本研究利用螢光顯微鏡分析法（fluorescence microscopy, FM）配合表面壓-面積等溫線的量測，探討於連續來回壓縮-擴張界面條件下，牛血漿白蛋白（bovine serum albumin, BSA）對氣/液界面上dipalmitoyl phosphatidylcholine（DPPC）/1-palmitoyl-2-oleoyl phosphatidylglycerol（POPG）（莫耳比3:1）混合單分子層行為的影響。在BSA吸附分子層存在之氣/液界面上分佈DPPC分子後，表面壓-相對面積遲滯曲線與DPPC/BSA混合分子層的表面形態顯示，當混合分子層被壓縮至緊密狀態時，BSA分子會被DPPC分子擠出界面，而在BSA分子被擠出界面的同時，會造成界面上自由DPPC分子的損失。在後續的界面擴張階段中，表面壓的回升推測是BSA分子的再吸附所造成的。此外，在DPPC/BSA混合分子層的壓縮-擴張過程中，FM影像顯示BSA可能與DPPC作用形成複合物。由以上結果推斷，於連續來回壓縮-擴張界面條件下，BSA的存在會減少氣/液界面上自由DPPC分子的數量，使得DPPC的動態界面活性受到抑制。
在BSA吸附分子層存在之氣/液界面上分佈POPG分子後，於連續來回壓縮-擴張界面條件下，部分POPG分子會隨BSA分子離開氣/液界面，導致界面上自由POPG分子的損失，使得POPG的動態界面活性受到抑制。自由POPG分子的損失，可能與帶負電的BSA分子和帶負電頭基的POPG分子之靜電排斥作用，或POPG分子之碳氫鏈與BSA分子之疏水結構的疏水作用有關。因此於連續來回壓縮-擴張界面條件下，當BSA分子離開界面時也造成POPG分子的損失。
在BSA吸附分子層存在之氣/液界面上同時分佈DPPC及POPG分子後，發現DPPC/POPG/BSA混合分子層與DPPC/BSA混合分子層的動態界面行為非常相似，顯示POPG在界面上的分率明顯減少。這可能是因為BSA與POPG的作用較強，因此於連續來回壓縮-擴張界面條件下，BSA分子會選擇性地將POPG分子移出界面，使界面上的自由POPG分子嚴重損失，因而出現由DPPC與BSA主導的動態界面行為。

The effects of bovine serum albumin (BSA) on the mixed dipalmitoyl phosphatidylcholine (DPPC)/1-palmitoyl-2-oleoyl phosphatidylglycerol (POPG) (molar ratio 3:1) monolayer behavior at cyclic air/liquid interfaces were investigated by the fluorescence microscopy (FM) with the measurements of surface pressure-area isotherms. With spreading DPPC molecules onto the air/liquid interface with an adsorbed BSA layer, it was revealed from the surface pressure-relative area hysteresis curves and the morphology of mixed DPPC/BSA layers that BSA molecules were expelled from the interface by DPPC molecules as the mixed layer was compressed to a condensed state. The squeeze-out of BSA molecules from the interface would induce the loss of free DPPC molecules at the interface. The increased surface pressure during the following interface expansion stage was probably due to the readsorption of BSA molecules. Furthermore, during the compression-expansion process of mixed DPPC/BSA layers, FM images indicated that BSA might interact with DPPC to form complexes. The results suggested that under the condition of continuous interface compression-expansion, the presence of BSA would decrease the number of free DPPC molecules at the air/liquid interface, resulting in the inhibited dynamic surface activity of DPPC.
After spreading POPG molecules onto the air/liquid interface with an adsorbed BSA layer, under the condition of continuous interface compression-expansion, it was found that part of POPG molecules would leave the air/liquid interface with BSA molecules, resulting in the loss of free POPG molecules at the interface and the inhibited dynamic surface activity of POPG. The loss of free POPG molecules was probably related to the electrostatic repulsion between negatively charged BSA molecules and headgroups of POPG molecules, or to the hydrophobic interactions between the hydrocarbon chains of POPG molecules and the hydrophobic part of BSA molecules. Therefore, under the condition of continuous interface compression-expansion, when BSA molecules left the interface, it also caused the loss of POPG molecules.
After simultaneously spreading DPPC and POPG molecules onto the interface with an adsorbed BSA layer, it was found that the dynamic interface behavior of the mixed DPPC/POPG/BSA layer was very similar to that of the mixed DPPC/BSA layer, indicating that the POPG fraction at the interface was significantly decreased. This is probably caused by the stranger interaction between BSA and POPG. As a result, under the condition of continuously interface compression-expansion, POPG molecules were selectively removed from the interface by BSA and the amount of free POPG molecules at the interface was dramatically decreased, resulting in the dynamic interface behavior dominated by DPPC and BSA.

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