\;10^{-4}M$ for 1,6,20,25-tetraaza[6.1.6.1]paracyclophane(CPM 44) and $1.23\;{\times}\;10^{-4}M$ for 1,7,21,27-tetraaza[7.1.7.1]paracyclophane (CPM 55), and those of paracyclophane-TNS complexes were $6.99\;{\times}\;10^{-6}M$ for CPM 44 and $6.23\;{\times}\;10^{-5}M$ for CPM 55, in 1:1 molar ratio. On the other hand, the Kd values of 1,7,21,27-tetraaza-14,34-dioxa[7.1.7.1]paracyclophane (CPE 55)-ANS, 1,8,22,29-tetraaza-15,36-dioxa[8.1.8.1]paracyclophane (CPE 66)-ANS, CPE 55-TNS, CPE 66-TNS complexes were $1.75\;{\times}\;10^{-3}M$, $3.07\;{\times}\;10^{-3}M$, $3.75\;{\times}\;10^{-3}M$ and $2.15\;{\times}\;10^{-3}M$, respectively. On the contrary, the Kd values of ${\alpha}-CyD-ANS$, ${\beta}-CyD-ANS$, ${\alpha}-CyD-TNS$ and ${\beta}-CyD-TNS$ complexes were found to be $3.98\;{\times}\;10^{-2}M$, $1.05\;{\times}\;10^{-2}M$, $1.38\;{\times}\;10^{-2}M$ and $3.52\;{\times}\;10^{-4}M$, respectively. These results mean that the complexation of CPMs with ANS or TNS is by 5.6-1,975 fold stronger than that for ${\alpha}-or\;{\beta}-CyDs$, and the complex formation of CPEs with ANS or TNS is nearly same as or somewhat stronger than that for ${\alpha}-or\;{\beta}-CyDs$. From the Kd values determined at different temperatures, thermodynamic parameters were calculated and the complexation was found to be a spontaneous exothermic reaction. The effects of pH on Kd values of CPM 44-ANS, and CPM 55-ANS complexes were negligible in the range of pH 1.2-1.8. However, the Kd values of these complexes increased significantly with increasing ionic strength.

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Protective Effect of Propofol against Hypoxia-reoxygenation Injury in HaCaT Human Keratinocytes

  • Kim, Yong-Ho;Kang, Jin-Mo;Kim, In-Ryoung;Lee, Bo-Young;Yoon, Ji-Young;Kim, Cheul-Hong;Park, Bong-Soo
    • International Journal of Oral Biology
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    • 제39권2호
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    • pp.97-105
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    • 2014
  • The aim of this study was to determine the beneficial effect of propofol on human keratinocytes that have undergone hypoxia reoxygenation (H/R) injury and to investigate whether autophagy is associated with the protective mechanism. Thus, we evaluated how propofol influences the intracellular autophagy and apoptosis during the H/R process in the HaCaT cells. The cultured human keratinocyte cells were exposed to 24 h of hypoxia (5% $CO_2$, 1% $O_2$, 94% $N_2$) followed by 12 h of reoxygenation (5% $CO_2$, 21% $O_2$, 74% $N_2$). The experiment was divided into 4 groups: (1) Control=Normoxia ; (2) H/R=Hypoxia Reoxygenation ; (3) PPC+H/R=Propofol Preconditioning+Hypoxia Reoxygenation; (4) 3-MA+PPC+ H/R=3-MA-Methyladenine+Propofol Preconditioning+ Hypoxia Reoxygenation. In addition, Western blot analysis was performed to identify the expression of apoptotic pathway parameters, including Bcl-2, Bax, and caspase 3 involved in mitochondrial-dependent pathway. Autophagy was determined by fluorescence microscopy, MDC staining, AO staining, and western blot. The H/R produced dramatic injuries in keratinocyte cells. In our study, the viability of Propofol in H/R induced HaCaT cells was first studied by MTT assay. The treatment with 25, 50, and $100{\mu}M$ Propofol in H/R induced HaCaT cells enhanced cell viability in a dose-dependent manner and $100{\mu}M$ was the most effective dose. The Atg5, Becline-1, LC3-II, and p62 were elevated in PPC group cells, but H/R-induced group showed significant reduction in HaCaT cells. The Atg5 were increased when autophagy was induced by Propofol, and they were decreased when autophagy was suppressed by 3-MA. These data provided evidence that propofol preconditioning induced autophagy and reduced apoptotic cell death in an H/R model of HaCaT cells, which was in agreement with autophagy playing a very important role in cell protection.

