摘要目的：探讨乙型肝炎病毒相关肝功能衰竭（HBV-LF）时循环组蛋白对肝细胞的损伤效应及其机制。方法收集 HBV-LF 患者血清，测定组蛋白 H3及 H4、ALT 、AST 、TBil 、DBil 、PTA 、肌酐和国际标准化比值（INR），计算患者终末期肝病模型（MELD）评分；比较患者与健康者血清中组蛋白水平，分析其与 PTA 、MELD 评分的相关性；用患者血清、人肝细胞裂解上清液和上清液＋抗组蛋白 H3、H4抗体分别处理培养的 L-02人肝细胞，观察细胞形态和凋亡率；用纯化组蛋白处理培养的 L-02细胞，观察细胞内钙离子浓度及半胱天冬氨酸蛋白酶（Caspase）-3活性。两独立样本均数比较采用 t 检验，相关性分析采用 Spearman 相关分析。结果与健康者比较，HBV-LF 患者循环组蛋白 H3[（5390．3±1032．0） mg/L 比（42．7±12．8） mg/L ，t ＝32．76，P＜0．01]、H4[（4205．1±662．3） mg/L 比（40．3±14．6） mg/L ，t＝39．74，P＜0．01]水平均显著升高；且患者循环组蛋白 H3、H4水平均与 PTA 呈负相关（r 值分别为-0．325、-0．572，均 P ＜0．05），与 MELD 评分均呈正相关（r 值分别为0．359、0．568，均P＜0．05）。含循环组蛋白 HBV-LF 患者血清或含组蛋白人肝细胞裂解上清液对培养的 L-02细胞均有损伤效应，该效应受抗组蛋白抗体抑制[（9．3±1．5）％比（14．3±0．6）％，t＝4．259，P＝0．02]；纯化组蛋白处理培养的 L-02细胞，细胞内钙离子荧光与 Caspase-3活性随组蛋白处理浓度增加而增强，抗组蛋白抗体对 Caspase-3活性增强效应有一定的抑制作用[（3．5±0．5）％比（5．2±0．6）％，t ＝4．243，P ＝0．02]。结论 HBV-LF 患者升高的循环组蛋白对肝细胞具有损伤作用。

abstractsObjective To investigate the injury effects and mechanisms of circulating histones on the hepatocytes in patients with hepatitis B virus (HBV )-related liver failure (HBV-LF) .Methods Serum samples from patients with HBV-LF were collected . The levels of serum histone H3 , histone H4 , prothrombin activity (PTA ) ,total bilirubin (TBil) ,creatinine (Cr) and international normalized ratio (INR) of the patients were measured .Model for end-stage liver disease (MELD) score was calculated in the patients .The serum levels of histones were compared between patients and the healthy volunteers . The correlation of histones with the MELD score and PTA was analyzed .The human liver cell line L-02 cells were cultured and treated with the serum of patients or L-02 cellular lysate supernatant preincubated with or without anti-histone H3 and H4 antibodies .The cellular morphology and rate of apoptosis were observed .Intracellular calcium ion concentration and Caspase-3 activity were detected in the cultured L-02 cells treated with histones .Mean of two independent samples was compared using t tests .Relationship between histones and the MELD score or PTA was conducted using Spearman correlation analysis .Results The levels of serum histones in the patients with HBV-LF were much higher than those in the healthy volunteers (H3 :[5 390 .3 ± 1 032 .0] μg/mL vs [42 .7 ± 12 .8] μg/mL , t = 32 .76 , P < 0 .01 ; H4 :[4 205 .1 ± 662 .3] μg/mL vs [40 .3 ± 14 .6] μg/mL ,t = 39 .74 , P< 0 .01) .In addition ,serum histones (H3/H4) levels in patients were negatively correlated with serum PTA (r= - 0 .325 ,P= 0 .038 and r =- 0 .572 ,P= 0 .028 ,respectively) ,but positively correlated with the MELD score (r= 0 .359 ,P= 0 .021 and r = 0 .568 , P = 0 .007 , respectively ) . Both serum of patients with HBV-LF and L-02 lysate supernatant were toxic to cultured L-02 cells .The injury effect was inhibited by anti-histone antibodies ([9 .3 ± 1 .5]% vs [14 .3 ± 0 .6]% , t = 4 .259 , P= 0 .02) .L-02 cells treated with calf thymus histone were cultured for 4 h . Cellular toxicity of histones resulted in Caspase-3 activation . The effect was inhibited by anti-histone antibodies ([3 .5 ± 0 .5]% vs [5 .2 ± 0 .6]% ,t= 4 .243 ,P= 0 .02) .Conclusion The elevated circulating histones in the patients with HBV-LF may aggravate the liver damage .