应用生物荧光法检测三磷酸腺苷评估肝细胞功能储备

背景：合并肝硬化肝病变患者接受大体积肝脏切除或其后进行肝移植手术可能造成较高的并发症发生率和死亡率。目前公认的肝功能评估方法是Child分级方法，包括了3个临床指标和2个生化指标，但它不能直接评估肝细胞的功能状态。目的：应用生物荧光法定量分析三磷酸腺苷作为肝功能监测手段的可行性。设计、时间及地点：病例对比观察，于2005—01／2006—03在中山大学附属第二医院完成。对象：选择肝部分切除患者32例，术中根据肝脏形态学改变分为3组：正常组7例，组织病理学没有明显硬化发现：大结节肝硬化组9例：微小结节型肝硬化组16例。方法：所有患者均于术前进行常规及肝生化检查，包括谷草转氨酶、总胆红素等。无菌收集肝手术切除病变旁肝组织标本，酶解消化、细胞悬浮、培养、离心、肝细胞计数，提取三磷酸腺苷并检测其含量。主要观察指标：换算出每个肘细胞的三磷酸腺苷含量，进步将三磷酸腺苷含量与术前肝功能指标进行相关性分析。结果：①大结节肝硬化组单位肝细胞三磷酸腺苷含量明显高于正常组及小结节肝硬化组（P=-0．0001，0．004）。小结节肝硬化组三磷酸腺苷含量也明显高于正常组（P=-0．04）。②单位肝细胞三磷酸腺苷含量与术前血浆谷草转氨酶活性、总胆红素含量呈显著正相关（r=-0．6093，P=0．0002：r=-0．6145，P=0．0002）。结论：肝细胞三磷酸腺苷水平检测可以评估肝实质储备功能，也可用于预测肝切除手术风险及选择肝移植手术时机。

Assessment of functional reserve of hepatocytes by bioluminescence adenosine triphosphate determination assay

BACKGROUND: Extensive liver resection or liver transplantation operated on patients with combined hepatic cirrhosis and other complications correlates with high morbidity and mortality.Child-Turcotte-Pugh scoring system is now widely used in the assessment of liver function.This classification scheme includes three clinical indicators and two biochemical indices;however,it seems difficulty on directly evaluating functional status of hepatocytes.OBJECTIVE: To explore the practicability of bioluminescence adenosine triphosphate (ATP) determination assay to assess the functional reserve of residual hepatocytes,DESIGN,TIME AND SETTING: Case contrast study,which was carried out in the Second Affiliated Hospital,Sun Yat-sen University from January 2005 to March 2006.PARTICIPANTS: Thirty-two patients who underwent major extra-and intra hepatic surgery including liver transplantation were randomly divided into three groups based on hepatic cirrhosis grading standard,including normal group (n=7),macronodular cirrhosis group (n=9),and micronodular cirrhosis group (n=16).METHODS: Routine examination and biochemical indexes of liver were performed preoperatively,including glutamic oxalacetic transaminase (GOT) and total bilirubin (TBIL).Liver specimens were delivered by aseptic technique during operation and enzymatic digested.Cell suspension was cultured and centrifuged.Hepatocytes were counted and dispensed cell suspension to be used for ATP extraction and measurement.MAIN OUTCOME MEASURES: ATP content,preoperative biochemical parameters of liver function,and correlation between biochemical parameters and ATP content.RESULTS: The ATP content in the macronodular cirrhosis group was significantly higher than that in the micronodular cirrhosis group and normal group (P=0.000 1,0.004).While,the ATP content in the micronodular cirrhosis group was also significantly higher than that in the normal group (P=0.004).ATP content (mole/cell) wassignificantly positively correlated with serum glutamic oxalacetic transarninase (r=-0.609 3,P=0.000 2) and TBIL (r=0.614 5,P=0.000 2).CONCLUSION: ATP assay can directly evaluate functional reserve of liver parenchyma and reflect high operative risk status (HORS) and course of postoperative recovery in major hepatic resection.