磷脂微滴囊泡化的介观模拟

发展了一种非显示溶剂的粗粒化三粒子磷脂模型,该模型明确反映磷脂分子的双尾结构．模型分别采用变形的MIE作用势和Harmonic作用势描述分子间非成键和分子内成键相互作用,粗粒化力场参数通过拟合DPPC双分子层的结构和力学性质获得．该粗粒化模型成功重现了磷脂分子从随机初始态到双分子层和从盘状结构到囊泡的形成过程．应用该模型系统研究了球形和柱形磷脂微滴囊泡化的过程,结果表明此模型能有效地模拟介观尺度下复杂磷脂囊泡的形成及演化．     

多组分磷脂巨囊泡的相结构和胆固醇对微畴成长的影响

摘要：巨囊泡作为细胞的简化模型,其分相与出芽机理及动力学规律已引起许多领域科学家的关注。在富含胆固醇的典型生物膜体系如二棕榈酰磷脂酰胆碱DPPC(2-dihexadecanoyl-rac-glycero-3phosphocholine)/二油酰磷脂酰胆碱DOPC(dioleoyl-phosphatidylcholine)/胆固醇(Chol)的三组分形成的巨囊泡作为模型,从高温退火至低温会发生相分离,形成微畴。实验中借助荧光显微镜观察生物膜体系侧向分离的相结构图。实验发现,体系各组分的不同会影响磷脂膜的相结构和膜内微畴的成长,固定 DOPC/DPPC为1:1的前提下,微畴尺寸随着胆固醇参入量的增加而变大。最后运用理论进一步分析了微畴的成长机理。     

物理因素对磷脂酰胆碱单分子层影响的研究

利用LB技术,在不同的物理条件下对磷脂酰胆碱单分子成膜质量和分子的构象变化进行了研究,在此础上对磷脂酰胆碱和胆固醇组成的复合膜的相变过程进行了研究,利用AFM对磷脂酰胆碱分子LB膜的分子构象和二维排布进行了表征.研究结果表明,物理因素温度、pH、浓度及胆固醇对于气液界面上磷脂酰胆碱的分子构象的结构有较大的影响.通过选择合适的条件能够得到具有特殊形态和结构的磷脂酰胆碱LB膜.     

原子力显微镜研究DPPC磷脂多层膜结构与力学性能

应用原子力显微镜(AFM)首先研究了在蔗糖溶液中二棕榈酰磷脂酰胆碱(DPPC)磷脂双层膜的结构,分析了其力学性能;其次,研究了在纯水中、CaCl2溶液中的DPPC磷脂多层膜的结构特性和杨氏模量.实验结果表明,在CaCl2溶液中DPPC多层膜的水层厚度大于在纯水中厚度,在CaCl2溶液中多层膜的杨氏模量变小.     

原子力显微镜观察脂胞囊形貌结构及稳定性

用原子力显微镜(AFM)观察了液晶态脂胞囊形貌结构和稳定性.实验结果显示,对于1,2-二油酸甘油-3-磷脂酰乙醇胺(DOPE)超分子聚集体,在扫描区域内,可观察到一些大小不同球形颗粒;对于1,2-二油酸甘油-3-磷脂酸乙醇胺(DOPE)和1,2-二油酸甘油-3-磷脂酰胆碱(DOPC)混合的超分子聚集体,则不仅观察到类似脂胞囊的球形颗粒,而且也观察到了双层膜结构,其厚度约为5～6 nm.     

用小角X射线散射法研究CH_2Cl_2,CHCl_3和CCl_4对磷脂酰胆碱液晶态结构的影响机理

本文采用小角X射线散射法分别研究CH_2Cl_2,CHCl_3和CCl_4对磷脂酰胆碱液晶态结构的影响机理,通过比较得知,CH_2Cl_2,CHCl_3和CCl_4对磷脂酰胆碱液晶态结构影响的不同之处,主要是它们空间旋转电子云密度分布不同所致,空间旋转电子云密度分布呈球状或椭球状的物质都有使磷脂酰胆碱液晶形成片层六角形的机理,呈圆锥状的物质有诱发磷脂酰胆碱液晶形成六角形H_(Ⅱ)相的机理。     

多聚赖氨酸诱导的负电性磷脂巨囊泡形变

利用荧光显微技术表征了多聚赖氨酸诱导的负电性磷脂巨囊泡的动力学响应行为.研究发现,多聚赖氨酸可吸附至二油酰磷脂酰胆碱和二油酰磷脂酸混合磷脂巨囊泡的表面,诱导其发生粘连、出"绳"及破裂现象.分析认为,在低盐环境中,膜形变由多聚赖氨酸吸附于二油酰磷脂酸富集区引起的膜两叶应力不对称,以及静电相互作用等因素产生.研究结果对基于聚合物-巨囊泡体系的药物输运控释、细胞形变、微控反应和基因治疗等方面的研究提供有价值的支持.     

