肝星状细胞连接对大鼠肝纤维化病理过程的影响

发布时间：2018-04-26 12:35

本文选题：肝纤维化 + 肝星状细胞　；参考：《大连医科大学》2016年硕士论文

【摘要】：背景:肝纤维化(Liver Fibrosis)指肝脏内弥漫性细胞外基质过度沉积的病理过程,是各种慢性肝病共同的病理基础,目前关于肝纤维化机制的研究已经取得了很大的进展。肝纤维化形成的中心环节为肝星状细胞的活化,当肝脏受到酒精、自身免疫、肝炎病毒等因素刺激时,静止的肝星状细胞会转化为具有增生性、收缩性,可表达α-平滑肌肌动蛋白(alpha Smooth Muscle Actin,α-SMA)的肌成纤维细胞,从而生成细胞外基质,最终导致肝纤维化的产生。目前关于肝星状细胞活化的研究主要集中在细胞因子上,肝纤维化的治疗也通过抑制细胞因子而发挥作用,但始终无法达到预期的疗效,因此可能存在着细胞因子以外的促活化机制。目前四氯化碳诱导的大鼠肝纤维化模型在病理形态及特征上与人类肝纤维极其相似,因此可通过此模型探究肝星状细胞的活化机制。目的:通过四氯化碳诱导的大鼠肝纤维化模型,观察肝星状细胞连接对肝纤维化病理过程的影响。方法:28只SD大鼠随机分为肝纤维模型组(n=24)和对照组(n=4)。肝纤维化模型组应用40%四氯化碳橄榄油混合溶液按照1.0ml/Kg体重对SD大鼠进行灌胃,2次/周,持续12周建立肝纤维化模型;对照组以同一时间与肝纤维化模型组等量的蒸馏水灌胃,两组大鼠均给予相同基础饮食。利用HE染色观察大鼠肝组织炎症浸润程度,Masson染色观察大鼠肝组织纤维化程度。免疫组织化学荧光染色观察肝星状细胞结蛋白(Desmin)和α-平滑肌肌动蛋白(α-SMA)的表达。免疫组织化学荧光双重染色观察活化肝星状细胞的形态及分布,总结肝星状细胞活化规律,仔细观察活化肝星状细胞的排列方式与纤维化形成的关系。结果:1.正常肝组织(G0S0)内无活化的肝星状细胞,静止肝星状细胞均匀分布于肝窦内。2.在肝纤维化形成前的炎症阶段(G1S0),肝窦内的肝星状细胞受到炎症刺激后开始活化,此时肝星状细胞仍均匀分布在肝窦内。3.肝纤维化形成早期(G2S1及G3S2),肝星状细胞活化增强并迁移到炎症区域,无炎症区域几乎无肝星状细胞。4.纤维间隔形成阶段(S4期),肝星状细胞较早期明显活化,胞体增大,肝星状细胞分布并非简单的聚集于纤维化区域,而是呈明显的“首尾相接”方式,展现了细胞“链接”的模式。结论:四氯化碳诱导SD大鼠肝纤维化模型中肝星状细胞活化是导致肝纤维化的重要因素,细胞连接是纤维间隔形成的重要条件。
[Abstract]:Background: liver fibrosis refers to the pathological process of excessive deposition of diffuse extracellular matrix in the liver, which is the common pathological basis of various chronic liver diseases. At present, great progress has been made in the study of the mechanism of liver fibrosis. The central link in the formation of hepatic fibrosis is the activation of hepatic stellate cells. When the liver is stimulated by alcohol, autoimmunity, hepatitis virus and other factors, the stationary hepatic stellate cells will be transformed into proliferative and contractile cells. Myofibroblasts expressing 伪 -smooth muscle actin alpha Smooth Muscle actin (伪 -SMAA) produce extracellular matrix (ECM), leading to liver fibrosis. At present, researches on the activation of hepatic stellate cells mainly focus on cytokines. The treatment of liver fibrosis also plays a role by inhibiting cytokines, but it can not achieve the expected curative effect. Therefore, there may be a pro-activation mechanism beyond cytokines. The present rat liver fibrosis model induced by carbon tetrachloride is very similar to that of human hepatic fiber in pathological morphology and characteristics, so the activation mechanism of hepatic stellate cells can be explored by this model. Aim: to observe the effect of hepatic stellate cell junction on the pathological process of hepatic fibrosis in rats induced by carbon tetrachloride. Methods 28 Sprague-Dawley rats were randomly divided into liver fiber model group and control group. Rats in hepatic fibrosis model group were treated with 40% carbon tetrachloride olive oil mixed solution twice a week for 12 Zhou Jianli according to 1.0ml/Kg body weight, and the control group were fed with distilled water at the same time as the liver fibrosis model group, and the control group was given the same amount of distilled water at the same time as the liver fibrosis model group. Both groups were given the same basic diet. He staining was used to observe the degree of inflammatory infiltration in rat liver tissue and Masson staining was used to observe the degree of hepatic fibrosis in rats. The expression of desmin and 伪-smooth muscle actin (伪 -SMA) in hepatic stellate cells was observed by immunohistochemical fluorescence staining. The morphology and distribution of activated hepatic stellate cells were observed by immunohistochemical double staining. The relationship between the arrangement of activated hepatic stellate cells and the formation of fibrosis was carefully observed. The result is 1: 1. There were no activated hepatic stellate cells in normal liver tissue (G0S0), and stationary hepatic stellate cells distributed uniformly in sinusoidal hepatic sinuses. During the inflammatory stage before hepatic fibrosis, hepatic stellate cells in hepatic sinusoids were activated after being stimulated by inflammation, and hepatic stellate cells were still distributed in hepatic sinusoids. In the early stage of hepatic fibrosis, the activation of hepatic stellate cells was enhanced and migrated to the inflammatory region, and almost no hepatic stellate cells were found in the non-inflammatory areas. In the S4 phase of fibrous septum, hepatic stellate cells were obviously activated and their cell bodies were enlarged. The distribution of hepatic stellate cells was not simply concentrated in the fibrotic region, but in a "head-and-tail" pattern. Shows the cell "link" pattern. Conclusion: the activation of hepatic stellate cells is an important factor in hepatic fibrosis induced by carbon tetrachloride in SD rats.
【学位授予单位】：大连医科大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R575.2

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