• Title/Summary/Keyword: Intracellular uptake

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Liposome/Tat Complex for Facilitating Genistein Uptake into B16 Melanoma Cells

  • Park, Young-Mi;Kang, Myung-Joo;Moon, Ki-Young;Park, Sang-Han;Kang, Mean-Hyung;Choi, Young-Wook
    • Journal of Pharmaceutical Investigation
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    • v.41 no.4
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    • pp.205-210
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    • 2011
  • Genistein (GT), a major isoflavone found in soybeans, has a potent antioxidant effect that protects the skin from UV-induced damages and malignant melanoma. In order to enhance the cellular uptake of GT, liposome/Tat complexes were prepared by an electrostatic interaction of anionic liposome (DMPC/DCP, 9:1 in molar ratio) with Tat peptide (0.02 to 0.08 mole), one of the well-known cell penetrating peptide (CPP). As the amount of Tat increased, the size increased but the zeta potential decreased. In vitro release study with dialysis membrane elicited GT release from liposomal preparations in a controlled manner. The addition of Tat increased GT release, especially for the initial period. In the cellular uptake study by incubating B16 melanoma cells with various liposomal preparations containing GT, B16 melanoma cells demonstrated a time-dependent increase of drug accumulation. Compared to the aqueous GT suspension, intracellular uptake was substantially enhanced by anionic liposomal formulation and further increased by the complex formulation. Therefore, liposome/ Tat complex might be a good candidate for facilitating intracellular drug delivery.

Effect of pH on the Vascular Tone and $^{45}Ca$ Uptake in the Aorta of Spontaneously Hypertensive Rats

  • Chang, Seok-Jong;Jeon, Byeong-Hwa;Kim, Se-Hoon;Kim, Hoe-Suk;Park, Hae-Kun
    • The Korean Journal of Physiology
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    • v.28 no.2
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    • pp.169-179
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    • 1994
  • The effect of extracellular and intracellular pH on vascular tone and $^{45}Ca$ uptake were investigated in aortic strips and dispersed single aortic smooth muscle cells of spontaneously hypertensive rats (SHR) and aged-matched Wistar-Kyoto rats (WKY). The contraction produced by a change of extracellular pH (pHo) in the range of $6.5{\sim}8.3$ was estimated by comparison with the level of vascular tone at pH 7.4. Contraction was induced below pHo 6.5 in WKY, pHo 7.1 in SHR, and over pHo 8.0 on both strains. The amplitude of contraction induced by high pHo (over pHo 7.7) was similar in SHR and WKY, but that induced by low pHo (below pHo 7.1) in SHR was greater than that in WKY. Either high pHo- or low pHo-induced contractions in WKY and SHR were not induced in the Ca-free Tyrode's solution and were induced by the addition of Ca. $^{45}Ca$ uptake increased progressively as pHo was increased from 6.8 to 8.1 in the single aortic smooth muscle cells of WKY and SHR. $NH_4Cl$ induced a gradually developing contraction in a dose-dependent manner $(5\;mM{\sim}30\;mM)$ and the removal of $NH_4Cl$ induced transient contraction was followed by profound relaxation in the aortic rings of both strains. The contractions induced by $NH_4Cl$ or by the removal of $NH_4Cl$ in SHR were significantly greater than that in WKY. These contractions were not induced in Ca-free Tyrode's solution. $^{45}Ca$ uptake was increased by $NH_4Cl$ (20 mM) and was not changed by the removal of $NH_4Cl$ (20 mM) in the aortic strips of WKY and SHR. As a summary of above results, the vascular tone of SHR was more sensitive to the change pHi and pHo than that of WKY. The contractions induced by change of extracellular or intracellular pH depended on extracellular Ca in the aorta of SHR nnd WKY. However, the Ca uptake was in accord with the changes of contraction but increase in contraction by low pH was not accompanied by an increase in Ca uptake in both strains.

