• The Korean Journal of Physiology and Pharmacology
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• 제2권4호
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• pp.529-539
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• 1998

The effects of membrane surface charge originated from lipid head groups on ion channels were tested by analyzing the activity of single large conductance $Ca^{2+}-activated\;K^+$ (maxi K) channel from rat skeletal muscle. The conductances and open-state probability ($P_o$) of single maxi K channels were compared in three types of planar lipid bilayers formed from a neutral phosphatidylethanolamine (PE) or two negatively-charged phospholipids, phosphatidylserine (PS) and phosphatidylinositol (PI). Under symmetrical KCl concentrations $(3{\sim}1,000\;mM)$, single channel conductances of maxi K channels in charged membranes were $1.1{\sim}1.7$ times larger than those in PE membranes, and the differences were more pronounced at the lower ionic strength. The average slope conductances at 100 mM KCl were $251{\pm}9.9$, $360{\pm}8.7$ and $356{\pm}12.4$ $(mean{\pm}SEM)$ pS in PE, PS and PI membranes respectively. The potentials at which $P_o$ was 1/2, appeared to have shifted left by 40 mV along voltage axis in the membranes formed with PS or PI. Such shift was consistently seen at pCa 5, 4.5, 4 and 3.5. Estimation of the effect of surface charge from these data indicated that maxi K channels sensed the surface potentials at a distance of $8{\sim}9\;{\AA}$ from the membrane surface. In addition, similar insulation distance ($7{\sim}9\;{\AA}$) of channel mouth from the bilayer surface charge was predicted by a 3-barrier-2-site model of energy profile for the permeation of $K^+$ ions. In conclusion, despite the differences in structure and fluidity of phospholipids in bilayers, the activities of maxi K channels in two charged membranes composed of PS or PI were strikingly similar and larger than those in bilayers of PE. These results suggest that the enhancement of conductance and $P_o$ of maxi channels is mostly due to negative charges in the phospholipid head groups.

• 한국응용과학기술학회지
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• 제39권5호
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• pp.720-726
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• 2022

형광 강도 변화를 이용하여 세포막의 위상에 따른 하이드록시부티르산 혼입이 생체막에 미치는 영향을 조사하였다. 구형 인지질 이중층인 소포(vesicle)가 이중 에멀젼 기술을 통해 각 층 단위로 제조되었다. 소포 내부의 수성에는 아미노나프탈렌트리술폰산디소듐(ANTS)이 캡슐화되었으며, 소광제(quencher)로 자일렌비스피리디늄브로마이드(DPX)를 소포가 분산된 버퍼에 포함시켰다. 형광 등급은 100% 형광으로서 DPX가 포함된 완충액의 소포와 0% 형광으로서 ANTS와 DPX의 혼합물이 포함된 완충액의 소포를 고려하여 조정되었다. 소포 용액에 하이드록시부티르산 혼입은 막 구조의 변화를 유도하였으며, 이러한 변화는 하이드록시부티르산 대 지질의 비율에서 소포의 각 층의 상과 관련이 있는 것으로 관찰된다. 관찰된 결과는 머리 그룹과 꼬리 그룹 모두의 패킹 붕괴에 대한 삼투압 및 체적 효과로 인해 소포의 안정성에 의존하는 것으로 보인다.

• Archives of Pharmacal Research
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• 제27권11호
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• pp.1177-1182
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• 2004

The effect of poly[2-methacryloyloxyethyl phosphorylcholine] (pMPC) on the skin permeation property was investigated by performing in vitro skin permeation study of a model drug, nicotinic acid (NA). Effect of pMPC polymer in donor solution on skin permeation rates was evaluated using side-by-side diffusion cells. Also, the structural alterations in the stratum corneum (SC), inter-lamellar bilayer (ILB) and dermis layers in pMPC-treated and -untreated skin sections were investigated with transmission electron microscopy (TEM). The permeation profile of NA without pMPC in donor solution showed biphasic mode: initial $1^{st} phase and 2^{nd}$ hydration phase. The sudden, more than 10-fold increase in flux from the initial steady state (43.5 $\mu g/cm^2$/hr) to the $2^{nd}$ hydration phase (457.3 $\mu g/cm^2$/hr) suggests the disruption of skin barrier function due to extensive hydration. The permeation profile of NA with 3% pMPC in the donor solution showed monophasic pattern: the steady state flux (10.9 $\mu g/cm^2$/hr) without abrupt increase of the flux. The degree of NA permeation rate decreased in a concentration-dependent manner of pMPC. TEM of skin equilibrated with water or 2% pMPC for 12 h showed that corneocytes are still cohesive and epidermis is tightly bound to dermis in 2% pMPC-treated skin, while wider separation between corneocytes and focal dilations in inter-cellular spaces were observed in water-treated skin. This result suggests that pMPC could protect the barrier property of the stratum corneum by preventing the disruption of ILB structure caused by extensive skin hydration during skin permeation study.