디티오카르바메이트 유도체의 새로운 1,10-페난트롤린 팔라디움(II) 착물의 DNA 결합 성질 및 세포독성에 관한 연구 (DNA Binding Studies and Cytotoxicity of the Novel 1,10-phenanthroline Palladium(II) Complexes of Dithiocarbamate Derivatives)

  • Mansouri-Torshizi, Hassan;Saeidifar, Maryam;Ghasemi, Zahra Yekke;Khastan, Mahmood;Divsalar, Adeleh;Saboury, Ali Akbar
    • 대한화학회지
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    • 제55권1호
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    • pp.70-80
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    • 2011
  • 두 종류의 새로운 팔라디움(II) 착물인 [Pd(phen)(pip-dtc)]$NO_3$와 [Pd(phen)(mor-dtc)]$NO_3$ (여기서 phen은 1,10-페난트롤린, pip-dtc은 피페리딘디티오카르바메이트 음이온, mor-dtc은 모르폴린디티오카르바메이트 음이온을 가리킨다)를 합성하고 원소분석, 분광법(FT-IR, $^1H$ NMR, UV-Vis) 및 전기전도도 측정을 이용하여 특성을 조사하였다. 이 착물들에서, 디티오카르베이트 리간드는 팔라디움 (II) 중심과 두 개의 황 원자와 이중으로 배위 결합을 하고 있다. 이 착물의 세포독성을 K562 세포주를 이용하여 조사하였을 때 $IC_{50}$ 값이 시스플라틴보다 작았기 때문에 착물들의 송아지 흉선 DNA(CT-DNA)와의 결합 모드를 UV 흡광도 차이와 형광 분광법으로 조사하였다. 착물들은 DNA를 변형시키고, 협동결합을 하고, DNA에 끼어 들어간다. 또한 몇 가지 결합 및 열역학적인 변수들이 기술되었다.

실험견에서 Metoprolol 약리효과의 약동/력학적 검토 (Pharmacokinetic/Pharmacodynamic Analysis of Metoprolol in Dogs)

  • 오동진;장인진;이경훈;임동석;김형기;신상구;박찬웅;신재국
    • 대한약리학회지
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    • 제31권2호
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    • pp.251-259
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    • 1995
  • Pharmacokinetics and pharmacodynamics of metoprolol, a selective beta-l blocker, were examined for 360 minutes after intravenous bolus administration of metoprolol to 6 dogs. Plasma concentration and excreted amount in the urine metoprolol were measured by liquid chromatography with fluorescence detection. PR interval and heart rate were measured by ECG monitoring. Blood pressure was monitored through intraarterial catheter in femoral artery and cardiac output by thermodilution method using Swan-Ganz catheter. To analyze the effect site concentration-response relationship, plasma concentration and pharmacological effects were simultaneously fitted to a two pharmacokinetic compartment linked to pharmacodynamic model with NONLIN program. Results are as follows. 1) The plasma concentration of metoprolol after intrvenous injection decreased biexponentially. The terminal half-life estimated was $1.33{\pm}0.40$ hours and the volume of distribution at steady state (Vdss) and the total body clearance were $1.04{\pm}0.4\;L/kg,\;6.55{\pm}2.21\;L/hr$, respectively. The central compartment volume of distribution and peripheral compartment volume of distribution were $0.35{\pm}0.14L/kg\;and\;0.69{\pm}0.34L/kg$. The renal clearance and intercompartment clearance were $0.53{\pm}0.25\;L/min\;and\;0.35{\pm}0.19\;L/min$. 2) Simulated biophase concentration-response curve shows hyperbolic relationship and the estimated concentration-effect relationship was best explained by Emax model when the prolongation of PR interval and the reduction of the heart rate were used as pharmacodynamic parameters. Emax and EC50 were estimated to be $26.3{\pm}4.7\;msec\;and\;88.8{\pm}82.3\;g/ml$ for PR interval, and $48.7{\pm}18.8\;beats/min\;and\;113.5{\pm}78.7\;ng/ml$ for heart rate, respectively. 3) The changes of cardiac output-effect site concentration relationship was best fitted by a linear model and the slope of the relationship was $0.005{\pm}0.003$. Diastolic blood pressure-effect site concentration relationship was also explained by the linear model and the slope of the relationship was $0.038{\pm}0.034$.

