摘要
目的阐明炎性过程中氧化和抗氧化水平对脐血粒细胞生长和凋亡的调节作用。方法体外肝素化脐血加脂多糖(LPS)模拟内毒素血症,设①对照组;②实验组:加入1μg/ml LPS;③治疗组:加入LPS 1μg/ml前加N-乙酰半胱氨酸(NAC)10 mmol/L孵育30 min。采用luminol增强化学发光检测活性氧(ROS),放射性免疫技术检测肿瘤坏死因子(TNF-α),Western blot检测粒细胞胞质和胞核中核因子(NF-κB)的表达,采用粒细胞集落(CFU-G)形成评价粒细胞的生长和流式细胞术检测粒细胞的凋亡和坏死。结果①实验组ROS产生水平高于对照组和治疗组(P<0.05)。②实验组LPS作用2、6、122、4 h时的TNF-α水平明显高于对照组(P<0.05),在2、6 h时明显高于治疗组(P<0.05)。③LPS刺激激活NF-κB p65表达从胞质转移到胞核,NAC治疗抑制了NF-κB的激活。④实验组CFU-G数目明显低于对照组和治疗组(P<0.05)。⑤脐血粒细胞体外培养24 h,实验组与对照组相比凋亡率明显降低(P<0.05),坏死率明显增高(P<0.01);治疗组与实验组相比凋亡率升高,坏死率降低(P<0.05)。结论抗氧化剂通过清除LPS刺激粒细胞产生的ROS,可调节粒细胞的生长和凋亡。
Objective To study the oxidant and antioxidant modulatory effects on umbilical cord blood granulopoiesis and granulocyte apoptosis in response to lipopolysaccharide （LPS） treatment and to apply antioxidant N-acetyl-cysteine （NAC） to regulate inflammatory processes. Methods LPS was added to samplcsof beparinized umbilicalcord blood in vitro to mimic endotoxemia. Following groups were set up： normalcontrol group, experiment group （LPS 1 μg/ml）. treatment group （NAC 10 mmol/L added before LPS 1 μg/ml was given）. Endotoxin elicited reactive oxygen species （ROS） was determined by chemiluminescence and tumor necrosis factor alpha （TNF-α） by radioimmunoassay, respectively. Nuclear factor-kappaB （NF-κB） of nuclear and cytoplasm was detected by Western blouing. Growth of granulocyte was assessed by colony-forming unh-granulocyte formation （CFU-G）. Apoptosis and necrosis of granulocytes were detected by flow cytometry. Results ①ROS was induced in response to LPS. The level of ROS in experiment group was higher than in normal control group and treatment group （both P〈0. 05）. ② The levels of TNF-α at 2. G. 12 and 24 h after LPS stimulation were significantly increased in experiment group and treatment group as compared with those in normal control group （P〈0.05）. ③Stimulation with LPS induced NFκB p65 of granulocytes translocated from the cytoplasm to the nucleus. NAC could inhibit the activation of NF-κB. ④ The amount of CFU G in experiment group was significantly less than in normal control group and treatment group （both P〈0.05）. ⑤ Apoptotic rate was significantly reduced （P〈0. 05） and necrotic rate increased （P〈0.01） in experiment group as compared with normal control group, while in treatment group, the apoptotic rate was higher and necrotic rate lower than in normal control group （both P〈0. 05 ）. Conclusion ROS produced by immanc cells plays an important part during the process of inflam,, marion. Oxidant and antioxidant have differe
出处
《华中科技大学学报（医学版）》
CAS
CSCD
北大核心
2006年第1期90-92,114,共4页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
关键词
内毒素
活性氧
粒细胞
细胞凋亡
抗氧化作用
endotoxin
reactive oxygen spcies
granulocyte
aoptosis
antioxdation
