FK506 和 FK1706 在体外对免疫抑制和促进神经轴突再生作用影响的对比研究

doi:10.3969/j.issn.2095-1396.2016.04.002

神经药理学报 ›› 2016, Vol. 6 ›› Issue (4): 13-18.DOI: 10.3969/j.issn.2095-1396.2016.04.002

FK506 和 FK1706 在体外对免疫抑制和促进神经轴突再生作用影响的对比研究

郑益新，徐杰

福建省立医院骨科，福建医科大学省立临床医学院，福州，350001，中国

出版日期:2016-08-26 发布日期:2016-08-31
通讯作者: 徐杰，男，教授，主任医师，硕士生导师；E-mail：jiexud@126.com
作者简介:郑益新，男，住院医师，硕士研究生；E-mail：yixinzheng@yeah.net
基金资助:
福建省科技计划项目重点项目（No. 2014Y0052）

Effects of FK506 and FK1706 on Immunosuppression and Axonal Regeneration in Vitro: a Comparative Study

ZHENG Yi-xin，XU Jie

The Orthopedics of Fujian Provincial Hospital，Provincial College of Clinical Medicine Fujian Medical University，Fuzhou，350001，China

Online:2016-08-26 Published:2016-08-31
Contact: 徐杰，男，教授，主任医师，硕士生导师；E-mail：jiexud@126.com
About author:郑益新，男，住院医师，硕士研究生；E-mail：yixinzheng@yeah.net
Supported by:
福建省科技计划项目重点项目（No. 2014Y0052）

摘要/Abstract

摘要： 目的：通过比较不同浓度他克莫司（tacrolimus，FK506）和 FK1706 对外周血单个核细胞（peripheral blood mononuclear cell，PBMC）的免疫抑制及对人神经母细胞瘤细胞（neuroblastoma cell，SH-SY5Y）的神经轴突再生作用，比较 FK506 和 FK1706 免疫抑制和促进神经轴突再生作用的差异。方法：用不同浓度的 FK506 和FK1706 处理 PBMC，分设 9 组：空白对照组，4 组不同浓度 FK506（0.1、1.0、10.0、100.0 nmol · L -1 ），4 组不同浓度 FK1706（0.1、1.0、10.0、100.0 nmol · L -1 ），比较各组 PBMC 的体外增殖的抑制率和上清液中的白细胞介素 -2（interleukin-2，IL-2）的含量；用不同浓度的 FK506 和 FK1706 处理 SH-SY5Y，分设 9 组：空白对照组，4 组不同浓度 FK506（0.1、1.0、10.0、100.0 nmol · L -1 ），4 组不同浓度 FK1706 （0.1、1.0、10.0、100.0 nmol · L -1 ）；用倒置显微镜观察比较各组的突触长度的变化。结果：FK506 10.0 nmol · L -1 和 100.0 nmol · L -1 组干预的 PBMC 细胞全部凋亡，0.1 nmol ·L -1 和 1.0 nmol ·L -1 FK506 对 PBMC 体外增殖和 IL-2 的分泌均表现出显著的抑制作用，呈浓度依赖性和时间依赖性（P<0.05）；实验组不同浓度 FK1706 对 PBMC 体外增殖和 IL-2 的分泌均表现出一定程度的抑制作用，与空白对照组相比，0.1 nmol ·L -1 差异无统计学意义，在 1.0~100.0 nmol ·L -1 浓度范围内随 FK1706 浓度的增加、作用时间的延长，对 PBMC 体外增殖和 IL-2 分泌的抑制作用逐渐加强，与空白对照组相比差异具有统计学意义（P<0.05），但较同浓度 FK506 实验组相比，FK1706 对 PBMC 体外增殖和 IL-2 的分泌抑制作用明显减弱（P<0.05）。在10 nmol ·L -1 浓度组，FK506和FK1706均具有最佳的促SH-SY5Y细胞突起生长作用（P<0.01）。与空白对照组相比，实验组不同浓度 FK506 和 FK1706 均能有效促进 SH-SY5Y 细胞突起生长（P<0.05），相同浓度 FK506 与 FK1706 促SH-SY5Y 细胞突起生长并无差异（P>0.05）。结论：FK1706 去除免疫抑制作用而保留促进神经再生作用。