用小角X射线散射法研究CH2Cl2,CHCl3和CCl4对磷脂酰胆碱液晶态结构的影响机理

本文采用小角X射线散射法分别研究CH₂Cl₂,CHCl₃和CCl₄对磷脂酰胆碱液晶态结构的影响机理,通过比较得知,CH₂Cl₂,CHCl₃和CCl₄对磷脂酰胆碱液晶态结构影响的不同之处,主要是它们空间旋转电子云密度分布不同所致,空间旋转电子云密度分布呈球状或椭球状的物质都有使磷脂酰胆碱液晶形成片层六角形的机理,呈圆锥状的物质有诱发磷 关键词：     

超支化聚合物组装体质子捕获能力的初步研究

用荧光光谱法研究超支化聚合物组装体的质子捕获能力,共三种组装体结构,HBPO-star-PDMAEMA胶束、BP囊泡(HBPO-star-PEO)和BPD囊泡(HBPO-star-PEO和HBPO-star-PDMAEMA共组装)。结果发现由于质子海绵型结构PDMAEMA的存在,HBPO-star-PDMAEMA胶束和BPD囊泡在叔胺质子化过程中,具有较强的质子捕获能力,而Pyranine包埋于BP囊泡内与其在水中的光谱随环境pH值变化无明显差异。因而,通过调节HBPO-star-PEO和HBPO-star-PDMAEMA的比例,我们可以制备得到一系列质子捕获能力的超支化聚合物囊泡。     

磷脂酰胆碱SUV的NMR研究

磷脂酰胆碱(PC)是人和动物血液中大量存在的一类具有重要生物功能的磷脂,它和鞘磷脂一起形成了不同大小的密度脂蛋白,对血液中胆固醇等分子的转运和代谢起着至关重要的作用.脂蛋白中磷脂的组成和形态变化与某些疾病,如动脉粥样硬化、癌症和老年痴呆等的发生和发展密切相关,因此研究磷脂的组成形态将有助于明确磷脂的生物化学作用.该文采用一维(1D)和二维(2D)NMR技术对PC所形成的SUV(small unilamellar vesicle)结构进行了分析,通过对PC磷脂头部氮甲基的检测分析,发现PC所形成的SUV为较为稳定的双层结构,这表明通常的磷脂脂蛋白可能是一种双层膜结构,而非通常所认为的单层结构.     

Interaction of dipalmitoyl phosphatidylcholine (DPPC) liposomes and insulin

Insulin, a peptide that has been used for decades in the treatment of diabetes, has well-defined properties and delivery requirements. Liposomes, which are lipid bilayer vesicles, have gained increasing attention as drug carriers which reduce the toxicity and increase the pharmacological activity of various drugs. The molecular interaction between (uncharged lipid) dipalmitoyl phosphatidylcholine (DPPC) liposomes and insulin has been characterized by using Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction. The characteristic protein absorption band peaks, Amide I (at about 1660?cm^?1) and Amide II band (at about 1546?cm^?1) are potentially reduced in the liposome insulin complex. Wide-angle x-ray scattering measurements showed that the association of insulin with DPPC lipid of liposomes still maintains the characteristic DPPC diffraction peaks with almost no change in relative intensities or change in peak positions. The absence of any shift in protein peak positions after insulin being associated with DPPC liposomes indicates that insulin is successfully forming complex with DPPC liposomes with possibly no pronounced alterations in the structure of insulin molecule.     

