浏览全部资源
扫码关注微信
Nogo66蛋白复合物对慢性高眼压大鼠视网膜微环境的影响
Effect of protein complex Nogo66-FA on retinal microenvironment in rats with chronic intraocular hypertension
- 第三军医大学学报 2021年43卷第7期 页码：614-621
- 作者机构：
400042 重庆，陆军军医大学（第三军医大学）大坪医院眼科
- 作者简介：
[贺翔鸽，E-mail：he_xiangge@sina.com HE Xiangge, E-mail: he_xiangge@sina.com
- 基金信息：
DOI：10.16016/j.1000-5404.202010079
中图分类号： R339.146;R392.13;R774.6收稿：2020-10-15，
修回：2020-12-28，
纸质出版：2021-04-15
- 稿件说明：
移动端阅览
程茗, 贺翔鸽. Nogo66蛋白复合物对慢性高眼压大鼠视网膜微环境的影响[J]. 第三军医大学学报, 2021,43(7):614-621.
Ming CHENG, Xiangge HE. Effect of protein complex Nogo66-FA on retinal microenvironment in rats with chronic intraocular hypertension[J]. Journal of Third Military Medical University, 2021, 43(7): 614-621.
程茗, 贺翔鸽. Nogo66蛋白复合物对慢性高眼压大鼠视网膜微环境的影响[J]. 第三军医大学学报, 2021,43(7):614-621. DOI： 10.16016/j.1000-5404.202010079.
Ming CHENG, Xiangge HE. Effect of protein complex Nogo66-FA on retinal microenvironment in rats with chronic intraocular hypertension[J]. Journal of Third Military Medical University, 2021, 43(7): 614-621. DOI： 10.16016/j.1000-5404.202010079.
摘要
目的
2
探讨Nogo66蛋白复合物对慢性高眼压大鼠视网膜微环境的影响。
方法
2
用532-二极管激光制备慢性高眼压大鼠模型，每只实验大鼠右眼为模型眼，左眼为对照眼。将造模成功的18只大鼠分为两组，分别接种Nogo66蛋白复合物和卵清蛋白复合物（
n
=9），荧光金逆行标记视网膜神经节细胞（retinal ganglion cells，RGCs）计数，免疫荧光法及免疫组化法观察视网膜GAP43、CD3、脑源性神经营养因子（brain-derived neurotrophic factor，BDNF）、胶质细胞源性神经营养因子（glial cell line-derived neurotrophic factor，GDNF）表达，Western blot检测视网膜BDNF、GDNF蛋白表达。
结果
2
慢性高眼压大鼠RGCs计数显著低于对照组大鼠（
P
<
0.01），接种Nogo66-FA复合物后，RGCs数量有所增加，显著高于对照组（
P
<
0.05）；且视网膜节细胞层和外丛状层有大量GAP43和CD3表达，视网膜节细胞层、内丛状层和内核层大量表达BDNF和GNDF，视网膜BNDF、GDNF蛋白表达量显著高于对照组（
P
<
0.05）。
结论
2
接种Nogo66蛋白复合物后，慢性高眼压模型大鼠视网膜微环境改善，T细胞及多种神经营养因子表达上调，RGCs数量增加。
Abstract
Objective
2
To investigate the effect of protein complex
Nogo66 and Freund's adjuvant (FA)
on retinal microenvironment in rats with chronic ocular hypertension.
Methods
2
After the rat model of chronic high intraocular pressure was established on the right eye (the left eye served as control) with the aid of 532-diode laser
the 18 model rats were equally divided into Nogo66 and ovalbumin groups
receiving vaccination with corresponding agent of 100 μg (in 300 μL FA) on the 1st
7th and 14th days. The retinal ganglion cells (RGCs) were counted after retrograde labelling with fluorescent gold. The retinal expression of CD3
growth associated protein 43 (GAP43)
brain-derived neurotrophic factor (BDNF) and glial cell line-derived neurotrophic factor (GDNF) were observed by immunofluorescence assay and immunohistochemical staining
and the protein levels of retinal BDNF and GDNF were detected by Western blotting.
Results
2
The count of RGCs was significantly lower in the right eyes with chronic ocular hypertension than the left eyes (
P
<
0.01). Inoculation of Nogo66-FA complex resulted in significantly larger number of RGCs when compared with the ra
ts from the ovalbumin group (
P
<
0.05). In addition
GAP43 and CD3 were highly expressed in the ganglion cell layer and the outer plexus layer
and BDNF and GNDF in the ganglion cell layer
the inner plexus layer and the inner core layer. The protein levels of BNDF and GDNF in the retina were significantly higher in the Nogo66-FA group than the ovalbumin group (
P
<
0.05).
