BACKGROUND:Nogo-neutralizing antibody IN-1 accelerates axon growth and enhances recovery of spinal cord function by inhibiting growth inhibitory factors. Neurotrophin-3 (NT-3)contributes to regeneration of nerve fiber...BACKGROUND:Nogo-neutralizing antibody IN-1 accelerates axon growth and enhances recovery of spinal cord function by inhibiting growth inhibitory factors. Neurotrophin-3 (NT-3)contributes to regeneration of nerve fibers in the spinal cord and motor function recovery. The combination of Nogo-neutralizing antibody IN-1 and NT-3 is hypothesized to produce better outcomes and facilitate axonal regeneration by affecting c-Fos and c-Jun protein expression. OBJECTIVE:To investigate the combined effects of Nogo-neutralizing antibody IN-1 and NT-3 on c-Fos and c-Jun protein levels in the injured spinal cord. DESIGN,TIME AND SETTING:A randomized,controlled study was performed at the Laboratory of Neuroanatomy,Xiangya Medical College,Central South University and the Central Laboratory of Third Xiangya Hospital of China from June 2005 to December 2007. MATERIALS:NT-3 (Peprotech,USA) and Nogo-neutralizing antibody IN-1 (Santa Cruz Biotechnology,USA) were used in this study. METHODS:Hemisectioned spinal cord injury models were established by cutting the posterior 2/3 of rat spinal cord,which is equivalent to the T8 level in the human spine. A total of 120 rats were equally and randomly assigned to three groups:model (0.2 μL saline),IN-1 (0.2 μL IN-1),and IN-1/NT-3 (0.2 μL IN-1 + 0.2 μL NT-3). The compounds were separately infused into transection sites on the side of head. MAIN OUTCOME MEASURES:Western blot analysis was employed to measure c-Fos and c-Jun protein expression in the injured spinal cord at 15,30 minutes,1,2,4,6,8,and 12 hours following surgery. RESULTS:Following spinal cord injury,c-Fos and c-Jun protein expression were increased and peaked at 4-6 hours. Following injection of IN-1 or the combination of IN-1 and NT-3,c-Fos protein expression was significantly reduced in the injured spinal cord (P < 0.05 or P < 0.01) (with the exception of the 15 minute time point). However,c-Jun protein expression was significantly increased (P < 0.05 or P < 0.01) (with the exception of the 15 and 30 minute time points). Combined application of IN-1 and NT-3 resulted in significantly altered protein expression compared to IN-1 alone. CONCLUSION:IN-1 increases c-Jun protein levels and protects the injured spinal cord by inhibiting c-Fos protein levels. Moreover,the effects of IN-1 combined with NT-3 are more significant than with IN-1 alone.展开更多
Neurotrophin-3 (NT3) is a growth factor found in many body tissues including the heart, intestines, skin, nervous system and in skeletal muscles including muscle spindles (Murase et al., 1994). NT3 is required for the...Neurotrophin-3 (NT3) is a growth factor found in many body tissues including the heart, intestines, skin, nervous system and in skeletal muscles including muscle spindles (Murase et al., 1994). NT3 is required for the survival, correct connectivity and function of sensory (“proprioceptive”) afferents that innervate muscle spindles;these neurons express receptors for NT3 including tropomyocin receptor kinase C. These proprioceptive afferents are important for normal movement (Boyce and Mendell, 2014) and signals from muscle spindles are important for recovery of limb movement (e.g., after spinal cord lateral hemisection)(Takeoka et al., 2014). The level of NT3 declines in most tissues during postnatal development;its level is low in adult and elderly humans and other mammals (Murase et al., 1994). Elevation of NT3 has been shown to improve outcome in various animal models of neurological disease and injury. For example, many groups have shown that delivery of NT3 directly into the central nervous system promotes recovery after spinal cord injury but this often involved invasive routes or gene therapy (Boyce and Mendell, 2014;Petrosyan et al., 2015;Wang et al., 2018).展开更多
Objective:To study the effects of neurotrophin-3 (NT-3) intervention on bone marrow mesenchymal stem cell osteoblast differentiation as well as cell proliferation and apoptosis. Methods: Bone marrow mesenchymal stem c...Objective:To study the effects of neurotrophin-3 (NT-3) intervention on bone marrow mesenchymal stem cell osteoblast differentiation as well as cell proliferation and apoptosis. Methods: Bone marrow mesenchymal stem cells were cultured and divided into control group, 25 ng/mL NT-3 group, 50 ng/mL NT-3 group and 100 ng/mL NT-3 group, they were treated with different doses of NT-3 for 24 h, and then osteoblast marker gene, cell proliferation gene and apoptosis gene expression were determined.Results: RUNX2, Osterix, ALP, OCN, BMP-2, Bcl-2, Nrf2, ERK1/2 and PCNA mRNA expression in 25 ng/mL NT-3 group, 50 ng/mL NT-3 group and 100 ng/mL NT-3 group were significantly higher than those in control group whereas Bim, Bax, Caspase-3, CHOP and Beclin1 mRNA expression were significantly lower than those in control group, and the larger the dose of NT-3, the higher the RUNX2, Osterix, ALP, OCN, BMP-2, Bcl-2, Nrf2, ERK1/2 and PCNA mRNA expression whereas the lower the Bim, Bax, Caspase-3, CHOP and Beclin1 mRNA expression.Conclusion: NT-3 intervention in bone marrow mesenchymal stem cells can promote osteoblast differentiation and cell proliferation and inhibit apoptosis.展开更多
基金a Grant from Department of Health of Hunan Province of China,No.B2005-076
