荧光金逆行标记评价视神经切断后视网膜节细胞存活率

王能; 李茂; 栾文斌; 项平

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荧光金逆行标记评价视神经切断后视网膜节细胞存活率

王能 , 李茂 , 栾文斌 , 项平

文章导航 > 蚌埠医学院学报 > 2014, 38 > 2: (148-160)

引用本文:

Citation:

荧光金逆行标记评价视神经切断后视网膜节细胞存活率

1.
1. 蚌埠医学院第一附属医院中心实验室,
2.
3.
2. 组织移植安徽省重点实验室, 安徽蚌埠 233004,
4.
5.
3. 山东省蓬莱市中医院眼耳鼻喉科, 265600

作者简介: 王能(1984- ),男，硕士研究生.

基金项目:
安徽省教育厅自然科学研究重点项目(KJ2010A241)

Evaluating the survival rate of axotomized retinal ganglion cells by fluorogold retrograde labeling

1.
1. Department of Central Laboratory, The First Affiliated Hospital of Bengbu Medical College, Bengbu Anhui 233004;
2.
2. Department of Key Laboratory of Tissue Transplantation, Bengbu Anhui 233004;
3.
3. Department of Otolaryngology, Penglai Traditional Chinese Medicine Hospital, Penglai Shandong 265600, China

摘要: 目的：利用荧光金逆行标记评价视神经切断后视网膜节细胞(RGCs)的存活率。方法：将SD大鼠20只随机均分为4组,正常对照组和视神经切断后5 d、7 d和14 d组。用荧光金逆行标记和定量解剖学技术观察正常对照组和视神经切断后5 d、7 d和14 d组RGCs的密度。结果:正常对照组RGCs呈放射状分布,边界清晰,可见明显细胞突起,无渗漏现象;视神经切断组RGCs随时间延长而减少,细胞分布不均匀,并且可见吞噬死亡细胞碎片和从死亡细胞渗漏的荧光金的小胶质细胞。正常对照组左眼与右眼RGCs密度差异无统计学意义(P0.05);视神经切断后RGCs进行性减少,切断后5 d、7 d和14 d组左眼RGCs密度均明显低于正常对照组(P0.01),视神经切断后5 d、7 d和14 d存活率分别为53%、38%和13%。结论：荧光金逆行标记是评价视神经切断后RGCs存活率有效的方法。
- 视神经切断 /
- 荧光金 /
- 视网膜节细胞
Abstract: Objective: To evaluate the survival rate of retinal ganglion cells(RGCs) after transsection of the optic nerve using the method of fluorogold retrograde labeling. Methods: Twenty SD rats were randomly divided inObjective: To four groups: normal control group，5 d，7 d and 14 d after transsection of the optic nerve groups. The RGCs density of the normal control group and 5 d，7 d and 14 d after transsection of the optic nerve groups was observed by fluorogold retrograde labeling and quantitative anaObjective: To mical technology. Results: In the normal control group， the RGCs labeled by the fluorescent gold presented radial distribution，clear boundary and obvious cell processes without any leak. In the optic nerve transsection groups， the number of RGCs decreased with time increasing and the cell distribution was not uniform; and the debris of dead cells and cell leakage from the dead FG microglial cells were observed. In thenormal control group， the average density of RGCs of the left eye had no significant difference compared Objective: To that of the right eye(P ＞0. 05). In the 5 d，7 d and 14 d after transsection of the optic nerve groups， the number of RGCs was reduced progressively and the density of RGCs of the left eye was significantly lower than that in the normal control group(P ＜ 0. 01) ; the survival rates of RGCs in the groups of 5 d，7 d and 14 d after optic nerve transsection were 53%，38% and 13%，respectively. Conclusions: luorogold retrograde labeling is an effective method of evaluating the survival rate of axoObjective: Tomized retinal ganglion cells.
- transection of the optic nerve /
- fluorescent gold /
- retinal ganglion cells

