中文题名: | 探究诱导大鼠炎性肝癌模型和免疫过程中尿蛋白组的动态变化 |
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保密级别: | 公开 |
论文语种: | 中文 |
学科代码: | 071000 |
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学生类型: | 硕士 |
学位: | 理学硕士 |
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学位年度: | 2022 |
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研究方向: | 探究诱导大鼠炎性肝癌模型和免疫过程中尿蛋白组的动态变化 |
第一导师姓名: | |
第一导师单位: | |
提交日期: | 2022-06-18 |
答辩日期: | 2022-06-18 |
外文题名: | To explore the dynamic changes of urinary proteome during Inflammatory liver cancer model induction and immunity in rats |
中文关键词: | |
外文关键词: | Proteomics ; Urine ; Inflammatory cancer model ; liver cancer ; Immune |
中文摘要: |
尿液更利于积累和反映机体生理状态的变化且不受到稳态机制的调节,是疾病早期生物标志物的良好来源。
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肝癌是非常常见的一种恶性肿瘤,其发病过程隐匿,病程时间短,发病率和死亡率却相对较高。通过DEN诱导的肝癌-炎癌模型,观察疾病不同阶段尿液蛋白质组动态变化过程。在第二章,我们腹腔注射DEN构建大鼠肝癌-炎癌模型,每周收集尿液,通过液相色谱联用质谱(LC-MS/MS)鉴定差异蛋白,使用IPA软件对差异蛋白进行生物学通路的分析,观察大鼠肝癌-炎癌模型中疾病不同阶段尿蛋白变化情况。15只实验大鼠通过DEN诱导,构建了从肝炎-肝硬化-肝癌疾病模型的不同时期,并且每只大鼠由于个体差异出现了疾病进展快慢不同的现象。每只大鼠在不同疾病时间点都出现了较多的差异蛋白,不同大鼠的差异蛋白都富集到了相同的与肝脏损伤、炎症病变有关、与肿瘤的发生有关、与肿瘤潜在的治疗靶点有关的生物学通路。 为更好地观察尿液中免疫系统不同的反应和变化。通过对大鼠注射牛血清白蛋白和氢氧化铝佐剂,观察尿液蛋白质组中免疫系统的变化。在第三章实验中对大鼠大腿肌肉注射牛血清白蛋白和氢氧化铝佐剂,收集尿液,通过液相色谱联用质谱(LC-MS/MS)鉴定差异蛋白,使用IPA软件对差异蛋白进行生物学通路的分析,观察大鼠尿蛋白中免疫系统的变化情况。15只大鼠分别肌肉注射生理盐水、氢氧化铝佐剂、牛血清白蛋白(BSA)、氢氧化铝佐剂和牛血清白蛋白混合物,构建对照组、佐剂组、BSA组、混合组模型。不同组别之间两两比较差异蛋白得到相关生物学通路,发现尿液中可以观察到佐剂帮助牛血清白蛋白更早的激发免疫系统发生反应。并且在尿液中还观察到混合组先后激发炎症反应、T细胞活化、抗原提呈细胞相关、B细胞相关等免疫相关通路。 以上研究分别展示了我们可以在肝癌-炎癌模型各阶段观察到尿液蛋白质组不同的变化,这些通路都被提及与肝炎、癌前病变、肝癌相关,甚至与癌症的治疗相关,这为寻找肝癌的生物标志物提供了线索,并且不同大鼠由于个体差异会出现疾病进展不同的现象,这提示我们之后需要更加注意检测的个体化、精准化;我们可以在早期从尿液蛋白质组中观察到免疫系统的变化,富集到一些与免疫应答过程中相关的生物学通路,并且可以区分开不同免疫强弱的实验组,这为以后免疫系统的研究提供了新思路,新方向,甚至加快疫苗的研发提供一些新的线索和依据。 |
外文摘要: |
Urine is more conducive to the accumulation and reflection of the body’s physiological state changes and is not regulated by the homeostasis mechanism, so it is a good source of biomarkers in the early stage of the disease. Liver cancer is a very common malignant tumor. Its pathogenesis is hidden, with a short duration and incidence rate and mortality rate relatively high. Through the liver cancer-inflammatory cancer model induced by DEN, observe the dynamic change process of urine proteome at different stages of the disease. In chapter two, a rat liver cancer-inflammatory cancer model was constructed by intraperitoneal injection of DEN, urine was collected weekly, and differential proteins were identified by liquid chromatography coupled with mass spectrometry (LC-MS/MS), and the biological pathways of differential proteins were analyzed using IPA software. To observe the changes in urine protein at different stages of the disease in the rat liver cancer-inflammatory cancer model. Fifteen experimental rats were induced by DEN to construct different stages of the hepatitis-cirrhosis-liver cancer disease model, and each rat had a different disease progression due to individual differences. Each rat has more differential proteins at different disease time points. The differential proteins of different rats are enriched in the same biological pathways related to liver injury, inflammatory lesions, tumor occurrence, and potential therapeutic targets of a tumor. To better observe the different reactions and changes of the immune system in the urine. The changes in the immune system in the urine proteome were observed by injecting bovine serum albumin and aluminum hydroxide adjuvant into rats. In chapter three, bovine serum albumin and aluminum hydroxide adjuvant were injected into rat thigh muscle, urine was collected, differential proteins were identified by liquid chromatography-mass spectrometry (LC-MS / MS), and biological pathways of differential proteins were analyzed by IPA software to observe the changes of the immune system in rat urinary protein. Fifteen rats were intramuscularly injected with normal saline, aluminum hydroxide adjuvant, bovine serum albumin, aluminum hydroxide adjuvant, and bovine serum albumin (BSA) mixture to construct the models of the control group, adjuvant group, BSA group, and mixed group. Comparing the different proteins between different groups to get the relevant biological pathways, it was found that adjuvants can be observed in urine to help bovine serum albumin stimulate the immune system to respond earlier. It was also observed in urine that the mixed group successively stimulated immune-related pathways such as inflammatory response, T cell activation, antigen-presenting cell-related pathways, and B cell-related pathways. The above studies show that we can observe different changes in urinary proteome in each stage of the liver cancer inflammatory cancer model. These pathways are mentioned to be related to hepatitis, precancerous lesions, liver cancer, and even cancer treatment, which provides clues for finding biomarkers of liver cancer, and different rats will have different disease progression due to individual differences, This suggests that we need to pay more attention to the individualization and accuracy of detection in the future. We can observe the changes in the immune system from the urine proteome in the early stage, enrich some biological pathways related to the process of an immune response, and distinguish the experimental groups with different immune strengths and weaknesses, which provides new ideas and directions for the future research of the immune system, and even speed up the research and development of vaccines. |
参考文献总数: | 53 |
馆藏号: | 硕071000/22027 |
开放日期: | 2023-06-18 |