文摘BACKGROUND:Nogo-neutralizing antibody IN-1 accelerates axon growth and enhances recovery of spinal cord function by inhibiting growth inhibitory factors. Neurotrophin-3 (NT-3)contributes to regeneration of nerve fibers in the spinal cord and motor function recovery. The combination of Nogo-neutralizing antibody IN-1 and NT-3 is hypothesized to produce better outcomes and facilitate axonal regeneration by affecting c-Fos and c-Jun protein expression. OBJECTIVE:To investigate the combined effects of Nogo-neutralizing antibody IN-1 and NT-3 on c-Fos and c-Jun protein levels in the injured spinal cord. DESIGN,TIME AND SETTING:A randomized,controlled study was performed at the Laboratory of Neuroanatomy,Xiangya Medical College,Central South University and the Central Laboratory of Third Xiangya Hospital of China from June 2005 to December 2007. MATERIALS:NT-3 (Peprotech,USA) and Nogo-neutralizing antibody IN-1 (Santa Cruz Biotechnology,USA) were used in this study. METHODS:Hemisectioned spinal cord injury models were established by cutting the posterior 2/3 of rat spinal cord,which is equivalent to the T8 level in the human spine. A total of 120 rats were equally and randomly assigned to three groups:model (0.2 μL saline),IN-1 (0.2 μL IN-1),and IN-1/NT-3 (0.2 μL IN-1 + 0.2 μL NT-3). The compounds were separately infused into transection sites on the side of head. MAIN OUTCOME MEASURES:Western blot analysis was employed to measure c-Fos and c-Jun protein expression in the injured spinal cord at 15,30 minutes,1,2,4,6,8,and 12 hours following surgery. RESULTS:Following spinal cord injury,c-Fos and c-Jun protein expression were increased and peaked at 4-6 hours. Following injection of IN-1 or the combination of IN-1 and NT-3,c-Fos protein expression was significantly reduced in the injured spinal cord (P < 0.05 or P < 0.01) (with the exception of the 15 minute time point). However,c-Jun protein expression was significantly increased (P < 0.05 or P < 0.01) (with the exception of the 15 and 30 minute time points). Combined application of IN-1 and NT-3 resulted in significantly altered protein expression compared to IN-1 alone. CONCLUSION:IN-1 increases c-Jun protein levels and protects the injured spinal cord by inhibiting c-Fos protein levels. Moreover,the effects of IN-1 combined with NT-3 are more significant than with IN-1 alone.
基金funded by the Brain Research Trust,the Rosetrees Trust and the International Spinal Research Trusta grant from the European Research Council under the European Union’s Seventh Framework Programme(FP/2007-2013)+5 种基金ERC Grant Agreement n.309731by a Research Councils UK Academic Fellowshipby the Medical Research Council(MRC)and the British Pharmacological Society(BPS)’s Integrative Pharmacology Fundsupported by the Dowager Countess Eleanor Peel Trusta Capacity Building Award in Integrative Mammalian Biology funded by the Biotechnology and Biological Sciences Research Council,BPSHigher Education Funding Council for England,Knowledge Transfer Partnerships,MRC and Scottish Funding Council
文摘Neurotrophin-3 (NT3) is a growth factor found in many body tissues including the heart, intestines, skin, nervous system and in skeletal muscles including muscle spindles (Murase et al., 1994). NT3 is required for the survival, correct connectivity and function of sensory (“proprioceptive”) afferents that innervate muscle spindles;these neurons express receptors for NT3 including tropomyocin receptor kinase C. These proprioceptive afferents are important for normal movement (Boyce and Mendell, 2014) and signals from muscle spindles are important for recovery of limb movement (e.g., after spinal cord lateral hemisection)(Takeoka et al., 2014). The level of NT3 declines in most tissues during postnatal development;its level is low in adult and elderly humans and other mammals (Murase et al., 1994). Elevation of NT3 has been shown to improve outcome in various animal models of neurological disease and injury. For example, many groups have shown that delivery of NT3 directly into the central nervous system promotes recovery after spinal cord injury but this often involved invasive routes or gene therapy (Boyce and Mendell, 2014;Petrosyan et al., 2015;Wang et al., 2018).
文摘Objective:To study the effects of neurotrophin-3 (NT-3) intervention on bone marrow mesenchymal stem cell osteoblast differentiation as well as cell proliferation and apoptosis. Methods: Bone marrow mesenchymal stem cells were cultured and divided into control group, 25 ng/mL NT-3 group, 50 ng/mL NT-3 group and 100 ng/mL NT-3 group, they were treated with different doses of NT-3 for 24 h, and then osteoblast marker gene, cell proliferation gene and apoptosis gene expression were determined.Results: RUNX2, Osterix, ALP, OCN, BMP-2, Bcl-2, Nrf2, ERK1/2 and PCNA mRNA expression in 25 ng/mL NT-3 group, 50 ng/mL NT-3 group and 100 ng/mL NT-3 group were significantly higher than those in control group whereas Bim, Bax, Caspase-3, CHOP and Beclin1 mRNA expression were significantly lower than those in control group, and the larger the dose of NT-3, the higher the RUNX2, Osterix, ALP, OCN, BMP-2, Bcl-2, Nrf2, ERK1/2 and PCNA mRNA expression whereas the lower the Bim, Bax, Caspase-3, CHOP and Beclin1 mRNA expression.Conclusion: NT-3 intervention in bone marrow mesenchymal stem cells can promote osteoblast differentiation and cell proliferation and inhibit apoptosis.
