基因载体材料聚乙烯亚胺与磷脂酰胆碱脂质体的相互作用对膜结构的影响

doi:10.7503/cjcu20170273

摘要/Abstract

摘要：

采用动态光散射、荧光光谱、 zeta电位测定和等温滴定量热技术分析了分子量分别为25000, 10000和1800的聚乙烯亚胺(PEI)与二油酰磷脂酰胆碱(DOPC)脂质体的相互作用及其对脂质体膜内环境极性和膜通透性的影响. 结果表明, PEI通过氨基与DOPC的磷脂基团和胆碱基团产生氢键或范德华作用, 从而与脂质体结合形成复合物; 低浓度PEI(<0.075 mg/mL)导致DOPC脂质体的聚集和表面电位的增加, 但未引起脂质体膜融合和表面电位反转; 进一步增加PEI的浓度对脂质体表面电位的影响很小, 而结合在表面的PEI分子链之间的排斥作用阻碍了脂质体聚集. PEI分子与DOPC脂质体的结合降低了脂质分子碳氢链的堆积密度和脂质体膜内环境的疏水性, 从而增强了钙黄绿素和槲皮素在脂质体膜中的通透性. PEI与DOPC脂质体的相互作用具有明显的分子尺寸效应, 增大PEI的分子量可以增强与脂质体的相互作用及对脂质体膜结构的影响.

关键词: 聚乙烯亚胺, 磷脂酰胆碱, 脂质体, 相互作用, 膜结构

Abstract:

To get a better understanding on the molecular basis of the cytotoxicity of PEI which has been considered as “golden standard” for polymeric gene delivery carriers. Dynamic light scattering, fluorescence spectra, zeta-potential measurement and isothermal titration calorimetry were conducted to reveal the mechanism of interaction between PEIs(average molecular weight of 25000, 10000 and 1800) and 1,2-dioleoyl-sn-glycero-3-phosphatidylcholine(DOPC) liposome. The influence on the polarity of microenvironment and the permeability of liposome bilayer were also investigated. The result showed that PEI bound to DOPC vesicles via hydrogen bond or van Waals interactions between the amide groups and the phosphorylcholine heads. The complex formation with PEI induced aggregation and increase in zeta potential of liposomes at low PEI concentration up to 0.075 mg/mL. A further increase in PEI concentration made little change on the surface potential, however reduced the aggregation of the vesicle due to the repulsion between the adsorbed PEI chains. PEI binding decreased the packing density of hydrocarbon chain of lipid molecules and the hydrophobicity in the bilayer membrane, and thus resulted in an enhanced permeability of calcein and quercetin through the membrane. The polymer size played an important role in PEI-DOPC liposome interaction. PEI with higher molecular weight was more favorable to interact with DOPC and more efficient to perturb the structural properties of the membrane.

Key words: Polyethyleneimine, Phosphatidylcholine, Liposome, Interaction, Membrane structure

中图分类号:

O648

TrendMD:

韦艳珊, 韦邦帜, 黄爱民, 马林. 基因载体材料聚乙烯亚胺与磷脂酰胆碱脂质体的相互作用对膜结构的影响. 高等学校化学学报, 2017, 38(12): 2271.

WEI Yanshan, WEI Bangzhi, HUANG Aimin, MA Lin. Membrane Structure Alteration of DOPC Liposome Induced by Interaction with Gene Carrier Polyethyleneimine^†. Chem. J. Chinese Universities, 2017, 38(12): 2271.

图/表 9

Fig.1 PEI concentration dependence of hydrodynamic diameter(Dhy) of DOPC LUVs in PBS cDOPC=50 μmol/L.

Fig.2 Fluorescence spectra of DOPC LUVs labeled with NBD-PE and Rh-PE in PBS(pH=7.4) containing 0(a), 0.05(b) and 0.1(c) mg/mL PEI25kInsert: relative intensity of fluorescence emission of NBD-PE and Rh-PE(F520/F590) in DOPC LUVs in PBS containing different concentrations of PEI25k. λex=463 nm, c(DOPC)=50 μmol/L.

Fig.3 Variation of zeta potential of DOPC LUVs in PBS(pH=7.4) containing PEIs c(DOPC)=50 μmol/L.

Fig.4 ITC profiles for titration of PEI25k into DOPC LUVs in PBS solution at 25 ℃ and pH=7.4(A) Exothermic heat release upon injection of PEI25k into DOPC LUVs dispersion; (B) integrated heat data associated with the interaction of PEI25k and DOPC LUVs. c(PEI25k)=5.0 mg/mL, c(DOPC)=750 μmol/L.

Fig.5 Schematic representation of concentration-dependent interaction of PEI with DOPC LUVs

Fig.6 PEI concentration dependence of ANS fluorescence emission maximum in PBS(pH=7.4)in the absence(dash line) and presence(solid line) of DOPC LUVsλex=388 nm; c(ANS)=2.5 μmol/L; c(DOPC)=50 μmol/L.

Fig.7 Time course leakage of calcein from DOPC LUVs in PBS containing PEI25k c(DOPC)=2.5 μmol/L.

Fig.8 PEI concentration dependence of maximum leakage of calcein from DOPC LUVs c(DOPC)=2.5 μmol/L.

Fig.9 PEI concentration dependence of fluorescence intensity of quercetin at 564 nm(F564)(A) In the absence(solid line) and presence(dash line) of DOPC LUVs in PBS(pH=7.4); (B) difference in the fluorescence emission in the absence and presence of DOPC LUVs. λex=465 nm, c(DOPC)=50 μmol/L, c(Que)=5 μmol/L.

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