循环可溶性细胞色素c在肝病中作为细胞凋亡的标志物

Circulating soluble cytochrome c in liver disease as a marker of apoptosis.

作者信息

Ben-Ari Z, Schmilovotz-Weiss H, Belinki A, Pappo O, Sulkes J, Neuman M G, Kaganovsky E, Kfir B, Tur-Kaspa R, Klein T

机构信息

Liver Institute and Department of Medicine D, Beilinson Campus, Rabin Medical Center, Petah Tiqva, Israel.

出版信息

J Intern Med. 2003 Aug;254(2):168-75. doi: 10.1046/j.1365-2796.2003.01171.x.

DOI:10.1046/j.1365-2796.2003.01171.x

PMID:12859698

Abstract

OBJECTIVES

To measure levels of soluble cytochrome c, a clinical marker of apoptosis in patients with liver disease; determine whether soluble cytochrome c is derived from the liver; and correlate soluble cytochrome c level with histology and disease activity.

DESIGN

Laboratory research study with comparison group.

SETTING

Liver Institute, at the Rabin Medical Center, Israel, and In Vitro Toxicology Laboratory, Canada.

SUBJECTS

A total of 108 patients with liver disease and 30 healthy controls.

INTERVENTIONS

Paired hepatic and portal vein samples were taken via the transjugular vein in patients after liver biopsy and transjugular intrahepatic portacaval shunt, and bile from patients with external biliary drainage. Soluble cytochrome c was measured with an enzyme-linked immunosorbent assay in peripheral blood. Apoptotic cells in liver tissue were identified by morphological criteria and quantitated with the dUTP nick-end-labelling (TUNEL) assay.

MAIN OUTCOME MEASURES

Soluble cytochrome c level by type of liver disease by clinical and histological findings.

RESULTS

Soluble cytochrome c concentration (mean 187.1 +/- 219.5 ng x mL(-1)) was significantly higher in patients with liver disease than in controls (39.8 +/- 35.1 ng x mL(-1); P = 0.0001), with highest levels in the primary sclerosing cholangitis group (mean 1041.0 +/- 2844.8 ng x mL(-1); P = 0.001). Cytochrome c levels were correlated with serum bilirubin, alkaline phosphatase, creatinine levels, necroinflammatory score and apoptotic index, but not with serum alanine aminotransferase and synthetic liver function tests. In the 16 paired samples, soluble cytochrome c level was higher in the hepatic (mean 267.9 +/- 297.0 ng x mL(-1)) than the portal vein (mean 169.2 +/- 143.3 ng x mL(-1)), and it was highly detectable in bile (mean 2288.0 +/-4596.0 ng x mL(-1)) (P = 0.001). Untreated patients with chronic viral hepatitis (B and C) had significantly higher levels (mean 282.8 +/-304.3 ng x mL(-1)) than treated patients (77.9 +/- 35.8 ng x mL(-1); P = 0.001).

CONCLUSIONS

Soluble cytochrome c levels are increased in different types of liver disease. Soluble cytochrome c is probably derived from the liver and secreted into the bile. Levels correlate with the apoptotic index and are affected by antiviral treatment. Soluble cytochrome c may serve as a serum marker of apoptosis.

摘要

目的

测量可溶性细胞色素c水平，其为肝病患者细胞凋亡的临床标志物；确定可溶性细胞色素c是否来源于肝脏；并将可溶性细胞色素c水平与组织学及疾病活动度相关联。

设计

设有对照组的实验室研究。

地点

以色列拉宾医疗中心肝病研究所及加拿大体外毒理学实验室。

研究对象

共108例肝病患者及30名健康对照者。

干预措施

在肝活检及经颈静脉肝内门体分流术后，通过经颈静脉采集患者配对的肝静脉和门静脉样本，以及外引流胆汁患者的胆汁样本。采用酶联免疫吸附测定法检测外周血中的可溶性细胞色素c。通过形态学标准鉴定肝组织中的凋亡细胞，并用脱氧尿苷三磷酸缺口末端标记法（TUNEL法）进行定量分析。

主要观察指标

根据临床及组织学检查结果，按肝病类型划分的可溶性细胞色素c水平。

结果

肝病患者可溶性细胞色素c浓度（均值187.1±219.5 ng·mL⁻¹）显著高于对照组（39.8±35.1 ng·mL⁻¹；P = 0.0001），其中原发性硬化性胆管炎组水平最高（均值1041.0±2844.8 ng·mL⁻¹；P = 0.001）。细胞色素c水平与血清胆红素、碱性磷酸酶、肌酐水平、坏死性炎症评分及凋亡指数相关，但与血清丙氨酸氨基转移酶及肝脏合成功能试验无关。在16对配对样本中，肝静脉中可溶性细胞色素c水平（均值267.9±297.0 ng·mL⁻¹）高于门静脉（均值169.2±143.3 ng·mL⁻¹），且在胆汁中高度可检测到（均值2288.0±4596.0 ng·mL⁻¹）（P = 0.001）。未经治疗的慢性病毒性肝炎（B型和C型）患者水平（均值282.8±304.3 ng·mL⁻¹）显著高于接受治疗的患者（77.9±35.8 ng·mL⁻¹；P = 0.001）。