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P1. Improvement of motor function induced by skeletal muscle contraction in spinal cord injury rats
The Spine Journal 19 (2019) S158−S194
ePosters: Basic Science/Biologics P1. Improvement of motor function induced by skeletal muscle contraction in spinal cord injury rats Norito Hayashi, MD1, Naoyuki Himi, PhD1, Emi Nakamura-Maruyama, PhD1, Naohiko Okabe, PhD, DVM2, Issei Sakamoto, MD1, Toru Hasegawa, MD, PhD1, Osamu Miyamoto, MD, PhD1; 1 Kawasaki Medical School Hospital, Matushima Kurashiki, Okayama, Japan; 2 Second Department of Physiology, Kawasaki Medical School, Kurashiki City, Okayama, Japan BACKGROUND CONTEXT: Previous reports have shown that treadmill training after spinal cord injury (SCI) contributes to functional recovery and is associated with increased levels of neurotrophic factors in the spinal cord in a rat model. However, the precise mechanism underlying the above observation has not been elucidated. Brain-derived neurotrophic factor (BDNF) is associated with motor function recovery due to its neuroprotective and axon sprouting effects. Promoting BDNF expression in spinal cord lesions by strategies such as physical training and cell transplantation has been reported to be effective in treating SCI in animals. Myokines are muscle-derived bioactive molecules associated with muscle contraction. Although BDNF is a myokine and considered a potential mediator of neuroplasticity following exercise, its contribution to motor function recovery after SCI has not yet been described in detail. PURPOSE: To investigate the role of muscle contraction in motor function recovery after SCI, with a focus on BDNF. STUDY DESIGN/SETTING: Eight-week-old Sprague-Dawley male rats were used to establish the SCI model. Percutaneous electrical muscle stimulation (10 mA, 2 Hz, 10 min) was applied to both limbs of the rats immediately after SCI. The stimulation was performed once per day for 4 weeks. The sham, SCI (SCI), and SCI with electrical muscle stimulation (SCIapt-ent-b 002B apt-ent-e + apt-ent-uES) groups were compared. OUTCOME MEASURES: The effect of electrical muscle stimulation was assessed by the motor function recovery. Histological assessments were done by measuring the cavity volume, the amount of axon sprouting, the number of apoptotic cells, and the amount of BDNF. METHODS: Spinal injury was induced by dropping a 20 g rod with a 2 mm apex diameter from a height of 25 mm onto the spine of an anesthetized rat at the T9 level. Motor function was assessed using the Basso-Beattie-Bresnahan Locomotor Scale (BBB score), the inclined plane test, and the Rotarod test. One week after the injury, transferase dUTP nick-end labeling (TUNEL)-positive cells were counted at the injury epicenter, and the level of BDNF was measured in both the spinal cord and the lower limb muscle. Four weeks after injury, the cavity volume of the epicenter and the level of phosphorylated growth-associated protein 43 (pGAP43) in the spinal cord were measured. RESULTS: A significantly improved BBB score and inclined plane test results were observed in the SCIapt-ent-b 002B apt-ent-e + apt-ent-uES group when compared to the SCI group, 4 weeks post-SCI. We also observed a decrease in the cavity volume and an increase in pGAP43 levels in the SCIapt-ent-b 002B apt-ent-e + apt-ent-uES group. Electrical muscle stimulation decreased the numbers of TUNEL-positive cells in the epicenter and increased the levels of BDNF in the spinal cord and lower limb muscles 1 week post-SCI. CONCLUSIONS: Electrical muscle stimulation improved motor function and increased BDNF levels in both the muscles and the spinal cords of rats
subjected to SCI. Muscle contraction-induced BDNF expression might be involved in motor recovery during rehabilitation. FDA DEVICE/DRUG STATUS: This abstract does not discuss or include any applicable devices or drugs. https://doi.org/10.1016/j.spinee.2019.05.425
P2. MiRNA-seq analysis of human vertebrae provides insight into the mechanism underlying GIOP Xiang Yu, PhD1, Hui Ren2, Jiang Xiaobing, PhD2, De Liang, MD3; 1 Guangzhou University of Chinese Medicine, Guangzhou, Baiyun/ Guangdong, China; 2 1st Affiliated Hospital of Guangzhou University of Chinese Medicine, Guang Zhou City, China; 3 Guangzhou, China BACKGROUND CONTEXT: High-throughput sequencing (HTS) was recently applied to detect microRNA (miRNA) regulation in age-related osteoporosis. However, miRNA regulation has not been reported in glucocorticoid-induced osteoporosis (GIOP) patients and the mechanism of GIOP remains elusive. PURPOSE: To comprehensively analyze the role of miRNA regulation in GIOP based on human vertebrae and to explore