Neuroprotective effect of Nogo-66 receptor silencing in preterm rats with brain injury caused by intrauterine infection

ZHANG Shi-Fa; ZHOU Yan; ZHANG Kai-Jing; LUAN Jia-Jie; QI Shi-Mei

doi:10.7499/j.issn.1008-8830.2016.10.024

PDF(4402 KB)

Chinese Journal of Contemporary Pediatrics ›› 2016, Vol. 18 ›› Issue (10) : 1035-1043. DOI: 10.7499/j.issn.1008-8830.2016.10.024

EXPERIMENTAL RESEARCH

Neuroprotective effect of Nogo-66 receptor silencing in preterm rats with brain injury caused by intrauterine infection

ZHANG Shi-Fa¹, ZHOU Yan¹, ZHANG Kai-Jing¹, LUAN Jia-Jie², QI Shi-Mei³

Author information +

History +

Abstract

Objective To investigate the effect of Nogo-66 receptor (NgR) silencing with specific small interfering RNA (siRNA) on brain injury repair in preterm rats with brain injury caused by intrauterine infection and related mechanism of action. Methods The pregnant Sprague-Dawley rats (with a gestational age of 15 days) were selected, and premature delivery was induced by RU486 or lipopolysaccharide (LPS). The preterm rats delivered by those treated with RU486 were selected as the control group. The preterm rats with brain injury caused by intrauterine infection induced by LPS were divided into model, empty vector, and NgR-siRNA groups, with 36 rats in each group. The rats in the control and model groups were given routine feeding only, and those in the empty vector and NgR-siRNA groups were given an injection of lentiviral empty vector or NgR-siRNA lentivirus via the lateral ventricle on postnatal day 1 (P1) and then fed routinely. On P3, P7, and P14, 8 rats in each group were randomly selected and sacrificed to harvest the brain tissue. RT-PCR was used to measure the mRNA expression of NgR. Western blot was used to to measure the protein expression of active RhoA. The immunofluorescence histochemistry was used to determine the degree of activation of microglial cells and the morphology of oligodendrocyte precursor cells (OPCs). Hematoxylin and eosin staining was used to observe the pathological changes in brain tissue. The behavioral score was evaluated on P30. Results On P3, the NgR-siRNA group had significantly lower mRNA expression of NgR and protein expression of active RhoA in brain tissue than the model and empty vector groups (P < 0.05). In each group, the mRNA expression of NgR was positively correlated with the protein expression of active RhoA (P < 0.05). The results of immunofluorescence histochemistry showed that on P3, the NgR-siRNA group had a significantly reduced fluorescence intensity of the microglial cells labeled with CD11b compared with the model and empty vector groups (P < 0.05). The OPCs labeled with O4 antibody in the four groups were mainly presented with tripolar cell morphology. The results of pathological examination showed a normal structure of white matter with clear staining in the periventriclar area in the control group, a loose structure of white matter with disorganized fibers and softening lesions in the model and empty vector groups, and a loose structure of white matter with slightly disorganized fibers, slight gliocyte proliferation, and no significant necrotic lesions in the NgR-siRNA group. As for the behavioral score, compared with the model and empty vector groups, the NgR-siRNA group had a higher score in the suspension test, a longer total activity distance, and greater mean velocity and number of squares crossed, as well as a shorter time of slope test and a shorter time and distance of activity in the central area (P < 0.05), while there were no significant differences in these parameters between the NgR-siRNA and control groups (P > 0.05). Conclusions NgR silencing with specific siRNA can effectively silence the expression of NgR in pertem rats with brain injury caused by interauterine infection and has a significant neuroprotective effect in brain injury repair.

Key words

Nogo-66 receptor / Small interfering RNA / Intrauterine infection / Premature delivery / Rats

Cite this article

EndNote

Ris (Procite)

Bibtex

Download Citations

ZHANG Shi-Fa, ZHOU Yan, ZHANG Kai-Jing, LUAN Jia-Jie, QI Shi-Mei. Neuroprotective effect of Nogo-66 receptor silencing in preterm rats with brain injury caused by intrauterine infection[J]. Chinese Journal of Contemporary Pediatrics. 2016, 18(10): 1035-1043 https://doi.org/10.7499/j.issn.1008-8830.2016.10.024