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N,N-Dimethyl-D-ribo-phytosphingosine Modulates Cellular Functions of 1321N1 Astrocytes

  • Lee, Yun-Kyung;Kim, Hyo-Lim;Kim, Kye-Ok;Sacket, Santosh J.;Han, Mi-Jin;Jo, Ji-Yeong;Lim, Sung-Mee;Im, Dong-Soon
    • Biomolecules & Therapeutics
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    • v.15 no.2
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    • pp.73-77
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    • 2007
  • N,N-Dimethyl-D-ribo-phytosphingosine (DMPH) is an N-methyl derivative of sphingosine. In the present paper, we studied effects of DMPH on intracellular Ca$^{2+}$ concentration, pH, glutamate uptake, and cell viability in human 1321N1 astrocytes. DMPH increased intracellular Ca$^{2+}$ concentration and cytosolic pH significantly in a dose-dependent manner. DMPH also inhibited glutamate uptake by 1321N1 astrocytes. Finally, treatment of cells with DMPH for 24 h reduced viability of cells largely and concentration-dependently. In summary, DMPH increased intracellular Ca$^{2+}$ concentration and pH, inhibited glutamate uptake and evoked cytotoxicity in 1321N1 astrocytes. Our observations with DMPH in the 1321N1 astrocytes would enhance understanding of DMPH actions in the brain.

Magnesium Uptake by the Green Microalga Chlorella vulgaris in Batch Cultures

  • Ayed, Hela Ben Amor-Ben;Taidi, Behnam;Ayadi, Habib;Pareau, Dominique;Stambouli, Moncef
    • Journal of Microbiology and Biotechnology
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    • v.26 no.3
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    • pp.503-510
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    • 2016
  • The accumulation (internal and superficial distribution) of magnesium ions (Mg2+) by the green freshwater microalga Chlorella vulgaris (C. vulgaris) was investigated under autotrophic culture in a stirred photobioreactor. The concentrations of the three forms of Mg2+ (dissolved, extracellular, and intracellular) were determined with atomic absorption spectroscopy during the course of C. vulgaris growth. The proportions of adsorbed (extracellular) and absorbed (intracellular) Mg2+ were quantified. The concentration of the most important pigment in algal cells, chlorophyll a, increased over time in proportion to the increase in the biomass concentration, indicating a constant chlorophyll/biomass ratio during the linear growth phase. The mean-average rate of Mg2+ uptake by C. vulgaris grown in a culture medium starting with 16 mg/l of Mg2+ concentration was measured. A clear relationship between the biomass concentration and the proportion of the Mg2+ removal from the medium was observed. Of the total Mg2+ present in the culture medium, 18% was adsorbed on the cell wall and 51% was absorbed by the biomass by the end of the experiment (765 h). Overall, 69% of the initial Mg2+ were found to be removed from the medium. This study supported the kinetic model based on a reversible first-order reaction for Mg2+ bioaccumulation in C. vulgaris, which was consistent with the experimental data.

Toxicity and Biomedical Imaging of Fluorescence-Conjugated Nanoparticles in Hematopoietic Progenitor Cells

  • Min, Gye-Sik;Kim, Dong-Ku
    • Reproductive and Developmental Biology
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    • v.35 no.4
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    • pp.503-510
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    • 2011
  • Cellular uptake of nanoparticles for stem cell labeling and tracking is a critical technique for biomedical therapeutic applications. However, current techniques suffer from low intracellular labeling efficiency and cytotoxic effects, which has led to great interest in the development of a new labeling strategy. Using silica-coated nanoparticles conjugated with rhodamine B isothiocyanate (RITC) (SR), we tested the cellular uptake efficiency, biocompatibility, proliferation or differentiation ability with murine bone marrow derived hematopoietic stem/progenitor cells. The bone marrow hematopoietic cells showed efficient uptake with SR with dose or time dependent manner and also provided a higher uptake on hematopoietic stem/progenitor cells. Biocompatibility tests revealed that the SR had no deleterious effects on cell cytotoxicity, proliferation, or multi-differentiation capacities in vitro and in vivo. SR nanoparticles are advantageous over traditional labeling techniques as they possess a high level of cellular internalization without limiting the biofunctionality of the cells. Therefore, SR provides a useful alternative for gene or drug delivery into hematopoietic stem/progenitor cells for basic research and clinical applications.

