抗OPN抗体干预沙鼠肝泡球蚴组织细胞因子长期观察结果及意义

发布时间：2018-02-28 00:42

本文关键词： 骨桥蛋白抗体泡状棘球蚴侵袭性生长转移　出处：《石河子大学》2013年硕士论文　论文类型：学位论文

【摘要】：目的：建立沙鼠肝泡状棘球蚴动物模型，外源性给予抗OPN抗体，根据在不同生长阶段，沙鼠肝泡球蚴及转移组织中IL-2、IL-5、IL-10及TNF-α因子的表达水平，来探讨这四种因子在沙鼠肝泡状棘球蚴侵袭性生长和转移中的作用，观察抗OPN抗体对泡状棘球蚴生长发育的影响。方法：选择4-6周龄健康长爪沙鼠120只，雌性，体重（30±5）g。随机分为三组，采用开腹直视下肝脏穿刺接种多房棘球蚴原头节混悬液的方法(0.15ml/只)，建立肝泡状棘球蚴动物模型。一组单纯接种泡状棘球蚴组（模型组），二组在接种泡状棘球蚴同时予以抗OPN抗体干预（实验组），三组在接种泡状棘球蚴同时予以兔血清干预（对照组）。分别将三组接种泡状棘球蚴的沙鼠随机分成6小组，每小组10只，分别为感染140天、180天、220天、260天组。于相应时间处死各组沙鼠，观察泡状棘球蚴的生长、转移等情况；免疫组化法测定肝泡状棘球蚴及转移组织标本中的IL-2，IL-5，IL-10及TNF-α因子的表达。结果： 1.140天、180天、220天组时实验组与对照组、实验组与模型组囊湿重有显著性差异（F值分别为8.647，36.96，9.12，p值均小于0.05），而对照组与模型之间比较无差异（P㧐0.05）。 2.观察泡状棘球蚴转移现象，发现肺转移灶较少见，而胸廓淋巴结转移率较高，未见脑转移灶。统计结果显示：140天、180天、组时实验组与对照组、实验组与模型组胸廓淋巴结转移率有显著性差异（P㩳0.05），220天组时实验组与对照组，实验组与模型组胸廓淋巴结转移率无显著性差异（P㧐0.05）。 3. HE染色结果显示:在140天、180天、220天时间段，实验组与对照组、实验组与模型组间E.m囊壁病理的改变无显著性差异（P㧐0.05）。 4.免疫组化染色结果显示：肝泡型棘球蚴组织中可见IL-2，IL-5， IL-10及TNF-α因子不同程度的表达，定位于细胞浆，主要分布在肝泡状棘球蚴纤维囊壁的生发层，细胞浆中可见黄色、棕黄色或棕褐色等阳性颗粒；140天、180天组时IL-5，IL-10的表达在实验组与对照组、实验组与模型组比较有显著性差异（P㩳0.05）；140天、180天组时时间段，实验组与对照组、实验组与模型组IL-2及TNF-α的阳性细胞率比较无显著性差异（P㧐0.05），220天时时间段，，实验组与对照组、实验组与模型组IL-2，IL-5，IL-10及TNF-α的阳性细胞率比较无显著性差异。结论：IL-2，IL-5，IL-10及TNF-α共同参与了宿主与肝泡状棘球蚴相互间中晚期免疫进程。外源性给予抗OPN抗体可使肝泡状棘球蚴组织中IL-5，IL-10的表达减低，这可能是抑制肝泡状棘球蚴中晚期侵袭性生长和转移的机制之一。
[Abstract]:Objective: to establish an animal model of hepatic alveolar echinococcosis in gerbils, and give exogenous anti OPN antibody. According to the expression levels of IL-2, IL-5, IL-10 and TNF- 伪 in liver alveolar hydatid and metastatic tissues of gerbils at different growth stages. To investigate the role of these four factors in the invasive growth and metastasis of hydatid alveolar hydatid in gerbils, and to observe the effect of anti OPN antibody on the growth and development of hydatid alveolar hydatid. Methods: 120 healthy gerbils aged 4-6 weeks, female, weight 30 卤5 g, were randomly divided into three groups. The animal model of hepatic alveolar echinococcosis was established by inoculating the mixed suspension of multilocular echinococci with liver puncture under open and open vision. One group was inoculated with alveolar echinococcus group (model group) and the other group was inoculated with alveolar echinococci (Echinococcus alveolata) group (model group and two groups were inoculated with alveolar echinococcus). The three groups were inoculated with alveolar echinococcus and treated with rabbit serum at the same time (control group). The three groups of gerbils inoculated with alveolar echinococcus were randomly divided into 6 groups. Ten rats in each group were divided into two groups: group A (140 days) and day 180 (180 days) and day 220 (day 260). The gerbils in each group were killed at the corresponding time to observe the growth and metastasis of alveolar echinococcus. The expression of IL-2IL-5, IL-10 and TNF- 伪 in liver alveolar echinococci and metastatic tissues were detected by immunohistochemical method. Results:. 1.140 days and 180 days after 220 days, there was significant difference between the experimental group and the control group. The F values of the experimental group and the model group were 8.647 ~ 36.96 ~ 9.12p < 0.05, respectively, but there was no significant difference between the control group and the model group (P > 0.05). 0.05. 2. The phenomenon of alveolar hydatid metastasis was observed. The pulmonary metastasis was rare, but the thoracic lymph node metastasis rate was higher than that of the control group. There was significant difference in the rate of thoracic lymph node metastasis between the experimental group and the model group. There was no significant difference in the rate of thoracic lymph node metastasis between the experimental group and the control group. 0.05. 3. The results of HE staining showed that there was no significant difference between the experimental group and the control group, the experimental group and the model group in the pathological changes of the E m cyst wall in the period of 140 days, 180 days and 220 days, and there was no significant difference in the pathological changes between the experimental group and the control group. 0.05. 4. The expression of IL-2 IL-5, IL-10 and TNF- 伪 in hepatic alveolar hydatid tissues was observed by immunohistochemical staining. The expression of IL-2 IL-5, IL-10 and TNF- 伪 was localized in the cytoplasm, mainly distributed in the germinal layer of the fibrous cyst wall of hepatic alveolar echinococcus, and yellow was observed in the cytoplasm. The expression of IL-5 and IL-10 in the experimental group and the control group were significantly different between the experimental group and the model group. The positive cell rates of IL-2 and TNF- 伪 in the experimental group and the control group were not significantly different from those in the control group and the model group. There was no significant difference in the positive rates of IL-5 and IL-10 and TNF- 伪 between the experimental group and the control group, the experimental group and the model group. Conclusion the expression of IL-5 IL-10 and TNF- 伪 in the liver alveolar hydatid tissues can be decreased by exogenous anti OPN antibody, and the IL-10 and TNF- 伪 are involved in the immune process of the host and liver alveolar echinococci, and the expression of IL-5 IL-10 in the liver alveolar hydatid tissues can be decreased by exogenous anti OPN antibody. This may be one of the mechanisms that inhibit the invasion and metastasis of hepatic alveolar echinococcus.
【学位授予单位】：石河子大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R383.33

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