• Bulletin of the Korean Chemical Society
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• 제34권11호
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• pp.3223-3226
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• 2013

Spherical phospholipid-bilayers, vesicles, were prepared using the layer-by-layer double emulsion technique, which allows the bilayer to be formed asymmetrically. On the outer layer of the vesicles, the phospholipase D (PLD) reacted to convert phosphatidylcholine (PC) to phosphatidic acid (PA). The reaction induced the curvature change of the vesicles, which eventually led to rupture. The response time from the time of PLD injection to the time of rupture was measured against different vesicle curvatures and the outer layer phase, using the fluorescence intensity change of a pH-sensitive dye encapsulated within the vesicles. The effect of the vesicle curvature on the response was observed to be more significantly dramatic at the solid phase, compared to the liquid phase. Furthermore, in the solid phase, the response time was faster for 80 and 155 nm vesicles and, slower for 605 nm vesicles than similarly sized vesicles in the liquid phase vesicles. This difference in the response time was thought to result from the configuration determined by the phase difference and the PLD behavior.

• Archives of Pharmacal Research
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• 제15권2호
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• pp.152-161
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• 1992

Selective quenching of 1, 6-diphenyl-1, 3, 5-hexatriene (DPH) by trinitrophenyl groups was utilized to examine the transbilayer fluidity asymmetry of model membranes of phospholipids (SPMVPL) extracted from synaptosomal plasma membrane vesicles (SPMV). The polarization (P), anisotropy (r), limiting anisotropy $(r_\infty$), and order parameter (S) of DPH in the inner monolayer were 0.019, 0.014, 0.018, and 0.047, respectively, greater than calculated for the outer monolayer of SPMVPL. Selective quenching of DPH by trinitrophenyl groups was also utilized to examine the effects of n-alkanols on the individual monolayer structure of SPMVPL. n-Alkanols fluidized the hydrocarbon region of bulk SPMVPL and the potencies of n-alkanols up to 1-nonanon increased with carbon chain length. It appears that the potencies in bilayer fluidization increase by 1 order of magnitude as the carbon chain length increases by two carbon atoms. The cut-off phenomenon was reached at 1-decanol, where further increase in hydrocarbon length resulted in a decrease in pharmacological activity. The n-alkanols had greater fluidizing effects on the outer monolayer as compared to the inner monolayer of SPMVPL, even though these selective effects tended to become weaker as the carbon chain length increased. Thus, it has been proven that n-alkanols exhibit selective rather than nonselective fludizing effects within transbilayer domains of SPMVPL.

• The Korean Journal of Physiology and Pharmacology
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• 제7권3호
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• pp.125-130
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• 2003

The aim of this study was to provide a basis for studying the molecular mechanism of pharmacological action of chlorhexidine digluconate. Fluorescence polarization of n-(9-anthroyloxy)stearic acid was used to examine the effect of chlorhexidine digluconate on differential rotational mobility of different positions of the number of membrane bilayer phospholipid carbon atoms. The six membrane components differed with respect to 2, 3, 6, 9, 12, and 16-(9-anthroyloxy)stearic acid (2-AS, 3-AS, 6-AS, 9-AS, 12-AS and 16-AP) probes, indicating different membrane fluidity. Chlorhexidine digluconate increased the rate of rotational mobility of hydrocarbon interior of the cultured Porphyromonas gingivalis outer membranes (OPG) in a dose-dependent manner, but decreased the mobility of surface region (membrane interface) of the OPG. Disordering or ordering effects of chlorhexidine digluconate on membrane lipids may be responsible for some, but not all of its bacteriostatic and bactericidal actions.

