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

doi:10.3969/j.issn.2095-1396.2016.04.002

Abstract

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

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.

References

[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.