枯否细胞在非酒精性脂肪肝炎发生中的作用-病理生理学专业毕业论文.docxVIP

山西医科大学摘 山西医科大学 摘 要 枯否细胞在非酒精性脂肪肝炎 发生中的作用 导师许瑞龄教授 研究生吴惠文 目的：探讨枯否细胞(Kupffer cell，KCS)在高脂饮食致 小鼠非酒精性脂肪肝炎发生中的作用。 方法：选取雌性昆明种小鼠46只，随机分为两组，高 脂组与对照组。高脂组给予高脂饮食(基础饲料+1 5％玉米 油+5％胆固醇)，对照组饲以正常饮食。l 2周后每组取小 鼠10只，1％戊巴比妥钠麻醉，腹主动脉采血，用于血浆 ALT、LPS的测定。取肝、脾并称重，计算肝、脾系数。肝 右叶罱于1 0％的中性福尔马林固定用于HE染色切片。其 余肝组织用于MDA、TG、TNF．Q水平检测。另取小鼠7 只，在尾静脉注射印度墨汁后眼眶采血计算KCs的吞噬指 数。剩余小鼠用做KCs的分离培养，以探讨NASH时KCs 吞噬聚苯乙烯乳珠的能力及其在内毒素刺激下分泌TNF—o 的变化。与此同时采用WeStern b10t方法研究肝组织CD68 及KCS CDl4的表达。 结果：小鼠在高脂饮食1 2周后，肝系数增加。组织病 理学显示小鼠肝细胞内有大量的脂肪空泡，Malory小体的 形成及大量的炎性细胞浸润，表明小鼠已发生非酒精性脂肪 肝炎。血浆ALT、LPS、肝匀浆MDA、TG、及TNF．a水平 均明显高与对照组(PO．05)。而尾静脉注射印度墨汁后其清 除碳粒能力则低于对照组(P0．05)。 l(Cs分离培养的实验结果表明，NASH组KCs呈活化状 山西医科走学态，但其吞噬聚苯乙烯乳珠能力降低，而给予LPS 山西医科走学 态，但其吞噬聚苯乙烯乳珠能力降低，而给予LPS (1 00ng／m1)刺激4h后TNF—d的分泌则增加(PO．05)。 WeSt en blOt结果表明，NAsH组肝组织CD68及KCS的 CDl 4表达上调。 结论：在高脂饮食所致小鼠发生NASH时，虽KCS可被 激活，但其吞噬功能减低、而分泌TNF—a增加。提示KCs 功能紊乱与NASH发生密切相关。 关键词KCs；NAStl；TNF～o；吞噬功能；甘油三酯 LPS -2- 山西医科走举 山西医科走举 硕士学位论文2003 ABSTRACT The role of kupffer cells in the pathogenesis of nonalcoholic steatohepati“s OBJECTION To explore the role of kupffer cells(KCs)in the pathogenesis of mice given high—fat diet to nonalcoholic steatohepatitis． METHODE In this experiments，46 female mice were randomly divided into two groups．Model group mice were given high—fat diet(basic diet +1 5％corn oil+5％cholesterol、and the control group were given normal diet．After 1 2 weeks，ten mice each group were sacrificed under 1％ pentobarbital sodium anesthesia，blood sample were collected from abdominal vein for measuring ALT and LPS．Liver sections and spleen were weighted to calculated liver coefficient and spleen coefficient Left liver was kept in 1 0％formalin for making paraffin sections and the rest was stored at-80。C until assays for TG，TNF—d，MDA．7 mice each group were injected Indian ink from tail vein for calculating phagocytose index． The rest mice were used as KCs isolated to assess function of KC’s phagocytose and ability of KCs’secretion after LPS stimulation．Westen blottoanalyzeliverCD68and KCs’CDl4