抗OPN抗體干預(yù)沙鼠肝泡球蚴組織細(xì)胞因子長(zhǎng)期觀察結(jié)果及意義
本文關(guān)鍵詞: 骨橋蛋白 抗體 泡狀棘球蚴 侵襲性生長(zhǎng) 轉(zhuǎn)移 出處:《石河子大學(xué)》2013年碩士論文 論文類型:學(xué)位論文
【摘要】:目的:建立沙鼠肝泡狀棘球蚴動(dòng)物模型,外源性給予抗OPN抗體,根據(jù)在不同生長(zhǎng)階段,沙鼠肝泡球蚴及轉(zhuǎn)移組織中IL-2、IL-5、IL-10及TNF-α因子的表達(dá)水平,來(lái)探討這四種因子在沙鼠肝泡狀棘球蚴侵襲性生長(zhǎng)和轉(zhuǎn)移中的作用,觀察抗OPN抗體對(duì)泡狀棘球蚴生長(zhǎng)發(fā)育的影響。 方法:選擇4-6周齡健康長(zhǎng)爪沙鼠120只,雌性,體重(30±5)g。隨機(jī)分為三組,采用開(kāi)腹直視下肝臟穿刺接種多房棘球蚴原頭節(jié)混懸液的方法(0.15ml/只),建立肝泡狀棘球蚴動(dòng)物模型。一組單純接種泡狀棘球蚴組(模型組),二組在接種泡狀棘球蚴同時(shí)予以抗OPN抗體干預(yù)(實(shí)驗(yàn)組),三組在接種泡狀棘球蚴同時(shí)予以兔血清干預(yù)(對(duì)照組)。分別將三組接種泡狀棘球蚴的沙鼠隨機(jī)分成6小組,每小組10只,分別為感染140天、180天、220天、260天組。于相應(yīng)時(shí)間處死各組沙鼠,觀察泡狀棘球蚴的生長(zhǎng)、轉(zhuǎn)移等情況;免疫組化法測(cè)定肝泡狀棘球蚴及轉(zhuǎn)移組織標(biāo)本中的IL-2,IL-5,IL-10及TNF-α因子的表達(dá)。 結(jié)果: 1.140天、180天、220天組時(shí)實(shí)驗(yàn)組與對(duì)照組、實(shí)驗(yàn)組與模型組囊濕重有顯著性差異(F值分別為8.647,36.96,9.12,p值均小于0.05),而對(duì)照組與模型之間比較無(wú)差異(P㧐0.05)。 2.觀察泡狀棘球蚴轉(zhuǎn)移現(xiàn)象,發(fā)現(xiàn)肺轉(zhuǎn)移灶較少見(jiàn),而胸廓淋巴結(jié)轉(zhuǎn)移率較高,未見(jiàn)腦轉(zhuǎn)移灶。統(tǒng)計(jì)結(jié)果顯示:140天、180天、組時(shí)實(shí)驗(yàn)組與對(duì)照組、實(shí)驗(yàn)組與模型組胸廓淋巴結(jié)轉(zhuǎn)移率有顯著性差異(P㩳0.05),220天組時(shí)實(shí)驗(yàn)組與對(duì)照組,實(shí)驗(yàn)組與模型組胸廓淋巴結(jié)轉(zhuǎn)移率無(wú)顯著性差異(P㧐0.05)。 3. HE染色結(jié)果顯示:在140天、180天、220天時(shí)間段,實(shí)驗(yàn)組與對(duì)照組、實(shí)驗(yàn)組與模型組間E.m囊壁病理的改變無(wú)顯著性差異(P㧐0.05)。 4.免疫組化染色結(jié)果顯示:肝泡型棘球蚴組織中可見(jiàn)IL-2,IL-5, IL-10及TNF-α因子不同程度的表達(dá),定位于細(xì)胞漿,主要分布在肝泡狀棘球蚴纖維囊壁的生發(fā)層,細(xì)胞漿中可見(jiàn)黃色、棕黃色或棕褐色等陽(yáng)性顆粒;140天、180天組時(shí)IL-5,IL-10的表達(dá)在實(shí)驗(yàn)組與對(duì)照組、實(shí)驗(yàn)組與模型組比較有顯著性差異(P㩳0.05);140天、180天組時(shí)時(shí)間段,實(shí)驗(yàn)組與對(duì)照組、實(shí)驗(yàn)組與模型組IL-2及TNF-α的陽(yáng)性細(xì)胞率比較無(wú)顯著性差異(P㧐0.05),220天時(shí)時(shí)間段,,實(shí)驗(yàn)組與對(duì)照組、實(shí)驗(yàn)組與模型組IL-2,IL-5,IL-10及TNF-α的陽(yáng)性細(xì)胞率比較無(wú)顯著性差異。 結(jié)論:IL-2,IL-5,IL-10及TNF-α共同參與了宿主與肝泡狀棘球蚴相互間中晚期免疫進(jìn)程。外源性給予抗OPN抗體可使肝泡狀棘球蚴組織中IL-5,IL-10的表達(dá)減低,這可能是抑制肝泡狀棘球蚴中晚期侵襲性生長(zhǎng)和轉(zhuǎn)移的機(jī)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- 偽 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 鹵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- 偽 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- 偽 in hepatic alveolar hydatid tissues was observed by immunohistochemical staining. The expression of IL-2 IL-5, IL-10 and TNF- 偽 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- 偽 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- 偽 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- 偽 in the liver alveolar hydatid tissues can be decreased by exogenous anti OPN antibody, and the IL-10 and TNF- 偽 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.
【學(xué)位授予單位】:石河子大學(xué)
【學(xué)位級(jí)別】:碩士
【學(xué)位授予年份】:2013
【分類號(hào)】:R383.33
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