FTIR法研究氨基酸稀土配合物与磷脂的相互作用

利用傅里叶变换红外光谱（ＦＴＩＲ）法研究了不同类型氨基酸及其配合物与二棕榈酰磷脂酰胆碱脂质体浇铸膜的相互作用。结果表明，在有氨基酸存在下，二棕榈酰磷脂酰胆碱的红外谱图没有明显的变化；氨基酸稀土配合物与磷脂极性头基团有静电相互作用，作用的程度随稀土离子、氨基酸及稀土同氨基酸配比的不同而有所不同。     

Permeation of nanocrystals across lipid membranes

Biological membranes are one of the major structural elements of cells, and play a key role as a selective barrier and substrate for many proteins that facilitate transport and signaling processes. Understanding the structural and mechanical properties of lipid membranes during permeation of nanomaterials is of prime importance in determining the toxicity of nanomaterials to living cells. It has been shown that the interaction between lipid membranes and nanomaterials and the disruption of lipid membranes are often determined by physicochemical properties of nanomaterials, such as size, shape and surface composition. In this work, molecular dynamic simulations were carried out using various sizes of nanocrystals as a probe to explore the transport of nanomaterials across dipalmitoylphosphatidylcholine (DPPC) bilayers and the changes in the structural and mechanical properties of DPPC bilayers during the permeation. A coarse-grained model was used to provide insight at large time and length scales. In this work, an external driving force helps the nanocrystals across the lipid bilayer. The minimum forces needed to penetrate the model membrane and the interaction of nanocrystals and lipid bilayers were investigated in simulations. The elastic and dynamic properties of lipid bilayers, including the local and bulk properties during the permeation of the nanocrystals, which are of considerable fundamental interest, were also studied. The findings described will lead to better understanding of nanomaterial–lipid membrane interactions and the mechanical and dynamic properties of lipid membranes under permeation.     

Microscopical investigations of nisin-loaded nanoliposomes prepared by Mozafari method and their bacterial targeting

Nanoencapsulation may improve activity of protein or polypeptide antimicrobials against a variety of microorganisms. In this study, nanoliposomes prepared from different lipids (Phospholipon 90H, Phospholipon 100H, dipalmitoylphosphatidylcholine (DPPC), stearylamine (SA), dicetyl phosphate (DCP) and cholesterol) by a new, non-toxic and scalable method, were tested for their capacity to encapsulate nisin Z and target bacteria (Bacillus subtilis and Pseudomonas aeruginosa). Factors affecting the entrapment efficiency (charge and cholesterol concentration in the vesicles) and stability of nanoliposomes were assessed. The nanoliposomes and their bacterial targeting were visualised, using different microscopes under air and liquid environments. Nisin was entrapped in different nanoliposomes with encapsulation efficiencies (EE) ranging from 12% to 54%. Anionic vesicles possessed the highest EE for nisin while increase in cholesterol content in lipid membranes up to 20% molar ratio resulted in a reduction in EE. Stability of nanoliposome-encapsulated nisin was demonstrated for at least 14 months at 4 °C (DPPC:DCP:CHOL vesicles) and for 12 months at 25 °C (DPPC:SA:CHOL vesicles).     

Preparation of Long-Chain Fatty Acyl-Grafted Chitosan in an Ionic Liquid and Their Self-Assembled Micelles in Water

Controllable synthesis of self-associating lauroyl-grafted chitosan with high molecular weight was realized in ionic liquid reacting media. The obtained lauroyl chitosan with different grafting degrees was characterized by FT-IR and ¹H-NMR spectra. The chitosan derivatives were able to self-assemble into spherical polymeric micelles in water. The onset of micellization or critical micelle concentration (CMC) was estimated by fluorescence spectroscopy. The hydrodynamic diameters of various lauroyl chitosan micelles, determined by dynamic light scattering, were in the range of 122–295 nm. The morphology of the micelles was observed by scanning electron microscopy. The self-assembly behavior and the size of the assembled micelles of lauroyl chitosan were affected by the grafting degree of lauroyl groups. Generally, higher grafting degree resulted in lower CMC and smaller micelle size. These lauroyl chitosan nano-micelles may have potential applications in biological and medical fields.     

