中华血液学杂志
中華血液學雜誌
중화혈액학잡지
Chinese Journal of Hematology
2014年
6期
511-514
,共4页
王秀娟%张平平%赵丽丽%涂彧%戴克胜
王秀娟%張平平%趙麗麗%塗彧%戴剋勝
왕수연%장평평%조려려%도욱%대극성
活性氧%N-乙酰半胱氨酸%血小板%膜电位,线粒体%细胞凋亡
活性氧%N-乙酰半胱氨痠%血小闆%膜電位,線粒體%細胞凋亡
활성양%N-을선반광안산%혈소판%막전위,선립체%세포조망
Reactive oxygen species%N-acetylcysteine%Platelets%Membrane potential,mitochondrial%Apoptosis
目的 探讨活性氧在血小板凋亡中的调控作用.方法 健康人洗涤血小板用N-乙酰半胱氨酸(NAC)、辛可卡因和凝血酶单独或联合处理.用活性氧检测试剂盒结合流式细胞术检测血小板活性氧的产生和线粒体膜电位(△Ψm)的改变,用Western blot检测半胱氨酸天冬氨酸蛋白酶3(caspase-3)及抗凋亡蛋白Bcl-xL的表达情况.结果 ①NAC+辛可卡因组活性氧荧光值低于单用辛可卡因组(0.66±0.11对1.06±0.08,P=0.001),NAC+凝血酶组活性氧荧光值低于单用凝血酶组(0.45±0.05对0.71±0.11,P=0.001).②NAC+辛可卡因组线粒体膜电位正常的血小板百分数高于单用辛可卡因组[(86.30±9.37)％对(13.52±3.01)％,P＝0,000],NAC+凝血酶组高于单用凝血酶组[(93.00±3.03)％对(76.58±5.28)％,P＝0.000].③单用辛可卡因组caspase-3活化后的裂解片段明显多于DMSO对照组,NAC+辛可卡因组caspase-3活化后的裂解片段明显减少.④NAC+辛可卡因组Bcl-xL的表达较单用辛可卡因组明显增多,NAC+凝血酶组Bcl-xL的表达高于单用凝血酶组.结论 NAC对辛可卡因或凝血酶引起的血小板凋亡具有明显的抑制作用,这种抑制作用主要是通过对活性氧的调控而实现的.
目的 探討活性氧在血小闆凋亡中的調控作用.方法 健康人洗滌血小闆用N-乙酰半胱氨痠(NAC)、辛可卡因和凝血酶單獨或聯閤處理.用活性氧檢測試劑盒結閤流式細胞術檢測血小闆活性氧的產生和線粒體膜電位(△Ψm)的改變,用Western blot檢測半胱氨痠天鼕氨痠蛋白酶3(caspase-3)及抗凋亡蛋白Bcl-xL的錶達情況.結果 ①NAC+辛可卡因組活性氧熒光值低于單用辛可卡因組(0.66±0.11對1.06±0.08,P=0.001),NAC+凝血酶組活性氧熒光值低于單用凝血酶組(0.45±0.05對0.71±0.11,P=0.001).②NAC+辛可卡因組線粒體膜電位正常的血小闆百分數高于單用辛可卡因組[(86.30±9.37)％對(13.52±3.01)％,P＝0,000],NAC+凝血酶組高于單用凝血酶組[(93.00±3.03)％對(76.58±5.28)％,P＝0.000].③單用辛可卡因組caspase-3活化後的裂解片段明顯多于DMSO對照組,NAC+辛可卡因組caspase-3活化後的裂解片段明顯減少.④NAC+辛可卡因組Bcl-xL的錶達較單用辛可卡因組明顯增多,NAC+凝血酶組Bcl-xL的錶達高于單用凝血酶組.結論 NAC對辛可卡因或凝血酶引起的血小闆凋亡具有明顯的抑製作用,這種抑製作用主要是通過對活性氧的調控而實現的.
목적 탐토활성양재혈소판조망중적조공작용.방법 건강인세조혈소판용N-을선반광안산(NAC)、신가잡인화응혈매단독혹연합처리.용활성양검측시제합결합류식세포술검측혈소판활성양적산생화선립체막전위(△Ψm)적개변,용Western blot검측반광안산천동안산단백매3(caspase-3)급항조망단백Bcl-xL적표체정황.결과 ①NAC+신가잡인조활성양형광치저우단용신가잡인조(0.66±0.11대1.06±0.08,P=0.001),NAC+응혈매조활성양형광치저우단용응혈매조(0.45±0.05대0.71±0.11,P=0.001).②NAC+신가잡인조선립체막전위정상적혈소판백분수고우단용신가잡인조[(86.30±9.37)％대(13.52±3.01)％,P＝0,000],NAC+응혈매조고우단용응혈매조[(93.00±3.03)％대(76.58±5.28)％,P＝0.000].③단용신가잡인조caspase-3활화후적렬해편단명현다우DMSO대조조,NAC+신가잡인조caspase-3활화후적렬해편단명현감소.④NAC+신가잡인조Bcl-xL적표체교단용신가잡인조명현증다,NAC+응혈매조Bcl-xL적표체고우단용응혈매조.결론 NAC대신가잡인혹응혈매인기적혈소판조망구유명현적억제작용,저충억제작용주요시통과대활성양적조공이실현적.
Objective To study the effect of reactive oxygen species (ROS) on the regulation of platelet apoptosis.Methods Washed healthy human platelets were pre-incubated with N-caetyl-L-cysteine (NAC),and then stimulated with dibucaine or thrombin.The production of ROS and depolarization of mitochondrial membrane potential (△ Ψm) were detected by flow cytometry.The activation of caspase-3 and expression of Bcl-xL were analyzed by Western blot.Results ①The average ROS fluorescence value of NAC+dibucaine group was lower than that of dibucaine group (0.66±0.11 vs 1.06±0.08,P＜0.01),while that of NAC+thrombin group was also lower than that of thrombin group (0.45± 0.05 vs 0.71±0.11,P=0.001).②The percentage of platelets with normal △ Ψm in NAC+Dibucaine group was higher than that of dibucaine group [(86.30±9.37)％ vs (13.52±3.01)％,P=0.000],while that of NAC+thrombin group was also higher than that of thrombin group [(93.00±3.03)％ vs (76.58±5.28)％,P=0.000].③Fragmentation generated by caspase-3 activation in dibucaine group was much more than that in DMSO control group,while the fragmentation in NAC +dibucaine group was significantly decreased.④The expression of anti-apoptosis protein Bcl-xL of NAC+dibucaine group was significantly higher than that of the dibucaine group,while that of NAC + thrombin group was also higher than that of thrombin group.Conclusion Through the regulation of ROS,NAC could inhibit the platelet apoptosis induced by dibucaine or thrombin.