关键词: 周围神经损伤, 神经再生, FK506 , FK1706

Abstract: Objective：To test the different effects of FK506 and FK1706 on immunosu-ppression in peripheral blood mononuclear cells and nerve regeneration in SH-SY5Y，to make sureFK1706 decrease immunosuppression while promoting nerve regeneration. Methods：Different doses of FK506 and FK1706 intervention in peripheral blood mononuclear cells were divided into nine groups：control group，different dose of FK506 group （0.1，1.0，10.0，100.0 nmol ·L -1 ），different doses of FK1706 group （0.1，1.0，10.0，100.0 nmol·L -1 ） and cell proliferation were evaluated by MTT while IL-2 levels in PBMC were evaluated by ELISA. Different dose of FK506 and FK1706 intervention SH-SH5Y cells，divided into nine groups：sham group，different concentrations of FK506 group （0.1，1.0，10.0，100.0 nmol·L -1 ），different concentrations of FK1706 group （0.1，1.0，10.0，100.0 nmol·L -1 ）. The number of cells，changes in synaptic cell body diameter and the axonal length were observed by the inverted microscope. Results：FK506 at the dose of 10.0 nmol·L -1 and 100.0 nmol·L -1 induced cell death in PBMCs. The dose of 0.1 nmol·L -1 and 1.0 nmol·L -1 reduced the cell proliferation and IL-2 secretion in concentration-and time-dependent manners （P<0.05）. FK1706 induced the decrease of PBMC proliferation and IL-2 secretion. Compared to the control group，0.1 nmol·L -1 was not statistically significant，1.0 nmol·L -1 ，10.0 nmol·L -1 ，100.0 nmol·L -1 showed after the increase of concentration and the
prolongation of the action time，the inhibitory effects of FK1706 on the cell proliferation and IL-2 secretion were signifi cantly enhanced （P<0.05），but the immunosuppression compared to the same dose of FK506 was signifi cantly attenuated （P<0.05）. Compare to the sham group，both FK506 and FK1706 could effectively promote neurite outgrowth in SH-SY5Y cells （P<0.05），there was no signifi cant difference between the two drugs （P>0.05）. Conclusion：FK1706 decrease immunosuppression while promote nerve regeneration，which may be a benefi cial strategy for nerve injury.

Key words: peripheral nerve injuries, nerve regeneration, FK506, FK1706

郑益新, 徐杰. FK506 和 FK1706 在体外对免疫抑制和促进神经轴突再生作用影响的对比研究[J]. 神经药理学报, 2016, 6(4): 13-18.

ZHENG Yi-xin, XU Jie. Effects of FK506 and FK1706 on Immunosuppression and Axonal Regeneration in Vitro: a Comparative Study[J]. ACTA NEUROPHARMACOLOGICA, 2016, 6(4): 13-18.

参考文献

[1]Li R, Liu Z, Pan Y, et al. Peripheral nerve injuries treatment: a systematic review [J]. Cell Biochemistry and Biophysics, 2014, 68(3): 449-454.

[2]Olawale A R Sulaiman, Tessa Gordon. Effects of short- and long-term Schwann cell denervation on peripheral nerve regeneration, myelination, and size [J]. Glia, 2000,32(3):234-46.

[3]Gold B G, Storm-Dickerson T, Austin D R. The immunosuppressant FK506 increases functional recovery and nerve regeneration following peripheral nerve injury [J]. Restorative Neurology and Neuroscience, 1994, 6(4): 287-96.

[4]Tai P K, Albers M W, Chang H, et al. Association of a 59-kilodalton immunophilin with the glucocorticoid receptor complex [J]. Science (New York, NY), 1992, 256(5061): 1315-18.

[5]Raymond D Price, Takayuki Yamaji, Hiroko Yamamoto, et al. FK1706, a novel non-immunosuppressive immunophilin: neurotrophic activity and mechanism of action [J]. J European Pharmacology, 2005, 509(1): 11-19.

[6]Girish N Vyas. Human peripheral blood mononuclear cell substrate for propagating wild type HIV-1[J]. Dev Biol, 2001,106: 345-356.

[7] Deng Ling-xiao, Deng Ping, Ruan Yi-wen, et al. A novel growth-promoting pathway formed by GDNF-overexpressing Schwann cells promotes propriospinal axonal regeneration, synapse formation, and partial recovery of function after spinal cord injury[J]. J Neurosci, 2013. 33(13): 5655-5667.

[8] Aldape R A, Futer O, DeCenzo M T, et al. Charged surface residues of FKBP12 participate in formation of the FKBP12-FK506-calcineurin complex [J]. J Biological Chemistry, 1992, 267(23): 16029-16032.

[9] Schreiber S L, Crabtree G R. The mechanism of action of cyclosporin A and FK506 [J]. Immunology Today, 1992, 13(4): 136-142.

[10] Roberta K Yang, James B Lowe, Julia B Sobol, et al. Dose-dependent effects of FK506 on neuroregeneration in a rat model[J]. Plastic and Reconstructive Surgery, 2003, 112(7): 1832-1840.

[11]Jean-Francois Chabas, Olivier Alluin, Guillaume Rao, et al. FK506 induces changes in muscle properties and promotes metabosensitive nerve fiber regeneration[J]. J Neurotrauma, 2009, 26(1): 97-108.

