title: MiR-139-5p inhibits HGTD-P and regulates neuronal apoptosis induced by hypoxia-ischemia in neonatal rats.
abstract: Human growth transformation dependent protein (HGTD-P) is a newly identified protein that promotes neuronal apoptosis in hypoxia-ischemia brain damage (HIBD) in neonatal rats. However, the mechanisms regulating HGTD-P expression are not clear. Here we describe microRNAs targeted to HGTD-P and examine their effects on regulating neuronal apoptosis in HIBD. We use samples from cultured neurons after oxygen-glucose deprivation (OGD) and postnatal day 10 rat brains after hypoxia-ischemia (HI). RT-PCR, Western blotting, and immunostaining are used to detect the expression of HGTD-P and cleaved caspase 3, as well as real-time PCR detects microRNA expression. MicroRNA agomir is used to inhibit the expression of HGTD-P, and DAPI, TUNEL, and TTC staining are employed to detect cell apoptosis and brain damage. Moreover, in vitro processing assay is used to examine the mechanism by which HI down-regulates miR-139-5p expression. We found that miR-139-5p is down-regulated in neurons and rat brains after HI treatment. The expression pattern of miR-139-5p correlates inversely with that of HGTD-P. Furthermore, miR-139-5p agomir inhibits neuronal apoptosis and attenuates HIBD, which is concurrent with down-regulation of HGTD-P. Moreover, pre-miR-139 processing activity decreases in extracts from OGD neurons, and OGD neuronal extracts attenuates the processing of pre-miR-139 by Dicer. In conclusion, HI induces inhibitors which block the processing step of pre-miR-139, resulting in the down-regulation of mature miR-139-5p. The down-regulation of miR-139-5p plays a critical role in the up-regulation of HGTD-P expression. MiR-139-5p agomir attenuates brain damage when used 12h after HI, providing a longer therapeutic window than anti-apoptosis compounds currently available.
authors: Department of Pediatrics, West China Second University Hospital, Sichuan University, Chengdu 610041, China; Key Laboratory of Obstetric & Gynecologic and Pediatric Diseases and Birth Defects of Ministry of Education, Sichuan University, 610041 Chengdu, Sichuan, China. Electronic address: quyi712002@163.com.
authors: Yi
authors: Y
authors: Qu
authors: Department of Pediatrics, West China Second University Hospital, Sichuan University, Chengdu 610041, China.
authors: Jinlin
authors: J
authors: Wu
authors: Department of Pediatrics, West China Second University Hospital, Sichuan University, Chengdu 610041, China.
authors: Dapeng
authors: D
authors: Chen
authors: Department of Pediatrics, West China Second University Hospital, Sichuan University, Chengdu 610041, China; Key Laboratory of Obstetric & Gynecologic and Pediatric Diseases and Birth Defects of Ministry of Education, Sichuan University, 610041 Chengdu, Sichuan, China.
authors: Fengyan
authors: F
authors: Zhao
authors: Department of Pediatrics, West China Second University Hospital, Sichuan University, Chengdu 610041, China; Key Laboratory of Obstetric & Gynecologic and Pediatric Diseases and Birth Defects of Ministry of Education, Sichuan University, 610041 Chengdu, Sichuan, China.
authors: Junyan
authors: J
authors: Liu
authors: West China Medical Center, Sichuan University, 610041 Chengdu, Sichuan, China.
authors: Chunlei
authors: C
authors: Yang
authors: West China Medical Center, Sichuan University, 610041 Chengdu, Sichuan, China.
authors: Dapeng
authors: D
authors: Wei
authors: Department of Pediatrics and Neurology, University of California, San Francisco, CA 94143, USA.
authors: Donna M
authors: DM
authors: Ferriero
authors: Department of Pediatrics, West China Second University Hospital, Sichuan University, Chengdu 610041, China; Key Laboratory of Obstetric & Gynecologic and Pediatric Diseases and Birth Defects of Ministry of Education, Sichuan University, 610041 Chengdu, Sichuan, China; Department of Pediatrics and Neurology, University of California, San Francisco, CA 94143, USA. Electronic address: mudz@scu.edu.cn.
authors: Dezhi
authors: D
authors: Mu
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D017209:Apoptosis / Q000187:drug effects / Q000528:radiation effects*; D020520:Brain Infarction / Q000209:etiology / Q000473:pathology; D002478:Cells, Cultured; D004195:Disease Models, Animal; D004622:Embryo, Mammalian; D005786:Gene Expression Regulation / Q000235:genetics / Q000502:physiology*; D005947:Glucose / Q000172:deficiency; D006801:Humans; D000860:Hypoxia / Q000378:metabolism; D020925:Hypoxia-Ischemia, Brain / Q000473:pathology*; D008565:Membrane Proteins / Q000378:metabolism*; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D024101:Mitochondrial Proteins / Q000378:metabolism*; D060172:Morpholinos / Q000494:pharmacology; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D010751:Phosphopyruvate Hydratase / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D013997:Time Factors
chemical_list: C486647:FAM162A protein, human; C555098:MIRN139 microRNA, human; D008565:Membrane Proteins; D035683:MicroRNAs; D024101:Mitochondrial Proteins; D060172:Morpholinos; D010751:Phosphopyruvate Hydratase; D005947:Glucose
keywords: 3′-untranslated regions; 3′UTR; AI; AIF; Apoptosis; Brain; CC3; CCA; HGTD-P; HI; HIBD; Hypoxia–ischemia; MicroRNA; OGD; RISC; RNA-induced silencing complex; TTC; apoptosis inducible factor; apoptotic index; cleaved caspase 3; common carotid artery; human growth transformation dependent protein; hypoxia–ischemia; hypoxia–ischemia brain damage; oxygen–glucose deprivation; tPA; tissue plasminogen activator; triphenyltetrazolium chloride