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Bioequivalence and Pharmacokinetics of 70 mg Alendronate Sodium Tablets by Measuring Alendronate in Plasma

  • Yun Min-Hyuk;Woo Jong-Su;Kwon Kwang-Il
    • Archives of Pharmacal Research
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    • 제29권4호
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    • pp.328-332
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    • 2006
  • The bioequivalence and pharmacokinetics of alendronate sodium tablets were examined by determining the plasma concentration of alendronate. Two groups, consisting of 24 healthy volunteers, each received a 70 mg reference alendronate sodium tablet and a test tablet in a $2{\times}2$ crossover study. There was a 6-day washout period between doses. The plasma alendronate concentration was monitored for 7 h after the dose, using HPLC-Fluorescence Detector (FD). The area under the plasma concentration-time curve from time 0 to the last sampling time at 7 h $(AUC_{0-7h})$ was calculated using the linear-log trapezoidal rule. The maximum plasma drug concentration $(C_{max})$ and the time to reach $C_{max}(T_{max})$ were derived from the plasma concentration-time data. Analysis of variance was performed using logarithmically transformed $AUC_{0-7h}\;and\;C_{max}$, and untransformed $T_{max}$. For the test medication versus the reference medication, the $AUC_{0-7h}\;were\;87.63{\pm}29.27\;vs.\;102.44{\pm}69.96ng\;h\;mL^{-1}$ and the $C_{max}$ values were $34.29{\pm}13.77\;vs.\;38.47{\pm}24.39ng\;mL^{-1}$ respectively. The $90\%$ confidence intervals of the mean differences of the logarithmic transformed $AUC_{0-7h}$ and $C_{max}$ values were log 0.8234-log 1.1597 and log 0.8222-log 1.1409, respectively, satisfying the bioequivalence criteria guidelines of both the US Food and Drug Administration and the Korea Food and Drug Administration. The other pharmacokinetic parameters for the test drug versus reference drug, respectively, were: $t_{1/2},\;1.87{\pm}0.62\;vs.\;1.77{\pm}0.54\;h;\;V/F,\;2061.30{\pm}986.49\;vs.\;2576.45{\pm}1826.05\;L;\;CL/F,\;835.32{\pm}357.35\;vs.\;889.48{\pm}485.87\;L\;h^{-1}; K_{el},\;0.42{\pm}0.14\;vs.\;0.40{\pm}0.18\;h^{-1};\;Ka,\;4.46{\pm}3.63\;vs.\;3.80{\pm}3.64\;h^{-1};\;and\;T_{lag},\;0.19{\pm}0.09\;vs.\;0.18{\pm}0.06\;h$. These results indicated that two alendronate formulations(70-mg alendronate sodium) were biologically equivalent and can be prescribed interchangeably.

세포 배양 가능한 커버슬립형 초음파 변환자 (Cell-cultivable ultrasonic transducer integrated on glass-coverslip)

  • 이근형;박진형
    • 한국음향학회지
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    • 제42권5호
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    • pp.412-421
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    • 2023
  • 초음파 뇌 자극술을 통하여 뇌 심부의 국소 지역에 있는 뇌 세포의 활성화를 유도할 수 있으며, 이를 통하여 저하된 뇌 기능을 치료하는데 효과가 있음이 보고되어 왔다. 반면, 초음파 자극의 종류에 따라 신경 변조의 효율과 방향이 달라질 수 있음이 알려져 있어, 적절한 초음파 자극의 종류를 확립하는 연구가 중요하다. 따라서, 본 논문에서는 이를 효과적으로 최적화 하기 위해 세포 배양시 사용되는 커버슬립 기반의 초음파 변환자를 제안하고자 한다. 균일한 초음파 자극을 전도하기 위해서 폴리머 압전소자(Poly-vinylidene fluoride-trifluorethylene, PVDF-TrFE)를 스핀 코팅하고 패를린 절연층을 상단에 적층시켜 음압 출력을 극대화 시켰다. 개발된 초음파 변환자 융합 커버슬립은 초음파자극기 표면에 배양된 수십개의 신경세포에 균일하고 정확한 초음파 자극을 전달 할 수 있고, 자극에 따른 세포의 반응을 형광 현미경으로 실시간 관찰 가능하다. 따라서, 동일한 초음파 자극에 대한 세포의 반응 신호를 최대 수십개 세포로부터 동시에 획득 가능하므로, 반응 신호를 평균 한다면 낮은 강도의 초음파 자극에 따른 뇌 세포의 미세한 반응을 검출할 수 있을 뿐만 아니라, 초음파 변환자와 물의 표면 등에서 발생하는 정현파에 의한 자극의 왜곡 현상을 줄일 수 있어서 사용자가 원하는 초음파 자극을 정확하게 세포로 전달 가능하다. 이렇게 개발된 초음파 변환자를 통해 변환자 표면에 배양된 별세포에서 6 MHz, 0.2 MPa의 저강도 초음파 자극에 의해 유도된 칼슘 반응을 성공적으로 관찰할 수 있었다.