Inhibitory Effect of Rubus Coreanus on Compound 48/80- or Anti-DNP IgE-Induced Mast Cell Activation (Compound 48/80과 anti-DNP IgE로 유도되는 비만세포 활성화에 대한 복분자의 억제효과)

  • Li, Guang Zhao;Chai, Ok Hee;Song, Chang Ho
    • IMMUNE NETWORK
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    • v.4 no.2
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    • pp.100-107
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    • 2004
  • Background: The fruit of Rubus coreanus (RC), a perennial herb, has been cultivated for a long time as a popular vegetable. The anti-allergy mechanism of RC is unknown. The purpose of this study is to investigate the inhibitory effect of RC on compound 48/80- or anti-DNP IgE-induced mast cell activation. Methods: For this, influences of RC on the compound 48/80-induced degranulation, histamine release, calcium influx and the change of the intracellular cAMP (cyclic adenosine-3',5' monophosphate) levels of rat peritoneal mast cells (RPMC) and on the anti-DNP IgE-induced histamine release of RPMC were observed. Results: The pretreatment of RC inhibited compound 48/80-induced degranulation, histamine release and intracelluar calcium uptake of RPMC. The anti-DNP IgE-induced histamine release of RPMC was significantly inhibited by pretreatment of RC. The RC increased the level of intracellular cAMP of RPMC, and the pretreatment of RC inhibited compound 48/80-induced decrement of intracellular cAMP of RPMC. Conclusion: These results suggest that RC contains some substances with an activity to inhibit the compound 48/80- or anti-DNP IgE-induced mast cell activitation. The inhibitory effects of RC are likely due to the stabilization of mast cells by blocking the calcium uptake and enhancing the level of intracellular cAMP.

Effects of Chronic Lead Exposure on Glutamate Release and Uptake in Cerebellar Cells of Rat Pups

  • Yi, Eun-Young;Lim, Dong-Koo
    • Archives of Pharmacal Research
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    • v.21 no.2
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    • pp.113-119
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    • 1998
  • Changes in the release and uptake of glutamate in cerebellar granule and glial cells of offspring of lead-exposed mothers were determined. In cultured cerebellar granule cells exposed to lead for 5 days, glutamate release was less influenced upon N-methyl-D-aspartate (NMDA) stimulation than that in the control. Although the NMDA-stimulated release of glutamate in cerebellar granule cells prepared from lead-exposed first generation pups was not different from that of the control group, the S-nitroso-N-acetylpenicillamine (SNAP)-stimulated release of glutamate in cerebellar granule cells obtained from lead-treated pups was less elevated than that in the control. Furthermore, in cerebellar granule cells obtained from lead-exposed second generations pups, glutamate release did not respond to both NMDA and SNAP stimulation. In cerebellar glial cells exposed to lead, the basal glutamate uptake was not changed. However, the L-trans-pyrollidine-2,4-dicarboxylic acid (PDC)-blocking effects was significantly reduced. In glial cells obtained from lead-exposed pups, the glutamate uptake was also less blocked by PDC than that in the control. Further decreases in PDC-blocking effects were observed in cerebellar glial cells obtained from lead-treated second generation pups compared to those from the control group. These results indicate that lead exposure induces the changes in the sensitivities of the glutamate release and uptake transporter. In addition, these results suggest that lead exposure might affect the intracellular signalling pathway and transmission in glutamatergic nervous system.

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Effects of Anions on PAH Transport in Rabbit Kidney Cortical Slices (가토 신피질 절편에서 PAH$(\rho-aminohippuric\;acid)$ 이동에 미치는 음이온의 영향)

  • Suh, Duk-Joon;Lee, Sang-Ho;Sung, Ho-Kyung
    • The Korean Journal of Physiology
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    • v.19 no.1
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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.

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