title: [Study of reactive oxygen species on the regulation of platelet apoptosis].
abstract: 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-Lcysteine (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
  (1)The 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). (2)The 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]. (3)Fragmentation 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. (4)The 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.
  
  
  CONCLUSIONS
  Through the regulation of ROS, NAC could inhibit the platelet apoptosis induced by dibucaine or thrombin.
authors.affiliation: Jiangsu Institute of Hematology, the First Affiliated Hospital of Soochow University, Key Laboratory of Thrombosis and Hemostasis, Ministry of Health, Suzhou 215006, China.
authors.forename: Xiujuan
authors.initials: X
authors.lastname: Wang
authors.affiliation: Jiangsu Institute of Hematology, the First Affiliated Hospital of Soochow University, Key Laboratory of Thrombosis and Hemostasis, Ministry of Health, Suzhou 215006, China.
authors.forename: Pingping
authors.initials: P
authors.lastname: Zhang
authors.affiliation: Jiangsu Institute of Hematology, the First Affiliated Hospital of Soochow University, Key Laboratory of Thrombosis and Hemostasis, Ministry of Health, Suzhou 215006, China.
authors.forename: Lili
authors.initials: L
authors.lastname: Zhao
authors.affiliation: Jiangsu Institute of Hematology, the First Affiliated Hospital of Soochow University, Key Laboratory of Thrombosis and Hemostasis, Ministry of Health, Suzhou 215006, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Tu
authors.affiliation: Jiangsu Institute of Hematology, the First Affiliated Hospital of Soochow University, Key Laboratory of Thrombosis and Hemostasis, Ministry of Health, Suzhou 215006, China.
authors.forename: Kesheng
authors.initials: K
authors.lastname: Dai
mesh_terms: D000111:Acetylcysteine / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D001792:Blood Platelets / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D053148:Caspase 3 / Q000378:metabolism; D003992:Dibucaine / Q000494:pharmacology; D006801:Humans; D053078:Membrane Potential, Mitochondrial / Q000502:physiology; D017382:Reactive Oxygen Species / Q000378:metabolism*; D013917:Thrombin / Q000494:pharmacology; D051020:bcl-X Protein / Q000378:metabolism
chemical_list: C494811:BCL2L1 protein, human; D017382:Reactive Oxygen Species; D051020:bcl-X Protein; D013917:Thrombin; C505407:CASP3 protein, human; D053148:Caspase 3; D003992:Dibucaine; D000111:Acetylcysteine