【作者】 杨日高；

【导师】 王曙光；

【作者基本信息】 第三军医大学 ， 外科学， 2002， 硕士

【摘要】 目的：肝脏是一个具有多种生理机能的器官，并且有巨大的再生潜能和代偿能力，它参与机体的消化、代谢、解毒、排泄和免疫等多种过程，如何准确评估肝脏储备功能是目前肝病领域的重要课题之一。 传统肝储备功能的定量评估方法很多。有肝功能Child分级法、Child—Pugh分级方法、佐藤分级法等。ICG负荷试验是目前常用的评估肝脏储备功能的方法之一。以99mTc标记去唾液酸糖蛋白(GSA)作为特异性配体，通过SPECT闪烁扫描测定肝脏去唾液酸糖蛋白(ASGP)受体的量，以此评估肝脏储备功能是近年来开展的定量评估肝脏储备功能的新方法。文献报道该方法能较直观地反映肝脏清除去唾液酸糖蛋白的能力。相映指标可以定量评估肝脏储备功能。用流式细胞仪对分离的肝细胞进行ASGP受体分析以评估肝脏储备功能是处于实验阶段的新方法。 去唾液酸胎球蛋白(ASF)是肝细胞ASGP受体的天然配体，能被ASGP受体特异结合并被肝细胞清除，被广泛用于肝脏ASGPR研究。 能否将吲哚氰绿负荷试验和肝脏ASGP受体核素扫描显像相关指标进行有机的结合，探索一种更加准确更加敏感的对肝脏储备功能进行定量评估的方法? 针对以上问题，本研究试图以99mTc—ASF作为配体探讨SPECT肝脏ASGP受体扫描的有关指标与常规肝功能指标间的相关性，并比较各项指标的灵敏性。选择其中比较敏感的指标作参数，建立一个综合定量评估肝脏储备功能的评分体系，为临床提供参考。 方法：以雄性Wistar大鼠为试验对象，建立四氯化碳肝硬化模型、肝脏不同比例部分切除模型，分别进行常规肝功能指标测定、吲哚氰绿储留率测定、肝脏ASGP受体核素扫描测定15min配体清除率(HH15)、受体指数（LHL15）、受体修正指数（MRI）、肝细胞 ASGP受体体外流式细胞仪（FCM）分析等，探讨各项指标的相关性及敏感性。对不同肝损害肝切除模型进行常规肝功能指标、ICGR15以及mI、HH15、LHL15等指标的综合测定，通过回归分析，寻求一种涵盖多项指标的综合评估肝脏储备功能的方法。 统计学方法：统计资料表达采用M土SD。统计学处理采用SPSS及Microsoft Excel软件进行单因素方差分析、COX回归、线性相关分析、计数资料用X’检验。确定P狈．05为有统计学意义。 结果：在四氯化碳肝硬化模型及肝切除模型中，反映肝脏功能状态方面的指标之间多数有显著相关性；FCM分析正常大鼠肝细胞的ASGP受体量明显高于肝硬化大鼠的该受体量。在肝脏容量丢失15％以内，ICGR15不能反映出肝脏储备功能的变化，即该指标不够敏感。相关指标的敏感性依次为：MRI、HH15汀CGR15儿HL15。通过回归分析建立的综合评估肝脏储备功能的回归方程式：Y＝33．188 X MRI—44．844 XHH；s＋24．032XLHL；s一34．915 X ICGR；s能比较敏感地反映肝脏储备功能。当评估系数 Y值小干 30时，大鼠在72小时内100％死亡；当Y值在30～40之间时，72小时内死亡率为37．46％；当Y值40以上时，大鼠在72 ’J’时内没有死亡。以Y＝＝35为界，当Y低于该值时大鼠72小时内的死亡率为81．8％。 结论：1、在大鼠四氯化碳肝硬化模型及肝脏部分切除模型中，常用评估肝脏储备功能指标之间的相关性多数有显著的统计学意义。2、用流式细胞仪对肝细胞ASGP受体进行体外分析能够反映肝脏细胞ASGP受体量，可以反映肝脏的功能状态。3、涵盖MRI、HH15、ICGR15、LHL15等四项指标的定量评估肝脏储备功能的回归方程 Y ＝＝ 33．188 X Mi一 44．844 XHH15＋24刀32 XLHL15—34．gl5XICGR15，能够对正常及四氯化碳肝硬化大鼠模型的肝脏储备功能作出准确可靠的定量评估。更多还原

