体外高迁移率族蛋白1对人循环中性粒细胞凋亡的影响

Effects of HMGB1 in vitro on human circulating PMN apoptosis

目的观察体外高迁移率族蛋白1(HMGB1)对人循环PMN凋亡的影响,以探讨HMGB1在ALI的发病机制中的作用。方法分离提纯人静脉血PMN,与不同浓度的HMGB1在37℃共孵育24h后,采用流式细胞术(FCM)及TUNEL法检测其凋亡情况。结果流式细胞术分析PMN的凋亡率分别为(40.53±4.12)%、(40.52±2.73)%、(34.89±1.15)%及(18.77±3.02)%,与阳性、阴性对照组比较有统计学意义;PMN TUNEL阳性率分别为(31.42±4.40)%、(31.39±3.80)%、(25.62±2.46)%及(17.98±3.20)%,与对照组比较,呈HMGB1剂量依赖性降低。结论体外HMGB1可明显抑制人循环PMN细胞凋亡,该抑制效应呈HMGB1剂量依赖性。HMGB1在脂多糖诱导的急性肺损伤(ALI)PMN凋亡改变中有重要的调节作用。

Objective To observe effects of HMGB1 in vitro on human circulating PMN apoptosis, therefore to clarify the role of HMGB1 in the pathogenesis of acute lung injury. Methods Separated and purified human circulating PMN was coincubated for 24h with different doses of HMGB1 （0, 10, 100,1000 ng/ml, respectively） at 37℃ in 5% CO2. PMN apoptosis rate was determined by flow cytometric （FCM） analysis and by TdT-mediated dUTP nick end labeling （TUNEL） method. Results By flow cytometry, PMN apoptosis ratio was （40.53± 4.12） %, （40. 524± 2.73） %, （34.89± 1.15） % and （18. 77±3.02）%, respectively. There was statistic significance. Meanwhile, PMN TUNEL positive rate was （31. 42± 4.40） %, （31.39± 3.80） %, （25.62 ± 2.46）% and （17.98± 3.20）%, respectively. The inhibitory effect was HMGB1 dose-depended as compared with that of control group. Conclusion HMGB1 in vitro inhibited human circulating PMN apoptosis markedly, and the inhibitory effect was HMGB1 dose-depended. HMGB1 may play an important role as a modulator in apoptotic changes in PMN during LPS-induced ALL.