title: [Overexpression of miR-125b promotes apoptosis of macrophages].
abstract: Objective To investigate the expressions of miR-125b and target gene Raf1 proto-oncogene serine/threonine protein kinase (RAF1) in peripheral blood mononuclear cells (PBMCs) of pediatric patients with pulmonary tuberculosis (PTB), and observe the regulation of miR-125b on macrophage apoptosis and activity. Methods PBMCs of patients with PTB and healthy children were collected and separated. Real-time fluorescence quantitative PCR was used to detect mRNA expression level of miR-125b and RAF1, and Western blotting was used to detect the protein level of RAF1. THP-1 macrophages were transfected into miR-125b mimic, negative control mimic (NC-mimic), miR-125b inhibitor and negative control inhibitor (NC-inhibitor), which were cultured for 48 hours. Western blotting was performed to observe the expression of RAF1 in THP-1 macrophages, annexin V-FITC/PI double staining combined with flow cytometry was used to test cell apoptosis, and CCK-8 assay was used to detect cell proliferation. Results The expression of miR-125b in PBMCs in pediatric patients with PTB was downregulated, and mRNA and protein levels of RAF1 were upregulated. When miR-125b was over-expressed in THP-1 macrophages, the expression of RAF1 was reduced to promote the apoptosis of macrophages and decrease cell activity; when the expression of miR-125b was inhibited in THP-1 macrophages, the expression of RAF1 was elevatedand the apoptosis of macrophages was inhibited, the cell activity was promoted. Conclusion In PBMCs of children with PTB, miR-125b level is low. Upregulation of miR-125b in THP-1 macrophages, the apoptosis of THP-1 macrophages is promoted and cell activity is inhibited.
authors.affiliation: Department of Gastroenterology, Children's Hospital of Chongqing Medical University, Chongqing 400014, China.
authors.forename: Guangyuan
authors.initials: G
authors.lastname: Yu
authors.affiliation: Department of Gastroenterology, Children's Hospital of Chongqing Medical University, Chongqing 400014, China. *Corresponding authors, E-mail: zhanxue@hotmail.com.
authors.forename: Xue
authors.initials: X
authors.lastname: Zhan
authors.affiliation: Department of Infection, Children's Hospital of Chongqing Medical University, Chongqing 400014, China. *Corresponding authors, E-mail: 39003614@qq.com.
authors.forename: Zhenzhen
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: Pediatrics Research Institute, Children's Hospital of Chongqing Medical University, Ministry-of-Education Key Laboratory of Child Development and Disorders, China International Science and Technology Cooperation Base of Child Development and Critical Disorders, Chongqing Key Laboratory of Pediatrics, Chongqing 400014, China.
authors.forename: Yasha
authors.initials: Y
authors.lastname: Li
mesh_terms: D000293:Adolescent; D017209:Apoptosis / Q000235:genetics*; D015153:Blotting, Western; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D002648:Child; D002675:Child, Preschool; D005260:Female; D005434:Flow Cytometry; D015870:Gene Expression; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D007963:Leukocytes, Mononuclear / Q000378:metabolism; D008264:Macrophages / Q000378:metabolism*; D008297:Male; D035683:MicroRNAs / Q000235:genetics*; D000090063:Proto-Oncogene Mas; D019908:Proto-Oncogene Proteins c-raf / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014397:Tuberculosis, Pulmonary / Q000097:blood / Q000235:genetics / Q000378:metabolism
chemical_list: C000715470:MAS1 protein, human; C531888:MIRN125 microRNA, human; D035683:MicroRNAs; D000090063:Proto-Oncogene Mas; D019908:Proto-Oncogene Proteins c-raf