【作者】 李响；

【导师】 魏伟；

【作者基本信息】 安徽医科大学 ， 药理学， 2009， 硕士

【摘要】 肝脏接受门静脉血流,是机体面对酒精、药物毒物、病毒细菌等损伤因素的重要代谢器官。肝脏免疫细胞的组成数量和活化状态与其他免疫组织有着较大的区别,当药物过量(对乙酰氨基酚)、酒精中毒、器官移植(缺血再灌注)、暴发型肝炎、大面积创伤和内毒素休克,往往会引起严重的损伤和炎症反应,在急性期炎症级联反应中,容易诱发严重的弥漫性血管内凝血和多器官功能衰竭而导致死亡。在这些疾病中肝炎是大面积疾患的重要原因,病理表现为肝细胞变性、凋亡、坏死,是多种肝脏疾病共同的起始因素,慢性迁延的损伤使得肝脏逐渐丧失正常的组织结构,逐渐发展成为肝硬化、肝癌。因此,及时的积极的针对多种损伤因素导致的肝脏疾病进行控制将对临床有着重要的意义。目的:探讨芍药苷(paeoniflorin, Pae)对卡介苗(Bacillus Calmette-Guérin, BCG)加脂多糖(lipopolysaccharide, LPS)诱导的小鼠免疫性肝损伤的作用及其部分机制。方法:建立BCG+LPS诱导小鼠免疫性肝损伤模型,分光光度法检测血清丙氨酸氨基转移酶(alanine aminotransferase,ALT)、天门冬氨酸氨基转移酶(aspartate aminotransferase,AST)、一氧化氮(nitric oxide,NO)水平和肝组织中丙二醛(malondiadehyde,MDA)、超氧化物歧化酶(superoxide dismutase,SOD)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)的含量;建立大鼠免疫性肝损伤模型,分光光度法检测血清ALT、AST,肝脏原位灌流分离枯否细胞(kupffer cell,KC)、肝星状细胞(Hepatic Stellate Cellular, HSC),进行共培养,观察KC对HSC增殖的作用,MTT法检测KC培养上清液对HSC增殖的影响和Pae的作用。HE染色法对肝脏组织作病理检查。结果:(1)Pae(53、105、210 mg·kg-1)和TGP(85 mg·kg-1)灌胃给药能减少免疫性肝损伤小鼠死亡;(2)Pae(53、105、210 mg·kg-1)和TGP(85 mg·kg-1)能降低模型组升高的肝脾指数、降低过高的血清转氨酶水平、减少炎性细胞浸润、减轻肝组织坏死程度和范围、降低肝组织匀浆中升高的MDA水平以及升高SOD、GSH-Px酶活性。(3)Pae 10-7~10-5 mol·L-1作用后能抑制KC培养上清促HSC的增殖的作用,免疫性肝损伤大鼠KC与HSC共培养发现,KC能明显促进HSC的增殖。结论:Pae对小鼠免疫性肝损伤具有减轻作用;对肝损伤大鼠体外KC促HSC增殖具有抑制作用,其机制可能与抑制脂质过氧化等作用有关。更多还原

【Abstract】 Liver, which receive blood from portal vein, is the largest metabolic organ in the body. Faced with many insults such as ethanol, drugs, toxic agents, virus and bacteria, liver, which the number and state of activation of immunocyte resident in, is of much more difference compared with other immune organ. Severe inflammatory and injury can be initiated when faced with certain etiological factors, drug overdose, alcoholic intoxication, liver transplantation, fulminant hepatitis, extensive trauma, endotoxemia shock for instance,which leds to high mortality for disseminated intravascular coagulation and for multiple organ dysfunction syndrome during the acute inflammatory phase. The high mobidity of hepatitis is the main reason. Pathology showed hepatocyte degeneration, apoptosis and necrosis that are common causes of various liver diseases. Chronic liver injury can result in lose of its origin structure, gradually develop to cirrhosis and liver cancer. Therefore, positive and timely response to a variety of factors that cause liver damage will have important clinical significance.OBJECTIVE To investigate the effects and mechanisms of paeoniflorin (Pae) on mice with immunological liver injury (ILI) induced by Bacillus Calmette-Guérin (BCG) plus lipopolysaccharide (LPS)METHODS The model of ILI were induced by BCG plus LPS in mice. The activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) as well as the concentration of nitric oxide (NO) in serum were assayed by spectrophotometry. Malondiadehyde (MDA) content, superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities in liver homogenate were tested by the same method. ILI rats were induced by BCG plus LPS, too. Serum ALT and AST activities were assayed by the method mentioned above. The morphology changes in co-culture were observed on the 7th day. The effects of KC culture medium supplemented with different concentrations of Pae on HSC proliferation were detected by MTT assay. Hepatic tissue sections were stained with hematoxylin and eosin and examined under a light microscope.RESULTS (1) Pae (53,105,210 mg·kg-1) and TGP (85 mg·kg-1) ig were able to reduce mortality of ILI mice induced by BCG plus LPS. (2) Liver index, spleen index, and serum transaminase activities (ALT, AST) in ILI mice treated with various dose of Pae were significantly decreased compared with their counterpart in the model group. The area and extent of necrosis as well as the infiltration of inflammatory cells in hepatic tissue sections of ILI mice were attenuated. (3) KC supernatants could promote the HSC proliferation. KC supernatants treated with Pae could decrease the promoting effects of KC on HSC proliferation.CONCLUSION Paeoniflorin can ameliorate ILI in mice and decrease effects of KC on promoting HSC proliferation. The underlying mechanism may correlated with its ability of free radical scavenging, increasing the activities of SOD and GSH-Px.更多还原