Conclusion
2
Inoculation of protein complex Nogo66-FA can improve the retinal microenvironment and the activation of T cells
up-regulate the expression of various neurotrophic factors
and increase the count of RGCs in the rat model of chronic ocular hypertension.
关键词
Keywords
references
SEKINE Y, LINDBORG J A, STRITTMATTER S M. A proteolytic C-terminal fragment of Nogo-A (reticulon-4A) is released in exosomes and potently inhibits axon regeneration[J]. J Biol Chem, 2020, 295(8): 2175-2183. DOI:10.1074/jbc.ra119.009896.
DE LA VEGA GALLARDO N, DITTMER M, DOMBROWSKI Y, et al . Regenerating CNS myelin: Emerging roles of regulatory T cells and CCN proteins[J ] . Neurochem Int, 2019, 130: 104349. DOI:10.1016/j.neuint.2018.11.024.
BOURQUIN C, VAN DER HAAR M E, ANZ D, et al . DNA vaccination efficiently induces antibodies to Nogo-A and does not exacerbate experimental autoimmune encephalomyelitis[J ] . Eur J Pharmacol, 2008, 588(1): 99-105. DOI:10.1016/j.ejphar. 2008.04.026.
SOMMER C, LEINDERS M, ÜÇEYLER N. Inflammation in the pathophysiology of neuropathic pain[J]. Pain, 2018, 159(3): 595-602. DOI:10.1097/j.pain.0000000000001122.
PENG W. Neuroprotective effects of G-CSF administration in microglia-mediated reactive T cell activation in vitro [J ] . Immunol Res, 2017, 65(4): 888-902. DOI:10.1007/s12026-017-8928-9.
LOMBARDI M, PAROLISI R, SCARONI F, et al . Detrimental and protective action of microglial extracellular vesicles on myelin lesions: astrocyte involvement in remyelination failure[J ] . Acta Neuropathol, 2019, 138(6): 987-1012. DOI:10.1007/ s00401-019-02049-1.
BUSTO R, SERNA J, PERIANES-CACHERO A, et al . Ellagic acid protects from myelin-associated sphingolipid loss in experimental autoimmune encephalomyelitis[J ] . Biochim et Biophys Acta BBA-Mol Cell Biol Lipids, 2018, 1863(9): 958-967. DOI:10.1016/j.bbalip.2018.05.009.
STADELMANN C, TIMMLER S, BARRANTES-FREER A, et al . Myelin in the central nervous system: structure, function, and pathology[J ] . Physiol Rev, 2019, 99(3): 1381-1431. DOI:10.1152/physrev. 00031.2018.
程茗, 肖虹, 贺翔鸽. Nogo66蛋白疫苗接种对大鼠免疫应答的影响[J]. 解放军医学杂志, 2009, 34(5): 630-632. DOI: 10.3579/j.pmj.2009.05.630.
CHENG M, XIAO H, HE X G. Effects of inoculating Nogo66 vaccine on immunoresponse of rats[J]. Med J PLA, 2009, 34(5): 630-632. DOI: 10.3579/j.pmj.2009.05.630.
谢琳, 贺翔鸽, 苏踊跃, 等. Nogo-66融合蛋白的原核表达及鉴定[J]. 免疫学杂志, 2005, 21(3): 256-258. DOI: 10.3969/j.issn. 1000-8861.2005.03.026.
XIE L, HE X G, SU Y Y, et al . Identification and prokaryotic expression of Nogo-66[J ] . Immunol J, 2005, 21(3): 256-258. DOI: 10.3969/j.issn.1000-8861.2005.03.026.
TU S, LI K, DING X, et al . Relationship between intraocular pressure and retinal nerve fibre thickness loss in a monkey model of chronic ocular hypertension[J ] . Eye (Lond), 2019, 33(12): 1833-1841. DOI:10.1038/s41433-019-0484-1.
LIU H H, BUI B V, NGUYEN C T O, et al . Chronic ocular hypertension induced by circumlimbal suture in rats[J ] . Invest Ophthalmol Vis Sci, 2015, 56(5): 2811. DOI:10.1167/iovs.14-16009.
LAKSHMANAN Y, WONG F S Y, ZUO B, et al . Posttreatment intervention with Lycium barbarum polysaccharides is neuroprotective in a rat model of chronic ocular hypertension[J ] . Invest Ophthalmol Vis Sci, 2019, 60(14): 4606-4618. DOI:10.1167/iovs.19-27886.