the molecular mechanism. STUDY DESIGN/SETTING: A high-throughput sequencing strategy was employed to identify miRNAs involved in GIOP. Twenty-six patients undergoing spinal surgery were included in this study. Six vertebral samples were selected for miRNA sequencing (miRNA-seq) analysis and 26 vertebral samples were verified by qRT-PCR. Bioinformatics was utilized for target prediction, to investigate the regulation of miRNA-mRNA networks, and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses. PATIENT SAMPLE: Twenty-six patients undergoing spinal surgery were included in this study. Six vertebral samples were selected for miRNA sequencing (miRNA-seq) analysis and 26 vertebral samples were verified by qRT-PCR. OUTCOME MEASURES: A high-throughput sequencing strategy was employed to identify miRNAs involved in GIOP. Twenty-six patients undergoing spinal surgery were included in this study. Six vertebral samples were selected for miRNA sequencing (miRNA-seq) analysis and 26 vertebral samples were verified by qRT-PCR. Bioinformatics was utilized for target prediction, to investigate the regulation of miRNA-mRNA networks, and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses. METHODS: A high-throughput sequencing strategy was employed to identify miRNAs involved in GIOP. Twenty-six patients undergoing spinal surgery were included in this study. Six vertebral samples were selected for miRNA sequencing (miRNA-seq) analysis and 6 vertebral samples were verified by qRT-PCR. Bioinformatics was utilized for target prediction, to investigate the regulation of miRNA-mRNA networks, and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses. RESULTS: Six significantly up-regulated miRNAs (including one novel miRNA) and three significantly down-regulated miRNAs were verified via miRNA-seq and verified in the vertebrae of GIOP patients. Up-regulated miRNAs included hsa-miR-214-5p, hsa-miR-10b-5p, hsa-miR-21-5p, hsamiR-451a, hsamiR-186-5p, and hsa-miR-novel-chr3_49,413 while downregulated miRNAs included hsa-let-7f-5p, hsa-let-7a-5p, and hsa-miR27a-3p. Bioinformatics analysis revealed 5,983 and 23,463 predicted targets in the up-regulated and down-regulated miRNAs respectively, using
Refer to onsite annual meeting presentations and postmeeting proceedings for possible referenced figures and tables. Authors are responsible for accurately reporting disclosure and FDA device/drug status at time of abstract submission.
ePosters: Basic Science/Biologics P1. Improvement of motor function induced by skeletal muscle contraction in spinal cord injury rats Norito Hayashi, MD1, Naoyuki Himi, PhD1, Emi Nakamura-Maruyama, PhD1, Naohiko Okabe, PhD, DVM2, Issei Sakamoto, MD1, Toru Hasegawa, MD, PhD1, Osamu Miyamoto, MD, PhD1; 1 Kawasaki Medical School Hospital, Matushima Kurashiki, Okayama, Japan; 2 Second Department of Physiology, Kawasaki Medical School, Kurashiki City, Okayama, Japan BACKGROUND CONTEXT: Previous reports have shown that treadmill training after spinal cord injury (SCI) contributes to functional recovery and is associated with increased levels of neurotrophic factors in the spinal cord in a rat model. However, the precise mechanism underlying the above observation has not been elucidated. Brain-derived neurotrophic factor (BDNF) is associated with motor function recovery due to its neuroprotective and axon sprouting effects. Promoting BDNF expression in spinal cord lesions by strategies such as physical training and cell transplantation has been reported to be effective in treating SCI in animals. Myokines are muscle-derived bioactive molecules associated with muscle contraction. Although BDNF is a myokine and considered a potential mediator of neuroplasticity following exercise, its contribution to motor function recovery after SCI has not yet been described in detail. PURPOSE: To investigate the role of muscle contraction in motor function recovery after SCI, with a focus on BDNF. STUDY DESIGN/SETTING: Eight-week-old Sprague-Dawley male rats were used to establish the SCI model. Percutaneous electrical muscle stimulation (10 mA, 2 Hz, 10 min) was applied to both limbs of the rats immediately after SCI. The stimulation was performed once per day for 4 weeks. The sham, SCI (SCI), and SCI with electrical muscle stimulation (SCIapt-ent-b 002B apt-ent-e + apt-ent-uES) groups were compared. OUTCOME MEASURES: The effect of electrical muscle stimulation was assessed by the motor function recovery. Histological assessments were done by measuring the cavity volume, the amount of axon sprouting, the number of apoptotic cells, and the amount of BDNF. METHODS: Spinal injury was induced by dropping a 20 g rod with a 2 mm apex diameter from