儿童下呼吸道感染样本IFITMs基因差异表达分析

发布时间：2018-05-12 07:00

本文选题：呼吸道感染 + 病毒入侵　；参考：《济南大学》2017年硕士论文

【摘要】：病毒性呼吸道感染是呼吸道感染的主要组成部分。干扰素诱导的跨膜家族参与细胞的多种活动。人IFITMs家族蛋白有5类分别为IFITM1、IFITM2、IFITM3、IFITM5和IFITM10,均位于11号染色体。IFITM家族可以分为三类,一类是与免疫相关的IFITM1、IFITM2和IFITM3,参与抵抗病毒入侵的细胞免疫阶段。第二类是骨限制性表达的蛋白IFITM5,与骨中钙盐的沉积有关。第三类IFITM10可能与水环境适应性有关。鼠IFITMs家族中还有IFITM6和IFITM7,除IFITM7位于16号染色体外,其余均位于7号染色体。IFITM家族调节细胞的增殖和促进细胞的同性粘附,它们在多种肿瘤中过表达,与肿瘤细胞的迁移、侵袭和增殖密切相关。IFITM1、IFITM2和IFITM3参与抵抗病毒的入侵。IFITMs定位于细胞膜表面。IFITM1抵抗丝状病毒最有效,IFITM3抵抗甲型流感最有效。IFITM3定位于细胞晚期内含体途径,依赖于IFITM3蛋白的膜内区域1介导的活动,并且通过N-末端和IFITM蛋白中的保守的细胞内环路共同协作,最大化的限制病毒的入侵。IFITMs家族在生命活动中具有重要作用,在生物体内发生的功能作用及其机制还需要进一步研究。目的:分析呼吸道感染样本中IFITMs的表达差异,探寻IFITMs蛋白与呼吸道病毒感染的关系;参考大鼠组织IFITMs基因表达,分析IFITMs表达谱系。方法:(1)对下呼吸道感染样本进行临床信息分类汇总以及呼吸道病毒感染汇总。(2)设计并筛选人类IFITMs引物,通过RT-qPCR对下呼吸道感染样本中IFITMsmRNA进行扩增,分析IFITMs的相对表达。(3)提取大鼠组织RNA,设计并筛选大鼠IFITMs引物,利用RT-qPCR分析大鼠32种组织IFITMs表达。结果:本研究对320例儿童下呼吸道感染样本进行分析。发现IFITM3在支气管肺炎与重症肺炎、轻症肺炎与重症肺炎分类中的表达差异有统计学意义。IFITM3对流感病毒的限制能力和IFITM1对冠状病毒的限制能力区别于其它病毒。从呼吸道感染患者的肺泡灌洗液中提取的核酸,发现了IFITM5在mRNA水平上有表达。大鼠32种组织IFITMs mRNA表达结果表明IFITM1、IFITM2和IFITM3在各组中的表达比较普遍。在IFITM5的表达图谱中发现除了在骨骼中有分布,如骨髓、皮肤、胰和平滑肌等组织中均有表达。IFITM6在眼睛中的相对表达量最高,IFITM2与IFITM7的有类似的表达趋势。IFITM3、IFITM5和IFITM10在平滑肌中表达量均明显高表达与骨骼肌。结论:IFITM3蛋白在呼吸道感染加重过程中的相对表达发生变化。不同的IFITMs成员对同一病毒的限制能力不同。IFITMs限制病毒入侵的机制需进一步研究。在大鼠组织IFITMs表达谱系中,发现IFITM5在骨骼中的表达存在物种差异。由于同源性的关系,IFITM2和IFITM7,IFITM5和IFITM10在组织的表达图谱中有很多相似的地方,但是其在组织中的功能尚需进一步研究。
[Abstract]:Viral respiratory tract infection is the main component of respiratory tract infection. Interferon-induced transmembrane families are involved in a variety of cellular activities. There are five types of human IFITMs family proteins, IFITM1, IFITM2, IFITM3, IFITM5 and IFITM10, which are located on chromosome 11. IFITM1, IFITM2 and IFITM3, which are immune-related, are involved in the cellular immunity against virus invasion. The second is bone restricted protein IFITM 5, which is related to the deposition of calcium salt in bone. The third kind of IFITM10 may be related to the adaptability of water environment. IFITM6 and IFITM7 are also found in the IFITMs family of mice. Except for IFITM7 on chromosome 16, the rest are located on chromosome 7. IFITM family regulates the proliferation of cells and promotes the same sex adhesion of cells. They are overexpressed in many kinds of tumors and are related to the migration of tumor cells. Invasion and proliferation are closely related to. IFITM1IFITM2 and IFITM3 are involved in resisting virus invasiveness. IFITMs are located on the surface of cell membrane. IFITM1 is the most effective anti-filamentous virus. IFITM3 is the most effective anti-influenza A virus. IFITM3 is located in the late stage of cell inclusion pathway. Dependent on the intramembrane domain 1 of IFITM3 protein, and through the cooperation of N- terminal and conserved intracellular loop in IFITM protein, the restriction of virus invasion. IFITMs family plays an important role in life activities. The function and mechanism of the function in organism need to be further studied. Aim: to analyze the difference of IFITMs expression in respiratory tract infection samples, to explore the relationship between IFITMs protein and respiratory virus infection, and to analyze the IFITMs expression lineage by referring to the expression of IFITMs gene in rat tissues. Methods the human IFITMs primers were designed and screened for the clinical information classification of lower respiratory tract infection samples and respiratory virus infection collection. The IFITMsmRNA was amplified by RT-qPCR in the lower respiratory tract infection samples. The relative expression of IFITMs in rat tissues was analyzed. The rat IFITMs primers were designed and screened. RT-qPCR was used to analyze the expression of IFITMs in 32 tissues of rats. Results: 320 children with lower respiratory tract infection were analyzed. It was found that there were significant differences in the expression of IFITM3 between bronchopneumonia and severe pneumonia, mild pneumonia and severe pneumonia. The limiting ability of IFITM3 to influenza virus and IFITM1 to coronavirus were different from those of other viruses. Nucleic acid extracted from alveolar lavage fluid of patients with respiratory tract infection showed that IFITM5 was expressed at mRNA level. The expression of IFITMs mRNA in 32 tissues of rats showed that IFITM1, IFITM2 and IFITM3 were more common in each group. In the IFITM5 expression profile, it was found that except in the bone, such as bone marrow, skin, The relative expression of IFITM6 in the eyes was the highest in pancreatic and smooth muscle tissues. IFITM2 and IFITM7 had similar expression trend. IFITM3, IFITM5 and IFITM10 were highly expressed in smooth muscle and skeletal muscle. Conclusion the relative expression of the fraction IFITM3 protein changes during the exacerbation of respiratory tract infection. Different IFITMs members have different limiting ability to the same virus. In the IFITMs expression lineage of rat tissues, species differences were found in the expression of IFITM5 in bone. There are many similarities between IFITM2 and IFITM7 IFITM5 and IFITM10 in the tissue expression profiles due to homology, but their functions in tissues need further study.
【学位授予单位】：济南大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R725.6

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