LIU H H, FLANAGAN J G. A mouse model of chronic ocular hypertension induced by circumlimbal suture[J]. Invest Ophthalmol Vis Sci, 2017, 58(1): 353. DOI:10.1167/iovs.16-20576.
SEKINE Y, LINDBORG J A, STRITTMATTER S M. A proteolytic C-terminal fragment of Nogo-A (reticulon-4A) is released in exosomes and potently inhibits axon regeneration[J]. J Biol Chem, 2020, 295(8): 2175-2183. DOI:10.1074/jbc.ra119.009896.
NARDO G, TROLESE M C, VERDERIO M, et al . Counteracting roles of MHCI and CD8 + T cells in the peripheral and central nervous system of ALS SOD1G93A mice[J ] . Mol Neurodegener, 2018, 13(1): 42. DOI:10.1186/s13024-018-0271-7..
MOCKUS T E, REN H M, SHWETANK, et al . To go or stay: the development, benefit, and detriment of tissue-resident memory CD8 T cells during central nervous system viral infections[J ] . Viruses, 2019, 11(9): 842. DOI:10.3390/v11090842.
SMOLDERS J, HEUTINCK K M, FRANSEN N L, et al . Tissue-resident memory T cells populate the human brain[J ] . Nat Commun, 2018, 9: 4593. DOI:10.1038/s41467-018-07053-9.
HAASE S, HAGHIKIA A, WILCK N, et al . Impacts of microbiom e metabolites on immune regulation and autoimmunity[J ] . Immunology, 2018, 154(2): 230-238. DOI:10.1111/imm.12933.
CIGNARELLA F, CANTONI C, GHEZZI L, et al . Intermittent fasting confers protection in CNS autoimmunity by altering the gut microbiota[J ] . Cell Metab, 2018, 27(6): 1222-1235.e6. DOI:10. 1016/j.cmet.2018.05.006.
VAN KAER L, POSTOAK J L, WANG C, et al . Innate, innate-like and adaptive lymphocytes in the pathogenesis of MS and EAE[J ] . Cell Mol Immunol, 2019, 16(6): 531-539. DOI:10.1038/s41423-019- 0221-5.
GAUBLOMME J T, YOSEF N, LEE Y, et al . Single-cell genomics unveils critical regulators of Th17 cell pathogenicity[J ] . Cell, 2015, 163(6): 1400-1412. DOI:10.1016/j.cell.2015.11.009.
GOLAN M, MAUSNER-FAINBERG K, IBRAHIM B, et al . Fingolimod increases brain-derived neurotrophic factor level secretion from circulating T cells of patients with multiple sclerosis[J ] . CNS Drugs, 2019, 33(12): 1229-1237. DOI:10.1007/s40263-019- 00675-7.
TAN Z, KANG T, ZHANG X, et al . Nerve growth factor prevents arsenic-induced toxicity in PC12 cell s through the AKT/GSK-3β/NFAT pathway[J ] . J Cell Physiol, 2019, 234(4): 4726-4738. DOI:10.1002/jcp. 27255.
DUBOVý P, KLUSÁKOVÁ I, HRADILOVÁ-SVÍŽENSKÁ I, et al . Expression of regeneration-associated proteins in primary sensory neurons and regenerating axons after nerve injury-an overview[J ] . Anat Rec (Hoboken), 2018, 301(10): 1618-1627. DOI:10.1002/ar.23843.
BURSOVA S, DUBOVY P, VLCKOVA-MORAVCOVA E, et al . Expression of growth-associated protein 43 in the skin nerve fibers of patients with type 2 diabetes mellitus[J ] . J Neurol Sci, 2012, 315(1/2): 60-63. DOI:10.1016/j.jns.2011.11.038.
ZHOU B, YU P P, LIN M Y, et al . Facilitation of axon regeneration by enhancing mitochondrial transport and rescuing energy deficits[J ] . J Cell Biol, 2016, 214(1): 103-119. DOI:10.1083/jcb.201605101.
0
浏览量
0
下载量
0
CSCD
- 网络PDF下载
- Meta-XML下载
- BibTeX下载
- Endnote下载
- RefMan 下载
- NoteExpress下载
- NoteFirst下载
关联资源
相关文章
相关作者
相关机构
- 技术支持由北京北大方正电子有限公司提供 京ICP备09064830号-19
京公网安备11010802024621 - 本系统建议在Chrome、 IE9+ 以上版本浏览器阅读本站内容，360浏览器请切换至极速模式
- Cookies帮助我们提供服务并提供个性化体验。使用本网站，即表示您同意我们使用Cookies