Influence of salicylic acid on the biophysical properties of dipalmitoyl phosphatidylcholine vesicles

The effect of the keratolytic drug salicylic acid (SA) on the thermotropic behaviour, and dynamics of dipalmitoyl phosphatidyl choline (DPPC)–water/buffer pH?7.4 vesicles was studied using DSC and ¹H NMR. In both systems, incorporation of SA in DPPC bilayer causes a significant depression in the transition temperature of both the pre-transition (PT) and the gel-to-liquid crystalline (CM) transition. The presence of the drug reduces the cooperativity of both the PT and CM transitions. These findings indicate that SA is bound strongly to the lipid bilayer leading to increased membrane fluidity. The DPPC vesicles incorporated with high drug concentration show phase segregation. One of the interesting findings in this study is the formation of a more ordered high temperature gel (L_β2) phase when the SA-doped DPPC dispersion is prepared at physiological pH. The effect of inclusion of cholesterol in the SA-free and SA-doped DPPC dispersion was also studied.     

A comparative electron spin resonance investigation on the effects of barbital and tetracaine on DPPC multilayers

We have performed a comparative spin label ESR investigation of the effects of tetracaine and barbital on DPPC multilamellar vesicles (MLV). The investigation has been carried out both at pH 5.0 and 9.0, by using the spin labels 5-SASL, 5-PCSL and DTBN as magnetic probes. Information on the thermotropic properties of the multilamellae, such as molecular orientational degree of order, packing density and permeability has been obtained. The results show that tetracaine induces fluidization of the phospholipid bilayers, reduction of the molecular packing density and downward shift of the main phase transition temperature of the multilayers of DPPC. Moreover, increasing concentrations of tetracaine are able to induce the lyotropic gel → liquid-crystal phase transition at a fixed temperature, in particular at pH 5.0. Barbital, instead, is much less effective than tetracaine in perturbing the membrane properties. In fact, the most important effect observed is only a progressive fluidization of the DPPC bilayers especially at pH 9.0. The dependence on pH of the effects observed suggests that the anesthetic molecules locate at different position in the lipid bilayers depending on the protonation state of both molecules.     

Determination of the rate of rapid lipid transfer induced by poly(ethylene glycol) using the SLM Fourier transform phase and modulation spectrofluorometer

Rate constants were determined for the transfer of the fluorescent lipid probe 1-palmitoyl-2-[[2-[4-(6-phenyl-trans-1,3,5-hexatrienyl)phenyl]ethyl] oxy]carbonyl]-3-sn-phosphatidylcholine (DPHpPC) between large, unilamellar extrusion vesicles composed either of dipalmitoyl phosphatidylcholine (DPPC) or of DPPC mixed with a small amount (0.5 mol%) of lyso phosphatidylcholine (Lyso PC). Transfer of the lipid probe in the presence of varying concentrations of poly(ethylene glycol) (PEG) was monitored using the SLM 48000-MHF Multi-Harmonic Fourier Transform phase and modulation spectrofluorometer to collect multifrequency phase and modulation fluorescence data sets on a subsecond time scale. The unique ability of this instrument to yield accurate fluorescence lifetime data on this time scale allowed transfer to be detected in terms of a time-dependent change in the fluorescent lifetime distribution associated with the lipid-like DPHpPC probe. This probe demonstrates two short fluoresence decay times (ca. 1.1–1.4 and 4.3–4.8 ns) in a probe-rich environment but a single long lifetime (ca. 7 ns) in a probe-poor environment. A simple two-state model for initial lipid transfer was used to analyze the multifrequency data sets collected over a 4-s time frame to obtain the time rate of change of the concentrations of donor and acceptor probe populations following rapid mixing of vesicles with PEG. The ability to measure fluorescence lifetimes on this time scale has allowed us to show that the of rate of lipid transfer increased dramatically at 35% PEG in both fusing and nonfusing vesicle systems. These results are interpreted in terms of a distinct interbilayer structure associated with intimate bilayer contact induced by high and potentially fusogenic concentrations of PEG.     

Miscibility phase diagrams of giant vesicles containing sphingomyelin

Saturated sphingomyelin (SM) lipids are implicated in lipid rafts in cell plasma membranes. Here we use fluorescence microscopy to observe coexisting liquid domains in vesicles containing SM, an unsaturated phosphatidylcholine lipid (either DOPC or POPC), and cholesterol. We note similar phase behavior in a model membrane mixture without SM (DOPC/DPPC/Chol), but find no micron-scale liquid domains in membranes of POPC/PSM/Chol. We delineate the onset of solid phases below the miscibility transition temperature, and detail indirect evidence for a three-phase coexistence of one solid and two liquid phases.