References

[1] de Vries LS, Benders MJ, Groenendaal F. Progress in neonatal neurology with a focus on neuroimaging in the preterm infant[J]. Neuropediatrics, 2015, 46(4): 234-241.
[2] Mwaniki MK, Atieno M, Lawn JE, et al. Long-term neurodevelopmental outcomes after intrauterine and neonatal insults: a systematic review[J]. Lancet, 2012, 379(9814): 445-452.
[3] Pugni L, Pietrasanta C, Acaia B, et al. Chorioamnionitis and neonatal outcome in preterm infants: a clinical overview[J]. J Matern Fetal Neonatal Med, 2016, 29(9): 1525-1529.
[4] Migale R, Herbert BR, Lee YS, et al. Specific lipopolysaccharide serotypes induce differential maternal and neonatal inflammatory responses in a murine model of preterm labor[J]. Am J Pathol, 2015, 185(9): 2390-2401.
[5] Volpe JJ, Kinney HC, Jensen FE, et al. The developing oligodendrocyte: key cellular target in brain injury in the premature infant[J]. Int J Dev Neurosci, 2011, 29(4): 423-440.
[6] Borrie SC, Baeumer BE, Bandtlow CE. The Nogo-66 receptor family in the intact and diseased CNS[J]. Cell Tissue Res, 2012, 349(1): 105-117.
[7] Yan J, Zhou X, Guo JJ, et al. Nogo-66 inhibits adhesion and migration of microglia via GTPase Rho pathway in vitro[J]. J Neurochem, 2012, 120(5): 721-731.
[8] Wills ZP, Mandel-Brehm C, Mardinly AR, et al. The nogo receptor family restricts synapse number in the developing hippocampus[J]. Neuron, 2012, 73(3): 466-481.
[9] Cao Y, Dong YX, Xu J, et al. Spatiotemporal expression of Nogo-66 receptor after focal cerebral ischemia[J]. Neural Regen Res, 2016, 11(1): 132-136.
[10] Israelsson C, Flygt J, Åstrand E, et al. Altered expression of myelin-associated inhibitors and their receptors after traumatic brain injury in the mouse[J]. Restor Neurol Neurosci, 2014, 32(5): 717-731.
[11] Yin HL, Wang YL, Li JF, et al. Effects of curcumin on hippocampal expression of NgR and axonal regeneration in Aβ-induced cognitive disorder rats[J]. Genet Mol Res, 2014, 13(1): 2039-2047.
[12] Pourabdolhossein F, Mozafari S, Morvan-Dubois G, et al. Nogo receptor inhibition enhances functional recovery following lysolecithin-induced demyelination in mouse optic chiasm[J]. PLoS One, 2014, 9(9): e106378.
[13] Burd I, Balakrishnan B, Kannan S. Models of fetal brain injury, intrauterine inflammation, and preterm birth[J]. Am J Reprod Immunol, 2012, 67(4): 287-294.
[14] Wilson MD. Animal models of cerebral palsy: hypoxic brain injury in the newborn[J]. Iran J Child Neurol, 2015, 9(2): 9-16.
[15] 陈光福, 张蕴芳, 龙琦, 等. 丰富环境干预促进缺氧缺血性脑损伤新生大鼠神经元细胞增殖和功能修复[J]. 中国当代儿科杂志, 2012, 14(2): 139-143.
[16] Burd I, Chai J, Gonzalez J, et al. Beyond white matter damage: fetal neuronal injury in a mouse model of preterm birth[J]. Am J Obstet Gynecol, 2009, 201(3): 279.e1-279.e8.
[17] Pernet V, Schwab ME. The role of Nogo-A in axonal plasticity, regrowth and repair[J]. Cell Tissue Res, 2012, 349(1): 97-104.
[18] Shen Y. Traffic lights for axon growth: proteoglycans and their neuronal receptors[J]. Neural Regen Res, 2014, 9(4): 356-361.
[19] McDonald CL, Bandtlow C, Reindl M. Targeting the Nogo receptor complex in diseases of the central nervous system[J]. Curr Med Chem, 2011, 18(2): 234-244.
[20] Schmandke A, Mosberger AC, Schmandke A, et al. The neurite growth inhibitory protein Nogo-A has diverse roles in adhesion and migration[J]. Cell Adh Migr, 2013, 7(6): 451-454.
[21] Baburamani AA, Supramaniam VG, Hagberg H, et al. Microglia toxicity in preterm brain injury[J]. Reprod Toxicol, 2014, 48: 106-112.
[22] Kleinsimlinghaus K, Marx R, Serdar M, et al. Strategies for repair of white matter: influence of osmolarity and microglia on proliferation and apoptosis of oligodendrocyte precursor cells in different basal culture media[J]. Front Cell Neurosci, 2013, 7: 277.
[23] Boudreau RL, Rodríguez-Lebrón E, Davidson BL. RNAi medicine for the brain: progresses and challenges[J]. Hum Mol Genet, 2011, 20(R1): R21-R27.
[24] Huang JY, Wang YX, Gu WL, et al. Expression and function of myelin-associated proteins and their common receptor NgR on oligodendrocyte progenitor cells[J]. Brain Res, 2012, 1437: 1-15.
[25] Pedraza CE, Taylor C, Pereira A, et al. Induction of oligodendrocyte differentiation and in vitro myelination by inhibition of rho-associated kinase[J]. ASN Neuro, 2014, 6(4): 1-17.
[26] Sypecka J, Sarnowska A. The neuroprotective effect exerted by oligodendroglial progenitors on ischemically impaired hippocampal cells[J]. Mol Neurobiol, 2014, 49(2): 685-701.
[27] Stankiewicz TR, Linseman DA. Rho family GTPases: key players in neuronal development, neuronal survival, and neurodegeneration[J]. Front Cell Neurosci, 2014, 8: 314.
[28] Sanno H, Shen X, Kuru N, et al. Control of postnatal apoptosis in the neocortex by RhoA-subfamily GTPases determines neuronal density[J]. J Neurosci, 2010, 30(12): 4221-4231.
[29] Petrinovic MM, Duncan CS, Bourikas D, et al. Neuronal Nogo-A regulates neurite fasciculation, branching and extension in the developing nervous system[J]. Development, 2010, 137(15): 2539-2550.
[30] Shehadah A, Chen J, Cui X, et al. Combination treatment of experimental stroke with Niaspan and Simvastatin, reduces axonal damage and improves functional outcome[J]. J Neurol Sci, 2010, 294(1-2): 107-111.