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在空气/水界面含有两亲性肽的人工肺表面活性剂混合物的朗缪尔单分子层:新制剂与Surfacten(表面活性剂TA)的比较

Langmuir monolayer of artificial pulmonary surfactant mixtures with an amphiphilic peptide at the air/water interface: comparison of new preparations with surfacten (Surfactant TA).

作者信息

Nakahara Hiromichi, Lee Sannamu, Sugihara Gohsuke, Chang Chien-Hsiang, Shibata Osamu

机构信息

Division of Biointerfacial Science, Graduate School of Pharmaceutical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.

出版信息

Langmuir. 2008 Apr 1;24(7):3370-9. doi: 10.1021/la703180x. Epub 2008 Mar 4.

Abstract

Interfacial behavior was studied on the pulmonary lipid mixture containing a newly designed amphiphilic alpha-helical peptide (Hel 13-5) that consists of 13 hydrophobic and 5 hydrophilic amino acid residues. Moreover, the data obtained were compared with those of commercially available Surfacten (Surfactant TA) which has been clinically used for neonatal respiratory distress syndrome (NRDS) in Japan. Surface pressure (pi)-A and surface potential (DeltaV)-area (A) isotherms were measured for our synthetic preparations and Surfacten. Herein, a mixture of dipalmitoylphosphatidylcholine (DPPC)/egg-phosphatidylglycerol (PG)/palmitic acid (PA) (68:22:9 by weight) was used as the constituent of basic preparations. Monolayers were spread on 0.02 M Tris buffer (pH 7.4) with 0.13 M NaCl at the air/liquid interface, and the surface behavior was investigated by employing the Wilhelmy method, an ionizing electrode method, and fluorescence microscopy (FM). Cyclic compression and expansion isotherms of the prepared materials (or products) (DPPC/PG/PA/Hel 13-5) were examined to confirm the spreading and respreading ability. For the prepared products, a plateau region exists on pi-A and DeltaV-A isotherms at approximately 42 mN m(-1), indicating that Hel 13-5 is squeezed out of surface monolayers together with fluid components (PG) upon lateral compression. That is, the squeeze-out phenomenon induces a 2D-3D phase transformation. In particular, the inclination of the pi-A isotherms at X(Hel 13-5) = 0.1 in the plateau region was almost zero irrespective of the molecular area. As proposed in the earlier report (Nakahara, H.; Lee, S.; Sugihara, G.; Shibata, O. Langmuir 2006, 22, 5792-5803), an observed refluorescence phenomenon was discussed for FM measurements. This phenomenon provides evidence of the squeeze-out motion with fluid molecules. Furthermore, the cyclic pi-A and DeltaV-A isotherms show larger hysteresis areas and better respreading abilities in comparison with the previous ternary systems (DPPC/PG/Hel 13-5 and DPPC/PA/Hel 13-5) that are very important properties in pulmonary functions. FM photographs and the temperature dependence of pi-A and DeltaV-A isotherms suggest that the phase behavior of the present preparation product is very similar to that of Surfacten in terms of the domain size and in parameters such as collapse pressures, maximum DeltaV values, and so on. These results demonstrate that PG and PA even in the present preparations work well for compression-expansion cycling as is the case in the previous ternary systems, and the present preparations show comparable properties to Surfacten in vitro.

摘要

研究了包含一种新设计的两亲性α-螺旋肽(Hel 13-5)的肺脂质混合物的界面行为,该肽由13个疏水氨基酸残基和5个亲水氨基酸残基组成。此外,将获得的数据与在日本临床上用于新生儿呼吸窘迫综合征(NRDS)的市售Surfacten(表面活性剂TA)的数据进行了比较。对我们的合成制剂和Surfacten测量了表面压力(π)-面积(A)和表面电位(ΔV)-面积(A)等温线。在此,使用二棕榈酰磷脂酰胆碱(DPPC)/卵磷脂酰甘油(PG)/棕榈酸(PA)(重量比为68:22:9)的混合物作为基础制剂的成分。单分子层铺展在空气/液体界面处含有0.13 M NaCl的0.02 M Tris缓冲液(pH 7.4)上,并采用Wilhelmy法、电离电极法和荧光显微镜(FM)研究表面行为。检查了制备材料(或产品)(DPPC/PG/PA/Hel 13-5)的循环压缩和膨胀等温线,以确认铺展和再铺展能力。对于制备的产品,在π-A和ΔV-A等温线上大约42 mN m⁻¹处存在一个平台区域,这表明在横向压缩时Hel 13-5与流体成分(PG)一起被挤出表面单分子层。也就是说,挤出现象诱导了二维-三维相变。特别是,在平台区域中X(Hel 13-5)= 0.1时,π-A等温线的倾斜度几乎为零,与分子面积无关。如早期报告(Nakahara, H.; Lee, S.; Sugihara, G.; Shibata, O. Langmuir 2006, 22, 5792 - 5803)中所提出的,对FM测量中观察到的再荧光现象进行了讨论。这种现象为流体分子的挤出运动提供了证据。此外,与先前的三元体系(DPPC/PG/Hel 13-5和DPPC/PA/Hel 13-5)相比,循环π-A和ΔV-A等温线显示出更大的滞后面积和更好的再铺展能力,这些在肺功能中是非常重要的特性。FM照片以及π-A和ΔV-A等温线的温度依赖性表明,就畴尺寸以及诸如塌陷压力、最大ΔV值等参数而言,本制备产品的相行为与Surfacten非常相似。这些结果表明,即使在本制剂中,PG和PA在压缩-膨胀循环中也能很好地发挥作用,就像在先前的三元体系中一样,并且本制剂在体外表现出与Surfacten相当 的性质。

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