目的:以CCl_4大鼠肝纤维化为模型,研究肝与他脏在生理、病理之间的相互联系,为中医理论对“以象测脏”认识脏象的方法论提供科学依据,探讨中医整体观念的物质基础。方法:将 Wistar大鼠,随机分正常组、模型组。模型大鼠皮下注谢40％CCl_4-橄榄油溶液每周2次,共2周; 停止剌激后再观察4周,正常组以生理盐水替代CCl_4皮下注射。分别于第4、8、12、16周末分批处死大鼠。留取肝组织分别做病理、羟脯氨酸、蛋白质的分步提取。蛋白质定量后进行二维电泳,凝胶银染,运用PDQUEST 2-DE图象分析软件对获得的蛋白质图谱进行分析,运用MALDI-TOF-MS 鉴定了30多个差异表达的蛋白质。结果:①CCl_4肝纤维化大鼠出现生命活动整体反应,如体重、活动度等下降;②肝纤维化过程中存在蛋白质合成和物质代谢碍障;③蛋白质组改变的质谱鉴定。结果显示:肝纤维化大鼠与正常大鼠肝组织差异表达的蛋白质与物质代谢相关的蛋白质有过氯酸可溶性蛋白质、磷脂酰肌醇转移酶、磷酸甘油酸激酶;与神经内分泌相关的蛋白质分子有儿茶酚-邻 -甲基转移酶、雌激素转磺酶亚型6、转录起始因子、非粘附性细胞蛋白质。结论:肝与他脏功能相关、病理相连是以蛋白质分子为物质基础,肝纤维化是一个涉及多个功能的整体病变过程,而引起机体整体功能改变的物质基础是蛋白质分子。

To investigate the relationship between liver and other vicera in physiology and pathology by taking the rats with liver fibrosis CC14 for models so as to provide scientific basis for understanding the methodology concerning visceral state in the TCM theory of "testing vescera via the state of internal organs" and explore the material foundation of the concept of wholism in TCM. Method Rats Wistar are divided into two groups, normal and model at random. The rats in model group are subcutaneously injected with 40% CC14 -carbon tetrachloride -olive solution twice a week in successive 12 weeks and are observed for 4 weeks after stopping stimulation of them while the rats in normal group are injected with physiological saline instead of CC14. All the rats of the two groups are killed at the end of 4、8、12 and 18 weeks respectively and the pathological observation, hydroxyproline determination and proteome preparation of their liver tissues are conducted step by step. 2 - dimensional electrophoresis and silver staining are made after the quantitativeness of protein and the spectra of protein obtained is analyzed by the 2 - DE software for image analysis. More than 30 proteins with differential expression have been identified by the method of MALD1-TOF- MS. Result （1） Rats with liver fibrosis due to the injection of CCI4 assume the general reflection of their biological activities, such as the decrease in weigh and activity; （2） There exists disturbance of protein synthesis and the metabolism of matters in the process of liver fibrosis; （3） The result of mass spectra due to the change of proteome shows that proteins related to the metabolism of matters, such as perchloric acid soluble protein, phosphatidylinositol transfer protein, phoshoglycerate kinase and ER - 60 protease and protein molecules related to neuroendocrine, such as catechol -O-methyltransferase, estrogen sulfotransferase isoform 6, transcription initiation factor TFIID 28 KD2 subunit and NAC - 1 protein are indentified in the differentially expressed proteins of the liver tissues of the rats with liver fibrosis and the normal ones. Conclusion Protein molecules constitute the material foundation on which liver links to the functions and pathology of other viscera. Liver fibrosis represents a process of generally pathological change which refers to multiple functions. The material foundation which brings about the change of the whole functions of organs, however, is protein molecules.

国家自然科学基金重大计划重点项目(90409020):中医方剂的方证症态生物信息模式的探索,负责人：刘平；国家自然科学基金项目(30371835)：扶正化瘀干预大鼠肝纤维化肝组织蛋白质组动态变化的研究，负责人：刘莺。