基于生物信息学筛选创伤后应激障碍差异表达基因及验证  

To screen and verify the differentially expressed genes in post-traumatic stress disorder based on bioinformatics

作　　者：姜俊秀 邢文龙 徐婧姗 李吉鑫 张桂青[2] JIANG Junxiu;XING Wenlong;XU Jingshan;LI Jixin;ZHANG Guiqing(School of Medicine,Shihezi University,Shihezi,Xinjiang 832000,China;The First Affiliated Hospital of Shihezi University,Shihezi,Xinjiang 832000,China)

机构地区：[1]石河子大学医学院,新疆石河子832000 [2]石河子大学第一附属医院,新疆石河子832000

出　　处：《石河子大学学报（自然科学版）》2024年第6期734-741,共8页Journal of Shihezi University(Natural Science)

基　　金：国家自然科学基金项目(32260208)。

摘　　要：目的 筛选创伤后应激障碍(PTSD)中的差异表达基因(DEG)并进行验证,进一步探讨PTSD可能发生的机制。方法 分别收集PTSD患者与健康人群的外周血单个核细胞(PBMC)各26例,根据实验设计分别分为基因筛选组(n1=6)、基因验证组(n2=20)。首先通过转录组测序技术(RNA-Seq)建立新的基因库,然后使用GO分析、KEGG分析等方法筛选出PTSD患者的DEG,并进行实时荧光定量聚合酶链式反应(q-RT-PCR)验证;募集参与者的血浆样本,对上述DEG进行酶联免疫吸附测定(ELISA)进一步验证。结果 (1)筛选出DEG共609个,其中448个基因上调表达,161个基因下调表达。(2)q-RT-PCR验证基因4个,其中FPR1与CD14的表达显著升高,且差异具有统计学意义(P<0.05);XCL2与TIGIT的表达趋势呈下降,但差异不具有统计学意义(P<0.05)。(3)ELISA验证基因4个:其中CD14、FPR1、XCL2存在着蛋白水平的表达下调,差异具有统计学意义(P<0.05);而TIGIT的蛋白水平表达无明显差异,且不具有统计学意义。结论 CD14与FPR1为PTSD发生发展过程中的相关关键基因,其可能通过炎症通路来对PTSD的发生发展产生影响,而XCL2和TIGIT与PSTD发生发展的关系仍有待探究。Objective To screen and verify the differentially expressed genes(DEG)in post-traumatic stress disorder(PTSD),and further explore the possible mechanism of PTSD.Method Peripheral blood mononuclear cells(PBMC)were collected from 26 PTSD patients and 26 healthy people,respectively.According to the experimental design and technical requirements,they were divided into gene screening group(n1=6)and gene validation group(n2=20).First,the new gene pool was established by RNA sequencing technology(RNA-Seq),and then the DEG of PTSD patients were screened by GO analysis,KEGG analysis and other methods,and verified by real-time fluorescent quantitative polymerase chain reaction(q-RT-PCR).Plasma samples were collected from participants,and the DEG was further validated by enzyme-linked immunosorbent assay(ELISA).Results(1)A total of 609 DEG was screened out,of which 448 genes were up-regulated and 161 genes were down-regulated.(2)Four genes were verified by q-RT-PCR,among which the expression of FPR1 and CD14 was significantly increased,and the difference was statistically significant(P<0.05);The expression of XCL2 and TIGIT decreased,but the difference was not statistically significant.(3)Four genes were verified by ELISA:CD14,FPR1 and XCL2 were down-regulated at the protein level,and the difference was statistically significant(P<0.05);However,there was no significant difference in the protein expression of TIGIT,and it was not statistically significant(P<0.05).Conclusion CD14 and FPR1 for PTSD occurred in the development of differentially expressed genes,it might by inflammatory pathways to influence the occurrence of PTSD development.However,the relationship between XCL2 and TIGIT and the development of PSTD remains to be explored.

关 键 词：创伤后应激障碍 差异表达基因 CD14 FPR1 

分 类 号：R394[医药卫生—医学遗传学]