不同剂量泡球蚴感染所致小鼠肝脏病理变化及其免疫状态的研究

发布时间：2018-04-02 21:27

本文选题：泡球蚴　切入点：病理类型　出处：《新疆医科大学》2017年硕士论文

【摘要】：目的:泡型包虫病(Alveolar Echinococcosis,AE)是由泡球蚴(Ec-hinococcus multilocularis,Em)寄生人体,导致慢性浸润性的肝脏损伤为特点的寄生虫病。(1)通过观察不同感染时间剂量下泡球蚴对小鼠肝脏所造成的病理损伤、病灶数量和类型的变化;(2)评估在不同感染剂量下,泡球蚴对小鼠肝脏造成的损伤程度;(3)检测不同剂量泡球蚴感染条件下CD4+T细胞亚群的变化及炎症因子基因表达,从而分析宿主不同免疫状态下对泡球蚴寄生的影响。方法:(1)给小鼠5种剂量(50psc,250psc,500psc,1000psc,2000psc)泡球蚴达到感染泡球蚴的目的,分别在7个时间段(2周、4周、8周、12周、16周、20周、24周)采集小鼠肝脏标本,制备H E染色片,计算不同时间和剂量下小鼠肝脏病灶数目和类型;(2)选取3种剂量(50psc,500psc,2000psc),3个感染时间段(2周、12周、24周)小鼠的肝脏的切片进行α-SMA,RS,ki67染色,评估肝脏纤维化水平以及淋巴细胞增殖水平;(3)分离肝脏淋巴细胞,检测肝脏区域CD4+T细胞亚群的变化和相关炎性因子基因的表达水平。结果:(1)高剂量(2000psc,1000psc)的泡球蚴产生的病灶数量显著高于低剂量组(P0.05),病灶数量不会随着感染时间延长而减少,反而会增多。并且,病灶面积大,泡球蚴对肝脏侵犯严重。但是,低剂量(50psc,250psc)以及中等剂量(500psc)的泡球蚴感染小鼠后,随着感染时间延长,病灶可以被宿主清除,病灶所引起的局部肝损伤也可以恢复;(2)高剂量组α-SMA以及RS染色的阳性率显著高于低剂量组(P0.05);2000psc剂量感染后,分别在2周、24周COL1A1和CO L3A1的表达显著高于正常对照组(P0.05);(3)250psc在不同感染时间点Th1/Th2并无明显变化,而500psc剂量Th1/Th2在感染24周时显著升高(P0.05)。2000p sc在24周时Th1/Th2有显著下降(P0.05);(4).高剂量组和低剂量组在感染晚期(24周)都表现为Treg/Th17平衡向Th17偏移。结论:(1)低剂量泡球蚴感染小鼠,其病灶可以被宿主清除;但是,高剂量泡球蚴感染小鼠,其病灶不能被清除;(2)高剂量泡球蚴感染小鼠可造成更严重的肝纤维化;(3)宿主免疫类型偏向Th1与Th17可能有利于病灶清除。
[Abstract]:Objective: alveolar echinococcosis (Alveolar Echinococcosis, AE) by Echinococcus multilocularis (Ec-hinococcus multilocularis, Em) parasitic body, leading to liver infiltrating the chronic injury to parasitic disease characteristics. (1) the pathological injury by observing the different infection time dose of Echinococcus multilocularis in mice liver caused by changes in the number of dirty. And the type of lesions; (2) in the evaluation of different levels of infection, the extent of damage caused by Echinococcus multilocularis in liver of mice; (3) the change of inflammatory cytokines and gene expression of CD4+T cell subsets and testing different doses of Echinococcus multilocularis infection conditions, so as to analyze the different immune status of the host of Echinococcus multilocularis parasitic effects methods: (1) to the mice of 5 doses (50psc, 250psc, 500psc, 1000psc, 2000psc) of Echinococcus multilocularis infection of Echinococcus multilocularis purpose, in 7 time periods (2 weeks, 4 weeks, 8 weeks, 12 weeks, 16 weeks, 20 weeks, 24 weeks) acquisition of mouse liver preparation of H E staining specimens. Time and dose, different number of mouse liver lesions and the type of calculation; (2) selected 3 doses (50psc, 500psc, 2000psc), with 3 time period (2 weeks, 12 weeks, 24 weeks) in mice liver slices of alpha -SMA, RS, Ki67 staining, assessment of liver fibrosis level lymphocyte proliferation and level; (3) to detect the expression of lymphocytes isolated from the liver, liver area changes of CD4+T cell subsets and related inflammatory factor gene. Results: (1) high dose (2000psc, 1000psc) of Echinococcus multilocularis generated by the number of lesions was significantly higher than that in the low dose group (P0.05), number of lesions and not decreased with the prolongation of infection, but will increase. And the lesion area, the liver Alveococcus seriousviolation. However, low dose (50psc, 250psc), medium dose (500psc) of Echinococcus multilocularis infection in mice after infection with the time prolonged, lesions can be eliminated by host, lesions caused by Partial liver injury can be restored; (2) the positive rate of the high dose group of alpha -SMA and RS staining was significantly higher than the low dose group (P0.05); the dose of 2000psc after infection, respectively, in 2 weeks, 24 weeks and CO COL1A1 expression of L3A1 was significantly higher than that of normal control group (P0.05); (3) 250psc in the same at the time of Th1/Th2 infection did not change significantly, while the 500psc dose of Th1/Th2 infection was significantly increased at 24 weeks (P0.05).2000p SC at 24 weeks Th1/Th2 was significantly decreased (P0.05); (4). High dose group and low dose group in the later stage of infection (24 weeks) showed Treg/Th17 equilibrium shift to Th17. Conclusion: (1) low dose of Echinococcus multilocularis infected mice, the lesions can be eliminated by host; however, high doses of Echinococcus multilocularis infected mice, the lesions cannot be removed; (2) the high dose of Echinococcus multilocularis infected mice can cause more severe hepatic fibrosis; (3) the host immune shows type Th1 and Th17 may is conducive to debridement.

【学位授予单位】：新疆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R532.32

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