| IntroductionThe perivenous fibrosis along the sinusoid space is one of the pathological characters of the alcoholic liver fibrosis. It is mostly regarded that this is associated with many cells, such as hepatocyte, hepatic stellate cell ( HSC) , kupffer cell ( KC) , liver sinusoid epithelial cell (LSEC) and myofibroblast, as well as many cytokines, for example, epithelial growth factor (EGF) , interleucin -2 (IL-2) and interleucin -10 (IL-10), tumor necrosis factor (TNF) and platelet derived growth factor (PDGF).LSEC can be regarded as unique capillary endothelial cell with fenestrae lacking a diaphragm and a basement membrane ( BM ) . It is the first barrier of the liver, and many various substances first effect on the LSEC brought from portal venous and absorbed from intestinal gut. The long continuous effect on it can make fenestrae gradually decrease or disappear ( defenestrae) , BM gradually form and even develop to sinusoid capillarization, which is the most basal character of the earlier liver fibrosis.The abuse of chronic ethanol can induce alcoholic steatosis, fatty necrosis (hepatics) , liver fibrosis and cirrhosis, and there is little knowledge about the relation between pathological changes above and the LSEC. Some studies regarded that a little amount of ethanol drinking in a short time could dilate the fenestrae and promote the fat metabolism, whereas the chronic ethanol could induce a decreased number of fenestrae and polarity, and induce the hyperlipoproteinmia and atherosclerosis related to ethanol toxin. It can be seen that ethanol has a critical effect on the LSEC morphology and structure.There are (different definitions on the amount and duration of ethanol affected the LSEC by various scholars, and different animals have different structureof LSEC, too. Moreover, there is little continuous observation on the study.In the previous animal studies, we not only successfully established the animal model of rat liver alcoholic fibrosis, but also systemically and continuously observed the changes of liver pathology and many cytokines, HSC effects, extra cellular substance formation and relative factor degrading, and so on, during the development of liver fibrosis induced by ethanol. We also observed that Kang Xian Fu Fang I ( KXI) can significandy inhibit HSC secretion, MMP - 2, - 9 expression, TGF - beta 1 and PDGF secretion caused by acetaldehyde and can also affect the collagen formation, thus can prevent and even treat the alcoholic liver disease (ADD ) .Based on the previous studies, we further continuously observed in this paper the changes of the rat LSEC morphology and structure induced by ethanol in vivo and in vitro, and continuously interfered the LSEC with KXI, therefore, further explored the theological basis of the effects of ethanol on the LSEC morphology and structure and the preventing function of KXI on the ALD.Material and Method1. Experimental material and equipment1.1 Experimental animalsMale Wistar rats (200g 10g) , provided by animal department of China Medical University.1.2. Experimental solutionsHeat - inactivated fetal calf serum ( FCS - HI, obtained from Hycolon company ) , Collagenase A and collagen - S ( from Roche company) , DNase I (from Hua Mei company) , RPMI - 1640 ( from Gibco campariy) , Endothelial cell growth factor (from Roche company).KXI (mainly Include safflower, angelica, astragalus membranaceus, saliva miltiorrhiza, tetrandrine and so on).1. 3 Experimental equipmentBeckman centrifuge, Philip scan electron microscopy ( SEM ) EM400, JEM - 1200EX transmission electron microscopy ( TEM) , fluorescence microscope(328nm excitation light) , Olympus light microscope.2. Method of establishing animal model2. 1 The establishment of ALD experimental animal model ( method of ethanol oral - gastric administration) .Reference to the established model method by our experimental department (reference 33 )2. 2 The establishment of KXI preventing experimental animal model. Reference to the es...
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