[12]Julia B Sobol, James B Lowe, Robin K Yang, et al. Effects of delaying FK506 administration on neuroregeneration in a rodent model[J]. J Reconstructive Microsurgery, 2003, 19(02): 113-118.

[13]John N Jensen, Michael Brenner, Thomas H Tung, et al. Effect of FK506 on peripheral nerve regeneration through long grafts in inbred swine[J].Annals of Plastic Surgery, 2005, 54(4): 420-427.

[14]Esther Udina, Dolores Ceballos, Bruce G Gold B, et al. FK506 enhances reinnervation by regeneration and by collateral sprouting of peripheral nerve fibers[J]. Experimental Neurology, 2003, 183(1): 220-231.

[15] Ansselin A D, Pollard J D. Immunopathological factors in peripheral nerve allograft rejection: quantification of lymphocyte invasion and major histocompatibility complex expression [J]. J Neurological Sciences, 1990, 96(1): 75-88.

[16] Daniel L Riggs, Marc B Cox, Heather L Tardif, et al. Noncatalytic role of the FKBP52 peptidyl-prolyl isomerase domain in the regulation of steroid hormone signaling [J]. Molecular Cellular Biology, 2007, 27(24): 8658-8669.

[17] Andrew M Cameron, J P Steiner, D M Sabatini, et al. Immunophilin FK506 binding protein associated with inositol 1,4,5-trisphosphate receptor modulates calcium flux [J]. Proceedings of the National Academy of Sciences of the United States of America, 1995, 92(5): 1784-1788.

[18] Sanjay C Keswani, Brian Rosenberg, Ahmet Hoke. The use of GAP-43 mRNA quantification in high throughput screening of putative neuroprotective agents in dorsal root ganglion cultures [J]. J Neuroscience Methods, 2004, 136(2): 193-95.

[19] Gold B G. FK506 and the role of the immunophilin FKBP-52 in nerve regeneration [J]. Drug Metabolism Reviews, 1999, 31(3): 649-663.

[20] Tai P K, Albers M W, Chang H, et al. Association of a 59-kilodalton immunophilin with the glucocorticoid receptor complex [J]. Science (New York, NY), 1992, 256(5061): 1315-18.

[21] Wang M S, Zeleny-Pooley M, Gold B G. Comparative dose-dependence study of FK506 and cyclosporin A on the rate of axonal regeneration in the rat sciatic nerve [J]. J Pharmacology Experimental Therapeutics, 1997, 282(2): 1084-1093.

[22] Li Xiao-xia, Wang Wei, Wei Guan-xiong, et al. Immunophilin FK506 loaded in chitosan guide promotes peripheral nerve regeneration [J]. Biotechnology Letters, 2010, 32(9): 1333-1337.

[23] 柯于海, 张振伟, 冯登殿, 等.不同时段应用他克莫司纳米微球缓释颗粒促进同种异体神经移植再生的实验研究 [J]. 中华手外科杂志, 2010, 26(4): 230-233.

[24] 张桂生, 于晋辉, 邵新中, 等. FK506缓释膜应用于同种异体神经移植的实验研究 [J]. 中华手外科杂志, 2007, 23(1): 8-10.

FK506 和 FK1706 在体外对免疫抑制和促进神经轴突再生作用影响的对比研究

Effects of FK506 and FK1706 on Immunosuppression and Axonal Regeneration in Vitro: a Comparative Study

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 6

编辑推荐

Metrics

[1]	魏孟琳, 田莉, 王小琴, 邹玉安, 薛茜. 大鼠脑缺血预处理对缺血再灌注神经功能的影响[J]. 神经药理学报, 2016, 6(2): 7-13.
[2]	蒋湘云, 尚超, 李雷, 孙树峥, 李云峰, 王恒林. 四氢孕酮：抑郁和焦虑障碍治疗的潜在新靶点[J]. 神经药理学报, 2016, 6(2): 26-30.
[3]	孙炎, 宋爱霞, 薛茜, 邹玉安. 康脑液对脑缺血再灌注大鼠神经再生相关因子的影响[J]. 神经药理学报, 2015, 5(5): 7-14.
[4]	何娜，殷明，王泽剑. 成年海马神经再生与阿尔茨海默病关系的研究进展[J]. 神经药理学报, 2013, 3(6): 25-32.
[5]	金灿, 张丹参, 陈乃宏. 成年海马神经元再生与抑郁症的机制研究进展[J]. 神经药理学报, 2013, 3(2): 58-64.
[6]	戴婷, 陈中国, 段磊, 丁建花, 范益, 胡刚. Involvement of Kir6.2-composing ATP-sensitive potassium channels in phencyclidine-induced negative symptoms of schizophrenia / Kir6.2构成的ATP敏感性钾通道在苯环己哌啶诱导的精神分裂症阴性症状中的作用[J]. 神经药理学报, 2011, 1(1): 31-38.