• 한국식품영양과학회지
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• 제33권5호
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• pp.864-868
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• 2004

철분은 생체 내에서 이루어지는 거의 모든 대사에 필수적인 성분이지만, 식음에 포함된 철분의 양은 극히 적어서 철분강화에 대한 연구가 꾸준히 진행되어 왔다. 이에 따라， 철분공여물질을 함유한 리포즘을 이용하여 철분 강화 식품첨가제를 개발하였다. 철분공여물질로 ferrous sulfate와 hemin을 사용하였으며, 이러한 철분 함유 리포좀을 제조하는데 가장 큰 문제점은 ferrous sulfate의 자체 산화와 ferrous sul fate와 hemin으로 인한 리포좀의 지질산화로 지적되었다 또한, ferrous sulfate에 의한 리포좀의 산화 정도는 hemin의 경우보다 낮은 것으로 관찰되었다. Ferrous sulfate의 자동 산화를 억제하기 위하여 수용성 항산화제인 ascorbic acid가 첨가되었으나, 첨가된 ascorbic acid는 ferrous sulfate와 hemin을 함유한 리포좀의 산화를 억제시키지 못했으며, 오히려 ferrous sulfate에 의한 리포좀의 산화를 촉진시키는 것으로 관찰되었다 여기에 지용성 항산화제인 $\alpha$-tocopherol을 추가적으로 첨가함으로써, ferrous sulfate의 자동산화를 억제하고 hemin과 ferrous sulfate에 의한 리포좀의 산화가 억제된 철분 함유 리포좀이 제조되었다.

• 공업화학
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• 제9권3호
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• pp.457-461
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• 1998

본 연구에서는 베지클의 응용성을 확장하기 위한 기초연구로서, DPPC(dipalmitoylphosphaticylcholine)/Chol(Cholesterol)계에 불소화계면활성제($C_8F_{17}(CH_2)_2OCO-CH_2CH(SO_3Na)COO(CH_2)_2C_8F_{17}$, FS)/불소화지방산염 (($C_7F_{15}COONH_4$, FFS)을 여러 가지 비율로 변화시켜 첨가함으로써 베지클을 변형 제조하고, 그 물리화학적 성질을 비교측정하였다. 제조된 베지클계에 대한 제타-전위 측정(Zetamaster Particle Electrophoresis Analyzer) 결과, 균일한 전하의 베지클이 형성되는 것으로 확인되었고, 이는 FFS 분자가 FS와 DPPC 사이에서 보조계면활성제로 기능하여 DPPC와 FS의 각 성분이 동일 베지클내에 공존하도록 작용했기 때문으로 풀이된다. 베지클의 입자크기 결정에서는 FFS의 농도가 증가할수록 그 크기가 감소하고, 제타-전위도 음의 값으로 더욱 작아지는 것으로 나타났다. 베지클의 분산도 변화에서도 DPPC/Chol/FS/FFS계의 경우가 DPPC/Chol/FS계에 비해 안정성이 훨씬 큰 것으로 조사되었다. 베지클의 서방유출 실험에서도 막내에 포집된 MB(Methylene Blue)의 유출속도가 DPPC/Chol/FS/FFS계에서 감소된다는 결과를 얻었고, 이는 FFS의 첨가로 인해 막의 유동성이 감소되기 때문으로 풀이된다. 이 결과는 형광방출 측정의 $I_{Excimer}/I_{Monomer}$값에서도 확인되는 것으로 막의 미세점도가 FFS 첨가 베지클에서 증가되는 것으로 나타났다. 단백질로서 알부민에 대한 친화도를 비교한 결과에서는 DPPC/Chol/FS/FFS계의 친화도가 다소 낮아지는 것으로 나타났다. 이들 실험결과에 바탕하여 FFS의 첨가에 의해 DPPC/Chol/FS 베지클계는 더욱 안정하고 균일한 막의 구조를 갖는 리포솜을 형성한다는 결론을 얻었다.