a height of 25 mm onto the spine of an anesthetized rat at the T9 level. Motor function was assessed using the Basso-Beattie-Bresnahan Locomotor Scale (BBB score), the inclined plane test, and the Rotarod test. One week after the injury, transferase dUTP nick-end labeling (TUNEL)-positive cells were counted at the injury epicenter, and the level of BDNF was measured in both the spinal cord and the lower limb muscle. Four weeks after injury, the cavity volume of the epicenter and the level of phosphorylated growth-associated protein 43 (pGAP43) in the spinal cord were measured. RESULTS: A significantly improved BBB score and inclined plane test results were observed in the SCIapt-ent-b 002B apt-ent-e + apt-ent-uES group when compared to the SCI group, 4 weeks post-SCI. We also observed a decrease in the cavity volume and an increase in pGAP43 levels in the SCIapt-ent-b 002B apt-ent-e + apt-ent-uES group. Electrical muscle stimulation decreased the numbers of TUNEL-positive cells in the epicenter and increased the levels of BDNF in the spinal cord and lower limb muscles 1 week post-SCI. CONCLUSIONS: Electrical muscle stimulation improved motor function and increased BDNF levels in both the muscles and the spinal cords of rats
subjected to SCI. Muscle contraction-induced BDNF expression might be involved in motor recovery during rehabilitation. FDA DEVICE/DRUG STATUS: This abstract does not discuss or include any applicable devices or drugs. https://doi.org/10.1016/j.spinee.2019.05.425
P2. MiRNA-seq analysis of human vertebrae provides insight into the mechanism underlying GIOP Xiang Yu, PhD1, Hui Ren2, Jiang Xiaobing, PhD2, De Liang, MD3; 1 Guangzhou University of Chinese Medicine, Guangzhou, Baiyun/ Guangdong, China; 2 1st Affiliated Hospital of Guangzhou University of Chinese Medicine, Guang Zhou City, China; 3 Guangzhou, China BACKGROUND CONTEXT: High-throughput sequencing (HTS) was recently applied to detect microRNA (miRNA) regulation in age-related osteoporosis. However, miRNA regulation has not been reported in glucocorticoid-induced osteoporosis (GIOP) patients and the mechanism of GIOP remains elusive. PURPOSE: To comprehensively analyze the role of miRNA regulation in GIOP based on human vertebrae and to explore the molecular mechanism. STUDY DESIGN/SETTING: A high-throughput sequencing strategy was employed to identify miRNAs involved in GIOP. Twenty-six patients undergoing spinal surgery were included in this study. Six vertebral samples were selected for miRNA sequencing (miRNA-seq) analysis and 26 vertebral samples were verified by qRT-PCR. Bioinformatics was utilized for target prediction, to investigate the regulation of miRNA-mRNA networks, and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses. PATIENT SAMPLE: Twenty-six patients undergoing spinal surgery were included in this study. Six vertebral samples were selected for miRNA sequencing (miRNA-seq) analysis and 26 vertebral samples were verified by qRT-PCR. OUTCOME MEASURES: A high-throughput sequencing strategy was employed to identify miRNAs involved in GIOP. Twenty-six patients undergoing spinal surgery were included in this study. Six vertebral samples were selected for miRNA sequencing (miRNA-seq) analysis and 26 vertebral samples were verified by qRT-PCR. Bioinformatics was utilized for target prediction, to investigate the regulation of miRNA-mRNA networks, and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses. METHODS: A high-throughput sequencing strategy was employed to identify miRNAs involved in GIOP. Twenty-six patients undergoing spinal surgery were included in this study. Six vertebral samples were selected for miRNA sequencing (miRNA-seq) analysis and 6 vertebral samples were verified by qRT-PCR. Bioinformatics was utilized for target prediction, to investigate the regulation of miRNA-mRNA networks, and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses. RESULTS: Six significantly up-regulated miRNAs (including one novel miRNA) and three significantly down-regulated miRNAs were verified via miRNA-seq and verified in the vertebrae of GIOP patients. Up-regulated miRNAs included hsa-miR-214-5p, hsa-miR-10b-5p, hsa-miR-21-5p, hsamiR-451a, hsamiR-186-5p, and hsa-miR-novel-chr3_49,413 while downregulated miRNAs included hsa-let-7f-5p, hsa-let-7a-5p, and hsa-miR27a-3p. Bioinformatics analysis revealed 5,983 and 23,463 predicted targets in the up-regulated and down-regulated miRNAs respectively, using
Refer to onsite annual meeting presentations and postmeeting proceedings for possible referenced figures and tables. Authors are responsible for accurately reporting disclosure and FDA device/drug status at time of abstract submission.