CAJ | 학술논문

目的：研究两种磷酸二酯酶（PDE）抑制剂对体外培养的大鼠脑微血管内皮细胞屏障功能的影响及机制。方法以原代方法培养大鼠脑微血管内皮细胞并传代，将第5代细胞随机分为对照组、西洛他唑组、双嘧达莫组、西洛他唑联合蛋白激酶（PKA）抑制剂 H89组、双嘧达莫联合 H89组，共5组，各组均模拟“缺血再灌注”过程，之后分别添加相应的药物，分析内皮细胞纤维肌动蛋白分布、紧密连接蛋白5的表达、对白蛋白和葡聚糖的通透性，以确定各种药物对大鼠脑微血管内皮细胞屏障特性的作用。结果西洛他唑和双嘧达莫改变了纤维肌动蛋白的分布。西洛他唑增加了紧密连接蛋白5的表达，免疫印迹定量显示为对照组的（218．2±39．3）％（P ＜0．01）。西洛他唑和双嘧达莫降低了内皮细胞对白蛋白的通透性，分别为（0．869±0．15）×10-6 cm/s（P ＜0．05）和（0．65±0．11）×10-6 cm/s（P ＜0．01）；蛋白激酶抑制剂 H89可基本逆转西洛他唑对白蛋白通透性降低的作用（P ＜0．05）；西洛他唑、西洛他唑联合H89降低了内皮细胞对葡聚糖的通透性（P ＜0．05），而双嘧达莫不能（P ＞0．05）。结论西洛他唑和双嘧达莫两种PDE 有增强大鼠脑微血管内皮细胞屏障特性，但具体作用及可能的机制不同，其中 PKA 介导下的西洛他唑作用更明显。
목적：연구량충린산이지매（PDE）억제제대체외배양적대서뇌미혈관내피세포병장공능적영향급궤제。방법이원대방법배양대서뇌미혈관내피세포병전대，장제5대세포수궤분위대조조、서락타서조、쌍밀체막조、서락타서연합단백격매（PKA）억제제 H89조、쌍밀체막연합 H89조，공5조，각조균모의“결혈재관주”과정，지후분별첨가상응적약물，분석내피세포섬유기동단백분포、긴밀련접단백5적표체、대백단백화포취당적통투성，이학정각충약물대대서뇌미혈관내피세포병장특성적작용。결과서락타서화쌍밀체막개변료섬유기동단백적분포。서락타서증가료긴밀련접단백5적표체，면역인적정량현시위대조조적（218．2±39．3）％（P ＜0．01）。서락타서화쌍밀체막강저료내피세포대백단백적통투성，분별위（0．869±0．15）×10-6 cm/s（P ＜0．05）화（0．65±0．11）×10-6 cm/s（P ＜0．01）；단백격매억제제 H89가기본역전서락타서대백단백통투성강저적작용（P ＜0．05）；서락타서、서락타서연합H89강저료내피세포대포취당적통투성（P ＜0．05），이쌍밀체막불능（P ＞0．05）。결론서락타서화쌍밀체막량충PDE 유증강대서뇌미혈관내피세포병장특성，단구체작용급가능적궤제불동，기중 PKA 개도하적서락타서작용경명현。
Objective To analyze the effects of two phosphodiesterase(PDE)inhibitors on brain microvascular endothelial cells (HBECs)barrier function in cultured rat.Methods In primary cultured rat brain microvascular endothelial cells and passage,the fifth generation of cells were randomly divided into positive control group,cilostazol group,dipyridamole group,cilostazol combined with protein kinase inhibitor(PKA)in group H89,dipyridamole plus H89 group,a total of five groups,each group simulation “during cerebral ischemia reperfusion”,then added with the appropriate drugs,analysis of endothelial cell F-actin(F-actin)distribution,tight junction protein expression of claudin 5,albumin and dextran permeability barrier,in order to determine the characteristics of various drugs on microvascular endothelial cells of rat brain function.Results Cilostazol and dipyridamole altered the distribution of endothelial fiber actin.Cilostazol increased the expression of tight junction protein claudin-5 by (218.2± 39.3)% compared to control with western blot quantitative assessment (P <0.01).Permeability to albumin was decreased to(0.869±0.15)×10 -6 cm/s in cilostazol group (P <0.05)and (0.65±0.11)×10-6 cm/s in dipyridamole group (P <0.01).Protein kinase A (PKA)inhibitors H89 attenuated the albumin permeability increased by cilostazol (P < 0.05).Permeability to dextran was decreased by cilostazol (P < 0.05 ),cilostazol + H89 (P < 0.05 ),and dipyridamole (P > 0.05 ). Conclusion These findings demonstrate distinctive effects of cilostazol and other PDE inhibitors on HBECs,including enhanced barrier characteristics and mitigation of response to histamine.PKA-mediated effects of cilostazol were prominent in this model.