急性肝功能衰竭相关基因和蛋白的研究

发布时间：2018-06-05 16:47

本文选题：急性肝功能衰竭 + 四氯化碳　；参考：《河南师范大学》2016年硕士论文

【摘要】：急性肝功能衰竭(或急性肝衰竭,acute hepatic failure,AHF)是多种诱因引起的、肝脏细胞受损严重并致使其多种功能发生障碍的病死率极高和对人们的健康威胁较大的一种肝脏疾病,常伴随多种器官衰竭、肝性脑病、血液动力学紊乱和多种代谢疾病等并发症,因其发生机制复杂和影响因素多等原因,致使目前对其发病机理的研究虽然较多但仍尚不完全清楚,且仍没有发现对其有效的防御和治疗办法,目前仍以综合治疗为主,故而一直以来急性肝衰竭都是临床医学上最具挑战的疾病之一。为此,本文成功构建了大鼠AHF模型,并通过检测其血清ALT和AST两种酶活性、计算其肝系数和冰冻切片显微镜检检验建模效果;Rat Genome 230 2.0基因芯片检测其基因表达变化发现了4819个AHF相关基因,IPA分析其相关基因表达变化发现,IL-1、IL-6和IL-8等信号通路信号传导活动增强,TP53、ATM和AMPK等减弱;细胞存活、增殖和分化等生理活动增强,T/B淋巴细胞凋亡减弱;分析AHF发生机理相关基因表达变化后预测,通过IL-1α→IL-1R1→→MAPK8→FOS/JUN和/或TNFα→TNFRSF1A/B→→Caspases途径在AHF损伤和进展阶段调节细胞凋亡、炎症和免疫反应,炎症因子通过NF-κB途径在进展阶段调节细胞凋亡,TP53在进展阶段抑制细胞凋亡,恢复阶段细胞凋亡被抑制(此时Caspase3表达下调),进展和恢复阶段通过肝细胞增殖来修复其损伤(PCNA表达上调),因此,本文推测,在CCl4诱导的急性肝衰竭中,IL-1R1、TNFRSF1A/B、Caspase3、TP53、PCNA和NF-κB等基因/蛋白起重要作用。本文选择使用本实验室所创的液压转基因技术(hydrodynamics-based transgene,HDT)对小鼠AHF进行基因治疗初探,通过血清ALT和AST和冰冻切片显微检测鉴定小鼠AHF模型建立成功,通过不同注射方式条件摸索(液压转基因注射生理盐水/空载体质粒-生理盐水溶液与灌胃建模的前后和不同时间差)发现,灌胃建模后6h,液压转基因注射空载体质粒pEGFP-C1与液压转基因注射生理盐水相比,对小鼠AHF形成的影响结果无显著性差异,从而为AHF的基因治疗打下一定基础。综上所述,本文进一步阐述和分析了AHF的发生机理,探索了液压转基因目的基因进入AHF模型体内的最佳注射条件,为更进一步的研究及其治疗奠定基础。
[Abstract]:Acute hepatic failure (AHF) is caused by many inducements. Often accompanied by multiple organ failure, hepatic encephalopathy, hemodynamic disorders and many metabolic diseases and other complications, because of its complex mechanism and influence factors and other reasons, As a result, although there are many studies on its pathogenesis at present, it is still not fully understood, and no effective defense and treatment methods have been found. At present, comprehensive treatment is still the main treatment. Acute liver failure has always been one of the most challenging diseases in clinical medicine. Therefore, the rat AHF model was successfully constructed, and the activities of serum ALT and AST were detected. The liver coefficient was calculated and the model was examined by frozen section microscope. The gene expression changes of Rat Genome 230 2.0 gene microarray were detected. The expression changes of 4819 AHF related genes were analyzed and the signal pathways such as IL 1, IL 6 6 and IL-8 were found. The signal transduction activity was enhanced, and TP53 / ATM and AMPK were weakened. Cell survival, proliferation, differentiation and other physiological activities enhanced apoptosis of T / B lymphocytes. Apoptosis, inflammation and immune response were regulated by IL-1 伪 IL-1R1 MAPK8 FOS/JUN and / or TNF 伪 TNFRSF1A/B Caspases pathway during AHF injury and progression. Inflammatory factors regulate apoptosis through NF- 魏 B pathway. TP53 inhibits apoptosis at the progressive stage. Apoptosis was inhibited during the recovery phase (the expression of Caspase3 was down-regulated at this time, and the expression of Caspase3 was up-regulated by the proliferation of hepatocytes during the recovery phase. In acute hepatic failure induced by CCl4, IL-1R1 / TNFRSF1 / TNFRSF1 / TNFRSF1 / TNFRSF1 / TNFRSF1 / TNFRSF1 / TNFRSF1 / TNFRSF1 / TP53 / protein play an important role in acute hepatic failure induced by CCl4. In this paper, hydrodynamics-based transgeneic gene therapy was used to treat AHF in mice. Serum ALT, AST and frozen section microanalysis were used to identify the successful establishment of mouse AHF model. It was found that different injection conditions (hydraulic transgenic injection of normal saline / no-load granulus-saline solution and the difference of time before and after gastric perfusion), There was no significant difference in the effect of hydraulic transgenic injection of no-load body mass pEGFP-C1 and hydraulic transgenic injection of normal saline on the formation of AHF in mice at 6 h after gastric perfusion, which laid a foundation for the gene therapy of AHF. To sum up, the mechanism of AHF was further expounded and analyzed, and the optimal injection conditions of hydraulic transgenic gene into AHF model were explored, which laid a foundation for further research and treatment.
【学位授予单位】：河南师范大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R575.3

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