[2] 徐建国.慢性疼痛的药物治疗进展[J].医学研究生学报，2007，20(2) : 113 - 119.
[2]	李小梅，刘瑞琪，李红英，等.恶性肿瘤患者临终前的阿片类药物止痛回顾[J].中国肿瘤临床，2005，32(23) : 1358 - 1363.
[3] Cleeland CS，Gonin R，Hatfield AK， et al.Pain and its treatment in out patients with metastatic cancer[J].N Engl J Med，1994，330(9) : 592 - 596.
[4] Lotsch J.Opioid metabolites[J].J Pain Symptom Anage，2005，29(5 Suppl) : S10 - S24.
[5] Jasinski DR，Preston KL.Evaluation of tilidine for morphine-like subjects and euphoria[J].Drug Alcohol Depend，1986，18 (3) :273 - 292.
[6] Thierry C，Boeynaems JM，Paolo M.Actions of tilidine and nortilidine on cloned opioid receptors[J].Eur J Pharmacol，2005，506(3) : 205 - 208.
[7] 陈素青，黄新洁，崔艳英，等.盐酸替利定的依赖性评价[J].中国药物依赖性杂志，2006，15(5) : 363 - 366.
[10] Zhang ZZ，Gong YY，Shi YH， et al.Valproate promotes survival ofretinal ganglion cells in a rat model of optic nerve crush[J].Neuroscience，2012，224(8) : 282 - 293.
[11] Soni CR， Johnson LN.Visual neuropraxia and progressive vision loss from thyroid-associated stretch optic neuropathy[J].Eur J Ophthalmol，2010，20(2) : 429 - 436.
[12] Ma K，Xu L，Zhan H， et al.Dosage dependence of the effect of Ginkgo biloba on the rat retinal ganglion cell survival after optic nerve crush[J].Eye(Lond),2009，23(7) : 1598 - 1604.
[13] Schmued LC，Fallon JH.Fluoro-Gold: a new fluorescent retrograde axonal tracer with numerous unique properties[J].Brain Res，1986，377(1) : 147 - 154.
[14] Chiu K，Lau WM，Yeung SC， et al.Retrograde labeling of retinal ganglion cells by pplication of fluoro-gold on the surface of superior colliculus[J].J Vis Exp，2008，3791(16) : 819 - 820.

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荧光金逆行标记评价视神经切断后视网膜节细胞存活率

作者简介: 王能(1984- ),男，硕士研究生.

1. 1. 蚌埠医学院第一附属医院中心实验室,
2.
3. 2. 组织移植安徽省重点实验室, 安徽蚌埠 233004,
4.
5. 3. 山东省蓬莱市中医院眼耳鼻喉科, 265600

收稿日期: 2013-01-14
网络出版日期: 2014-02-15

基金项目: 安徽省教育厅自然科学研究重点项目(KJ2010A241)

关键词:

摘要: 目的：利用荧光金逆行标记评价视神经切断后视网膜节细胞(RGCs)的存活率。方法：将SD大鼠20只随机均分为4组,正常对照组和视神经切断后5 d、7 d和14 d组。用荧光金逆行标记和定量解剖学技术观察正常对照组和视神经切断后5 d、7 d和14 d组RGCs的密度。结果:正常对照组RGCs呈放射状分布,边界清晰,可见明显细胞突起,无渗漏现象;视神经切断组RGCs随时间延长而减少,细胞分布不均匀,并且可见吞噬死亡细胞碎片和从死亡细胞渗漏的荧光金的小胶质细胞。正常对照组左眼与右眼RGCs密度差异无统计学意义(P0.05);视神经切断后RGCs进行性减少,切断后5 d、7 d和14 d组左眼RGCs密度均明显低于正常对照组(P0.01),视神经切断后5 d、7 d和14 d存活率分别为53%、38%和13%。结论：荧光金逆行标记是评价视神经切断后RGCs存活率有效的方法。

English Abstract

Evaluating the survival rate of axotomized retinal ganglion cells by fluorogold retrograde labeling

1. 1. Department of Central Laboratory, The First Affiliated Hospital of Bengbu Medical College, Bengbu Anhui 233004;
2. 2. Department of Key Laboratory of Tissue Transplantation, Bengbu Anhui 233004;
3. 3. Department of Otolaryngology, Penglai Traditional Chinese Medicine Hospital, Penglai Shandong 265600, China

Received Date: 2013-01-14
Available Online: 2014-02-15

Abstract: Objective: To evaluate the survival rate of retinal ganglion cells(RGCs) after transsection of the optic nerve using the method of fluorogold retrograde labeling. Methods: Twenty SD rats were randomly divided inObjective: To four groups: normal control group，5 d，7 d and 14 d after transsection of the optic nerve groups. The RGCs density of the normal control group and 5 d，7 d and 14 d after transsection of the optic nerve groups was observed by fluorogold retrograde labeling and quantitative anaObjective: To mical technology. Results: In the normal control group， the RGCs labeled by the fluorescent gold presented radial distribution，clear boundary and obvious cell processes without any leak. In the optic nerve transsection groups， the number of RGCs decreased with time increasing and the cell distribution was not uniform; and the debris of dead cells and cell leakage from the dead FG microglial cells were observed. In thenormal control group， the average density of RGCs of the left eye had no significant difference compared Objective: To that of the right eye(P ＞0. 05). In the 5 d，7 d and 14 d after transsection of the optic nerve groups， the number of RGCs was reduced progressively and the density of RGCs of the left eye was significantly lower than that in the normal control group(P ＜ 0. 01) ; the survival rates of RGCs in the groups of 5 d，7 d and 14 d after optic nerve transsection were 53%，38% and 13%，respectively. Conclusions: luorogold retrograde labeling is an effective method of evaluating the survival rate of axoObjective: Tomized retinal ganglion cells.