木尼热·阿布都西库,席艳玲.前颗粒蛋白在脑卒中后失语症患者外周血单核细胞中的表达[J].中华物理医学与康复杂志,2023,45(4):289-296
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| 前颗粒蛋白在脑卒中后失语症患者外周血单核细胞中的表达 |
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| DOI：10.3760/cma.j.issn.0254-1424.2023.04.001 |
| 中文关键词: 脑卒中后失语 外周血单核细胞 前颗粒蛋白 kappaB结合蛋白相关核因子 |
| 英文关键词: Post-stroke aphasia Peripheral blood Mononuclear cells Aphasia-related progranulin genes Nuclear factors Kappa-B-binding protein |
| 基金项目:浦东新区科技发展基金事业单位民生科研专项项目（PKJ2022-Y30）；上海浦东新区光明中医医院院级科技基金项目（GMZYXK-202201） |
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| 中文摘要: |
| 目的 探究脑卒中后失语症(PSA)患者外周血单核细胞(PBMC)中失语相关基因前颗粒蛋白（GRN）的表达情况。 方法 将对数生长期PC12细胞进行培养, 细胞密度达到30%～50%时将其分为非特异性干扰组 (基因对照组)和特异性干扰组(基因沉默组)。在PC12 细胞中沉默GRN的表达后采用高通量转录组测序（RNA-seq）技术分析2组基因可变剪接事件发生情况。选取PSA患者10例（患者组）和健康受试者10例（对照组），2组受试者均于入组当天进行首次采血，并于入组12 d经皮（患者组出院当天）进行第二次采血。采用实时荧光定量聚合酶链式反应（RT-qPCR）测定2组受试者外周血单核细胞（PBMC）中GRN的mRNA表达量的变化和kappaB结合蛋白相关核因子（NFRKB）的可变剪接事件发生情况。患者组接受常规言语治疗，于首次采血和第二次采血后即刻采用汉语失语成套测试（ABC）对其言语功能进行评估，并采用Pearson法分析GRN表达量的变化与ABC各项评分变化的相关性。 结果 PC12细胞沉默GRN后，基因沉默组GRN的表达量与基因对照组GRN的表达量相比，差异有统计学意义（P<0.01）。两个样本间，发生显著差异可变剪接的基因共有237个，其中5′端发生可变剪接事件的基因数量最多。基因沉默组NFRKB可变剪接事件的发生概率与基因对照组比较，差异有统计学意义（P<0.05）；基因沉默组NFRKB可变剪接事件的发生概率与基因对照组比较，差异有统计学意义（P<0.05）。PSA患者组首次采血血样PBMC中GRN的mRNA表达水平和健康组血样比较，差异有统计学意义（P<0.001）；患者组第二次采血血样PBMC中GRN的mRNA表达水平与其首次采血血样比较，差异有统计学意义（P<0.001）。患者组首次采血血样PBMC中NFRKB基因发生可变剪接事件的概率与健康组比较，差异有统计学意义（P<0.05）。患者组第二次采血血样PBMC中NFRKB基因发生可变剪接事件的概率与首次采血比较，差异有统计学意义（P<0.01）。PSA患者第二次评估时的ABC量表口语表达得分显著优于其首次评估，差异有统计学意义（P<0.05），且在ABC量表的口语表达部分中，PSA患者第二次评估时复述评分，显著高于其首次评估，差异有统计学意义（P<0.05）。对患者组首次血样和第二次血样PBMC中GRN表达量的差异与PSA患者ABC量表口语表达评分的差异进行相关性分析，结果发现，GRN表达量的变化与口语表达能力的恢复呈正相关。 结论 GRN可通过调控NFRKB的可变剪接来促进PSA患者言语功能的恢复。 |
| 英文摘要: |
| Objective To document the expression of aphasia-related progranulin gene (GRN) in mononuclear cells in the peripheral blood (PBMC) of patients with post-stroke aphasia (PSA). Methods PC12 cells at the logarithmic-growth stage were cultured and divided into a non-specific interference group (the gene control group) and a specific interference group (the gene silencing group) when the cell density reached 30 to 50%. After the expression of GRN was knocked down in the cells, the occurrence of variable splicing events was analyzed using high-throughput transcriptome sequencing (RNA-seq). Meanwhile, 10 PSA patients were selected into a patient group and 10 healthy counterparts were chosen as a control group. Blood was collected from both groups and real-time fluorescence quantitative polymerase chain reactions (RT-qPCR) were employed to determine any changes in GRN mRNA expression and the occurrence of variable splicing events in the nuclear factor related to kappa-B-binding protein (NFRKB) in their PBMCs. The patient group received conventional speech therapy, and immediately after their first and second blood collections their speech functioning was assessed using the Chinese Aphasia Battery (ABC). Pearson correlation coefficients were then computed relating the GRN expression and ABC scores. Results After knocking down GRN in the PC12 cells, the expression of GRN in the gene knockdown group was significantly different from that in the control group. There were 237 genes with significant differences in variable splicing between the two samples. The number of genes with variable splicing events at the 5′ end was the largest. There were also significant differences between the groups in the average occurrence of NFRKB variable splicing events. And significant diffe-rences were observed in the mRNA expression of GRN between the two blood collections from the patient group, as well as between the first collection from the patient group and the controls. The average oral expression score of the PSA patients improved significantly, particularly the retelling score. The changes in the GRN expression level were positively correlated with the recovery of oral expression ability. Conclusion GRN can promote the recovery of speech function in PSA patients by regulating the variable splicing of NFRKB. |
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