산화적 스트레스에 대한 생간건비탕가음양곽(生肝健脾湯加淫羊藿) (2:1)의 간보호효과 (SKT + EKE (2:1) protects oxidative stress induced-liver damage)

  • 박상미;정대화;진효정;김예림;김경순;황보민;김상찬
    • 대한한의학방제학회지
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    • 제32권1호
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    • pp.63-82
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    • 2024
  • Objective : Saengkankunbi-tang (SKT) is used as a traditional Korean herbal formula for treatment of liver diseases. We investigated the hepatoprotective effects of SKT plus Epimedium koreanum Nakai (EKE) against arachidonic acid (AA) + iron-mediated cytotoxicity in HepG2 cells and carbon tetrachloride (CCl4)-mediated acute liver damage in mice. Methods : The cyto-protective effects of SKT + EKE were determined by MTT assay, western blot and fluorescence activated cell sorting analysis. In mice, blood biochemistry and western blot were assessed in CCl4-induced acute hepatic damage. The animal groups included vehicle-treated control, CCl4, SKT (200 mg/kg/day), EKE (100 mg/kg/day), SKT (200 mg/kg/day) + EKE (100 mg/kg/day) and silymarin (200 mg/kg/day). Results : In HepG2 cells, pretreatment with SKT + EKE significantly suppressed cytotoxicity induced by AA + iron and reduced the expression of proteins related to apoptosis. In addition, pretreatment with SKT + EKE significantly prevented the increase in H2O2 production, GSH depletion, and lower mitochondrial membrane potential induced by AA + iron. In CCl4-induced liver damage mice, the administration of SKT + EKE prevented the liver damage by inhibition of hepatocyte damage and expression of apoptosis proteins in liver. More importantly, in vitro and in vivo assay, SKT + EKE showed significant effect compare with SKT alone or EKE alone in all parameters. Conclusions : These results indicated that SKT + EKE could protect against oxidative stress-induced liver damage, and SKT + EKE is more effective than SKT alone or EKE alone.

Expression of CsRCI2s by NaCl stress reduces water and sodium ion permeation through CsPIP2;1 in Camelina sativa L.

  • Kim, Hyun-Sung;Lim, Hyun-Gyu;Ahn, Sung-Ju
    • 한국작물학회:학술대회논문집
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    • 한국작물학회 2017년도 9th Asian Crop Science Association conference
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    • pp.194-194
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    • 2017
  • Camelina (Camelina sativa L.) is a potential bio-energy crop that has short life cycle about 90 days and contains high amount of unsaturated fatty acid which is adequate to bio-diesel production. Enhancing environmental stress tolerance is a main issue to increase not only crop productivity but also big mass production. CsRCI2s (Rare Cold Inducible 2) are cold and salt stress related protein that localized at plasma membrane (PM) and assume to be membrane potential regulation factor. These proteins can be divide into C-terminal tail (CsRCI2D/E/F/G) or no-tail group (CsRCI2A/B/C/H). However, function of CsRCI2s are less understood. In this study, physiological responses and functional characterization of CsRCI2s of Camelina under salt stress were analyzed. Full-length CsRCI2s (A/B/E/F) and CsPIP2;1 sequences were confirmed from Camelina genome browser. Physiological investigations were carried out using one- or four-week-old Camelina under NaCl stress with dose and time dependent manner. Transcriptional changes of CsRCI2A/B/E/F and CsPIP2;1 were determined using qRT-PCR in one-week-old Camelina seedlings treated with NaCl. Translational changes of CsRCI2E and CsPIP2;1 were confirmed with western-blot using the antibodies. Water transport activity and membrane potential measurement were observed by cRNA injected Xenopus laevis oocyte. As results, root growth rate and physiological parameters such as stomatal conductance, chlorophyll fluorescence, and electrolyte leakage showed significant inhibition in 100 and 150 mM NaCl. Transcriptional level of CsPIP2;1 did not changed but CsRCI2s were significantly increased by NaCl concentration, however, no-tail type CsRCI2A and CsRCI2B increased earlier than tail type CsRCI2E and CsRCI2F. Translational changes of CsPIP2;1 was constitutively maintained under NaCl stress. But, accumulation of CsRCI2E significantly increased by NaCl stress. CsPIP2;1 and CsRCI2A/B/E/F co-expressed Xenopus laevis oocyte showed decreased water transport activity as 61.84, 60.30, 62.91 and 76.51 % at CsRCI2A, CsRCI2B, CsRCI2E and CsRCI2F co-expression when compare with single expression of CsPIP2;1, respectively. Moreover, oocyte membrane potential was significantly hyperpolarized by co-expression of CsRCI2s. However, higher hyperpolarized level was observed in tail-type CsRCI2E and CsRCI2F than others, especially, CsRCI2E showed highest level. It means transport of $Na^+$ ion into cell is negatively regulated by expression of CsRCI2s, and, function of C-terminal tail is might be related with $Na^+$ ion influx. In conclusion, accumulation of NaCl-induced CsRCI2 proteins are related with $Na^+$ ion exclusion and prevent water loss by CsPIP2;1 under NaCl stress.

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