本文選題：大片形吸蟲 + 囊蚴培養(yǎng)�。� 參考：《廣西大學(xué)》2015年碩士論文

【摘要】：片形吸蟲病主要侵害宿主的肝膽系統(tǒng),且該病早期診斷困難,即便后期用藥驅(qū)除了蟲體,對肝臟造成的損傷也較難修復(fù)。因此探明大片形吸蟲感染后肝臟損傷的相關(guān)機(jī)制,對豐富大片形吸蟲和宿主之間的相互作用機(jī)制及感染宿主的肝臟治療和康復(fù)具有極其重要的意義。本研究內(nèi)容包括：大片形吸蟲囊蚴實(shí)驗(yàn)室繁育方案的優(yōu)化完善,大片形吸蟲-昆明小鼠感染模型的建立以及感染昆明小鼠肝臟病理學(xué)的初步研究；這些研究旨在為大片形吸蟲肝臟免疫和組織損傷修復(fù)等下游研究夯實(shí)基礎(chǔ)。首先完善優(yōu)化實(shí)驗(yàn)室人工培養(yǎng)大片形吸蟲囊蚴體系,并對大片形吸蟲在中間宿主體內(nèi)的發(fā)育特點(diǎn)進(jìn)行觀察。分別以1,3,7條毛蚴感染2～3mm,5～7mm兩種體型大小的小土蝸螺,在室溫26℃時(shí),感染后10～17天可見母雷蚴,28天可見子雷蚴體內(nèi)充滿尾蚴,30～33天可見尾蚴釋放并結(jié)囊為囊蚴。對比分析不同大小的螺感染不同數(shù)量的毛蚴最后產(chǎn)囊蚴率結(jié)果,得出最佳培養(yǎng)方案與肝片形吸蟲囊蚴培養(yǎng)不同,大片形吸蟲囊蚴培養(yǎng)選用3-5mm大小的螺,每螺感染3條毛蚴時(shí)產(chǎn)囊蚴量最多。以5、15、30和50個(gè)囊蚴經(jīng)口感染小鼠,于不同時(shí)期分別剖檢以觀察蟲體分布、回收、發(fā)育情況,以及肝臟的病理變化。結(jié)果顯示：感染后5天內(nèi)便可在肝實(shí)質(zhì)探查到蟲體,35-42天內(nèi)童蟲完成從肝實(shí)質(zhì)向膽管移行,但各感染組均未發(fā)現(xiàn)成蟲；大片形吸蟲在昆明小鼠體內(nèi)異位寄生現(xiàn)象發(fā)生頻率較高,肺部、口腔、膈肌、前肢皮下等部位均可發(fā)現(xiàn)蟲體；受感染昆明小鼠肝臟可見彌散出血、化膿、脂變、纖維化、壞死、硬變等明顯肉眼病理變化,病變程度隨感染強(qiáng)度的增加而加重。表明大片形吸蟲-昆明小鼠模型適宜用于童蟲期的研究,15個(gè)囊蚴的感染強(qiáng)度是動態(tài)觀察昆明小鼠宿主狀態(tài)及相關(guān)研究的適宜感染劑量；小鼠為大片形吸蟲的非適宜宿主,不宜作為研究反芻動物(適宜宿主)感染大片形吸蟲動物模型。觀察人工感染大片形吸蟲小鼠的肝臟組織學(xué)病理變化顯示：感染后7天,肝臟肉眼可見輕微損傷,HE染色可見肝細(xì)胞排列開始紊亂；感染后21～42天,肝臟損失逐漸加重,顯微鏡下可觀察到肝細(xì)胞壞死、結(jié)締組織增生、炎性細(xì)胞的增生浸潤等典型的肝組織病理學(xué)改變。檢測感染大片形吸蟲的小鼠血清中堿性磷酸酶(ALP)、谷草轉(zhuǎn)氨酶(AST)、谷丙轉(zhuǎn)氨酶(ALT)、總蛋白(TP)、白蛋白(ALB)、球蛋白(GLOB)及白球比(A/G)的變化,顯示AST在感染后第3天開始升高,21～56天顯著升高(P0.01),49天達(dá)峰值；ALT于感染后14天開始升高,21～35天開始顯著升高(P0.01),49天達(dá)峰值；ALP呈現(xiàn)感染后14～21天升高,35～49天下降,56天又開始回升的波動變化;GLOB于感染后21～56天總體呈上升趨勢；ALB自感染后呈現(xiàn)下降趨勢,感染后49天達(dá)最低值；表明：血清肝臟損失相關(guān)因子的變化與肝臟組織病變動態(tài)發(fā)展相一致；AST.ALT可作為感染急性期指標(biāo),ALB.GLOB和A/G比值的范圍可評估大片形吸蟲病所引起的肝損傷程度,聯(lián)合檢測這些生化指標(biāo)對大片形吸蟲病診斷及預(yù)后具有重要意義。
[Abstract]:Fasciosis mainly infringes the liver and bile system of the host, and it is difficult to diagnose the early stage of the disease. Even if the later drug expelling the insect body, the damage to the liver is difficult to repair. Treatment and rehabilitation are of great significance. This study includes the optimization of laboratory breeding programs for large Fasciola cercariae, the establishment of large Fasciola - Kunming mice infection model and the preliminary study of liver pathology in Kunming mice; these studies are aimed at repairing the liver immunity and tissue damage of large Fasciola. In the first step, we should improve the foundation of the study of the lower reaches of the lower reaches. First, improve the artificial cultivation of large tracer cercariae in the laboratory, and observe the development characteristics of large tracts in the middle host. The 1,3,7 cercariae infect 2 to 3mm, 5 to 7mm, and two kinds of small snail size. At room temperature 26, the larvae can be seen for 10~17 days after infection. On the 28 day, the cercariae were full of cercariae in the cercariae, the cycariae were released on the 30~33 day and cysts were cysts. Compared and analyzed the results of the final production of cercariae in different numbers of different sizes of the cercariae, the best training scheme was different from