【Abstract】 Objective: Liver is an organ that posses multiple physiological function, great regeneration potential and compensation. A great deal of processes such as digestion, metabolism, detoxication, excretion and immunity are undertaken in liver. At present, how to evaluate hepatic function reserve accurately is a challenge in treatment of liver disease.There are many classical methods evaluating hepatic function reserve quantitatively, including Child classifying method of hepatic function, Child-Pugh classifying method, Safuji classifying method, and so on. ICG stress test is the popular method evaluating hepatic function reserve. In recent years, the quantitation of ASGP receptor by 99mTc-labeling neogalactoalbumin SPECT scintiscan is the up-to-date method that is applied to quantitatively evaluate hepatic function reserve. Literature reported that the capability of eliminating ASGP of the liver can be reflected more directly by this method, and correlative index can be applied to quantitatively evaluate hepatic function reserve. Analysing the surface ASGP receptor on separated liver cell by flow cytometer, and is a new method evaluating hepatic function reserve, however, this kind of study is still at its experiment stage. Asialofetuin (ASF) is the ligand of ASGP receptor residing on liver cell surface, can bind its receptor specifically and be degradateded by liver cell. Now ASF is applied to investigate ASGP receptor of liver extensively.Whether or not can the indocyanogen green retention rate test be combined effectively with the correlative index of liver ASGP receptor nuclin scanning, so that a more accurate and more sensitive method of evaluatingquantitatively hepatic function reserve can be selected.Aiming at above problems, the objective of this experiment is to discuss correlation between indices of SPECT liver ASGP receptor scintiscan with the ligand of 99mTc - ASF and classical liver functional index, and sensitivity of those indices. Further, we hope to choose a more sensitive parameter from those indices, and provide a guiding standard for clinical staff to select a quantitatively evaluation method of hepatic function reserve.Methods: Wistar rats (male) were used to establish a model of carbon tetrachloride inducing cirrhosis and differently proportional resection of rodent liver. Classical liver functional index, indocyanogen green retention rate test, 15min ligand clearance estimated by liver ASGP receptor nuclin scanning (HH15), liver uptaking rate (LHL15), modified receptor index (MRI), and in vitro analysis of ASGP receptor on liver cell surface detected by Flow cytometer were investigated respectively. The correlation and sensitivity of each index were discussed.The indices of differently damaged liver model and differently proportional resection of rodent liver such as Classical liver functional index, ICGR15,MRI,HH15,and LHL15,were be evaluated combined. And then a synthetic method including many indices of evaluating hepatic function reserve could be found out by regression analysis.Statistic methods: Statistic data were expressed as M SD. And those data were analysed by software of SPSS and Microsoft Excel. Single equation variance analysis, COX regression, linearity correlation analysis , and X2 test were undertaken. The test standard was p<0.05.Results: In the models of carbon tetrachloride inducing cirrhosis and differently proportional resection of rat liver, there was a significant correlation among commonly used indexes of hepatic functional reserve. The quantity of ASGP receptor of normal liver were significantly more than that of cihhorsisby FCM. In the 15% losing of hepatic dimension, the change of hepatic functional reserve can not be reflected by ICGR15,that means this index is not sensitive enough. The sensitivity of those indices was that MRI , HH15>ICGR15>LHL15.The regression equation of evaluating hepatic functional reserve was that Y= 33.188 MRI-44.844 HH15+24.032 LHL15 - 34.915 ICGR15, and it was sensitive. When the evaluation coeffic更多还原