小鼠日本血吸虫感染中AIF-1的动态表达及其干预的影响

发布时间：2018-04-11 06:44

本文选题：AIF-1 + 日本血吸虫　；参考：《华中科技大学》2011年博士论文

【摘要】：AIF-1，即同种异体移植物炎症因子-1（allograft inflammatory factor-1），是Utans最先于1995年在大鼠和人异体移植心脏组织中发现的一种蛋白质。随后的研究发现AIF-1存在于从红海鲷、鲤鱼和海绵等低等动物低到鼠、猪、牛、人等高等哺乳动物中，且种属间高度保守。AIF-1是一种早期炎症因子，在多种与炎症反应相关的病变中表达升高，具有非常复杂、广泛而重要的生物学功能。 AIF-1在多种人体自身免疫性疾病、器官移植和实验性免疫性疾病等组织中过表达。有人发现AIF-1的过表达可以促进免疫反应由Th1型向Th2型倾斜；另有人发现AIF-1转基因小鼠的肠炎病变较未转基因小鼠的病变轻，并认为这是由于高表达的AIF-1反馈性抑制Th1型的免疫反应所致。现有研究表明，AIF-1通过与巨噬细胞、T淋巴细胞、内皮细胞和TNF-α, IFN-γ, TGF-β等相互影响、相互作用，并通过影响机体的免疫调节系统而发挥其重要功能。血吸虫感染是一个很复杂的免疫反应性炎症过程，由其导致的肝纤维化、肝硬化严重威胁人类健康和经济发展。我们推测，与其他免疫炎症性病变一样，感染血吸虫的宿主的肝、脾等组织中AIF-1表达增强，而且如果用AIF-1纯化物干预感染小鼠，还可对疾病的进程产生明显影响。因此，我们设计、实施本课题，并初步证实了我们的设想。本实验分二部分：第一部分：首次证实感染血吸虫的BALB/c小鼠肝组织有不同程度的AIF-1过表达，以急性期最强，随后逐渐下降。ELISA检测受SEA刺激72小时的脾淋巴细胞培养上清液中AIF-1的浓度，发现各感染组均明显低于正常对照小鼠；同时检测TNF-α和TGF-β的表达，发现这3种细胞因子的表达变化有一定的量变关系。这些结果提示我们，AIF-1是一种早期炎症因子，与TNF-α和TGF-β等细胞因子相互影响，在血吸虫肝病发生发展进程中发挥重要作用。第二部分：AIF-1纯化物干预血吸虫感染小鼠，并设立对照组，发现AIF-1干预组在感染后5周时肝细胞损害稍加重，感染后8周肝组织坏死和纤维化均较对照组减轻，以14周时差异最明显；Western blot，RT-PCR和ELISA检测发现干预组AIF-1表达随感染进展而逐渐升高；这表明AIF-1干预导致血吸虫慢性感染期小鼠体内有效的AIF-1浓度增加，且对肝脏具有保护作用，说明AIF-1纯化物能有效预防和治疗慢性血吸虫肝病。如何选择更有效的干预时间和剂量，是我们下一步要研究的课题。除此以外，由于各种原因导致的纤维化发生的细胞、分子机制相似，深入研究AIF-1抗血吸虫病肝纤维化作用的分子机理，可为有效防治其他类似的纤维化病变（如系统性硬化症、类风湿性关节炎等）提供理论依据。综上所述，本研究首次证实： 1.日本血吸虫感染所致BALB/c小鼠肝病组织过表达AIF-1，以感染急性期最强，后逐渐下降。 2. AIF-1是一种早期炎症因子，与TNF-α和TGF-β等细胞因子紧密联系并相互作用，在血吸虫肝病的发生发展中发挥重要的作用。 3. AIF-1纯化物干预导致血吸虫慢性感染期小鼠体内有效AIF-1浓度增加，而其肝组织坏死和纤维化反而明显减轻。 4. AIF-1纯化物干预能有效预防和治疗慢性血吸虫肝病。
[Abstract]:It was found in the rat , pig , cattle , human and other high mammals , and highly conserved among the species in the low - grade mammals , such as red sea bream , carp and sponge , and increased in various diseases related to inflammatory response , which had a very complex , broad and important biological function .

It has been found that the overexpression of protein - 1 can promote the immune response from Th1 type to Th2 type .
In addition , it was found that the disease of transgenic mice was less than that of non - transgenic mice , and it was considered to be a result of the high expression of the anti - feed inhibition of Th1 type immune response . The existing study showed that the interaction of the protein with macrophages , T lymphocytes , endothelial cells and TNF - 伪 , IFN - 纬 , TGF - 尾 , and the like , and played an important function by affecting the immune regulation system of the organism .

Schistosoma japonicum infection is a very complicated process of immune response inflammation caused by hepatic fibrosis and cirrhosis , which is a serious threat to human health and economic development .

In the first part , the BALB / c mouse liver tissues infected with Schistosoma japonicum were first confirmed to be overexpressed in different degrees , the strongest in the acute phase and then gradually decreased .
At the same time , the expression of TNF - 伪 and TGF - 尾 was detected , and it was found that the expression changes of these three cytokines have a certain amount . These results suggest that the expression of TNF - 伪 and TGF - 尾 is a kind of early inflammatory factor , which interacts with cytokines such as TNF - 伪 and TGF - 尾 , and plays an important role in the development of liver disease of Schistosoma japonicum .

In the second part , the mice infected with Schistosoma japonicum were infected with the purified protein , and the control group was established . It was found that the hepatocyte injury was slightly increased at 5 weeks after infection , and the necrosis and fibrosis of the liver tissues in 8 weeks after infection were less than those of the control group , and the difference was the most obvious at 14 weeks .
Western blot , RT - PCR and ELISA were used to detect the expression of protein in the intervention group and gradually increase with the progression of infection .
These results suggest that the effective anti - inflammatory activity in the mice infected with Schistosoma japonicum is increased and the protective effect on the liver is indicated , and it is indicated that the purified product can effectively prevent and treat the chronic schistosomiasis liver disease .

How to select more effective intervention time and dosage is the subject of our next step to study . In addition , the molecular mechanism similar to the cell and molecular mechanism caused by various causes is similar , so the molecular mechanism of anti - schistosomiasis liver fibrosis is deeply researched , which can provide theoretical basis for the effective prevention and treatment of other similar fibrotic diseases ( such as systemic sclerosis , rheumatoid arthritis , etc . ) .

In summary , the present study confirms the first time :

1 . The liver disease of BALB / c mice caused by Schistosoma japonicum infection has been overexpressed in BALB / c mice , which is the strongest in the acute phase of infection , and then gradually decreases .

2 . It is an early inflammatory factor that closely links and interacts with cytokines such as TNF - 伪 and TGF - 尾 and plays an important role in the development of liver disease of Schistosoma japonicum .

3 . There was a significant increase in the effective amount of protein - 1 in the body of mice with chronic infection of Schistosoma japonicum , but the necrosis and fibrosis of liver tissue were obviously reduced .

4 . Treatment of chronic schistosomiasis liver disease can be effectively prevented and treated by the intervention of purified protein .

【学位授予单位】：华中科技大学
【学位级别】：博士
【学位授予年份】：2011
【分类号】：R383.24

【参考文献】