the culture of the liver slice form of the cercariae. The 3-5mm size snails were selected for the cultivation of the large fasciolus cercariae, and 3 hairs per snails were infected. The amount of cercariae was the most. The mice infected with 5,15,30 and 50 cercariae were infected in different periods to observe the distribution, recovery, development, and pathological changes of the liver. The results showed that the insect body could be detected in the liver parenchyma within 5 days after infection, and in 35-42 days, the children migrated from the liver parenchyma to the bile duct, but all the infected groups were all infected. No adult worms were found. The occurrence of ectopic parasitism in Kunming mice was high, and the body was found in the lungs, the mouth, the diaphragm and the subcutaneous parts of the forelimb. The liver of the infected Kunming mice showed diffuse hemorrhage, pyogenic, fat, fibrosis, necrosis and hard change, and the degree of pathological changes increased with the intensity of infection. It was added and aggravated. It showed that the model of large bugs - Kunming mice was suitable for the study of the stage of childhood. The infection intensity of the 15 cycariae was a dynamic observation of the host state of Kunming mice and the appropriate dose of infection. Histopathological changes of liver in mice with artificial infection of large tracts showed that 7 days after infection, the liver was slightly damaged in the naked eye, the liver cell arrangement began to be disorganized by HE staining, and the liver damage was gradually increased on the 21~42 day after infection. The necrosis of hepatocytes, connective tissue proliferation and inflammatory cells increased under the microscope. The changes in serum alkaline phosphatase (ALP), glutamic pyruvic aminotransferase (AST), glutamic pyruvic aminotransferase (ALT), total protein (TP), albumin (ALB), globulin (GLOB) and white ball ratio (A/G) in mice infected with Fasciola Fasciola showed that AST increased at third days after infection and increased significantly (P0.01), 49 (P0.01), 49. The peak value of Tianda, ALT began to rise on the 14 day after infection, the 21~35 days began to rise significantly (P0.01), 49 Tianda peak, ALP showed an increase of 14~21 days after infection, 35~49 days, and the 56 days began to rebound, and the overall increase trend was 21~56 days after infection; ALB showed a downward trend after infection and the lowest value of 49 Tianda after infection. The changes in serum liver loss related factors are consistent with the dynamic development of liver tissue lesions, and AST.ALT can be used as an indicator of acute infection, and the range of ALB.GLOB and A/G ratios can be used to assess the degree of liver damage caused by large Fasciola. The combined detection of these biochemical indexes is important for the diagnosis and prognosis of large Fasciola. Righteousness.
【學(xué)位授予單位】：廣西大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S855.99

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本文編號：2117815