Hall S M, Haworth S G
Developmental Vascular Biology and Pharmacology Unit, Institute of Child Health, London, United Kingdom.
Am J Physiol. 1996 Mar;270(3 Pt 1):L435-45. doi: 10.1152/ajplung.1996.270.3.L435.
The efficacy of preservation fluids on the cytoskeleton and contractile function of porcine pulmonary arterial smooth muscle (SM) cells during cooling and rewarming was evaluated, using EuroCollins solution (EC), University of Wisconsin solution (UW), Marshall's solution (MS), and tissue culture growth medium (GM). Functional studies included passive distensibility and contraction to prostaglandin F2a (PGF2a) in arterial rings and wrinkling of silicone membranes by cooled-rewarmed cultured SM cells. Immunofluorescence measurements were made of actin brightness in cooled arterial rings. Cultured SM monolayers were stained with antibodies to SM alpha-actin, SM myosin, and tubulin. In cooling, all solutions resulted in increased arterial distensibility, whereas EC and MS reduced cell wrinkling. With the use of all solutions, actin cables thinned, myosin filaments dissociated, and microtubules depolymerized. During rewarming, resistance to imposed tension increased in all arterial rings. After GM,ED, and MS preservation, contraction to PGF2a increased. Wrinkling increased and actin-myosin cables shortened after GM and EC; after UW, wrinkling decreased and actin-myosin cables thinned. No recovery occurred after MS. Thus the type of preservation solution influenced contractility during preservation and after rewarming. The absence of spontaneous contraction in cells cooled in UW may be advantageous.
使用EuroCollins溶液(EC)、威斯康星大学溶液(UW)、马歇尔溶液(MS)和组织培养生长培养基(GM),评估了保存液对猪肺动脉平滑肌(SM)细胞在冷却和复温过程中细胞骨架和收缩功能的影响。功能研究包括动脉环的被动扩张性和对前列腺素F2a(PGF2a)的收缩反应,以及冷却复温后的培养SM细胞对硅膜的皱缩作用。通过免疫荧光测量冷却动脉环中肌动蛋白的亮度。用抗平滑肌α-肌动蛋白、平滑肌肌球蛋白和微管蛋白的抗体对培养的SM单层细胞进行染色。在冷却过程中,所有溶液均导致动脉扩张性增加,而EC和MS减少了细胞皱缩。使用所有溶液时,肌动蛋白丝变薄,肌球蛋白丝解离,微管解聚。在复温过程中,所有动脉环对施加张力的阻力增加。在GM、ED和MS保存后,对PGF2a的收缩反应增强。GM和EC后皱缩增加,肌动蛋白-肌球蛋白丝缩短;UW后,皱缩减少,肌动蛋白-肌球蛋白丝变薄。MS后未出现恢复。因此,保存液的类型影响保存期间和复温后的收缩性。在UW中冷却的细胞中无自发收缩可能是有利的。