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甲状腺激素诱导的神经干细胞移植对慢性实验性变态反应性脑脊髓炎的神经保护作用
杜杰 胡光强 邓莉 杨朝鲜 郭侃 高小青*
解剖学报 ›› 2015, Vol. 46 ›› Issue (1) : 13-19.
甲状腺激素诱导的神经干细胞移植对慢性实验性变态反应性脑脊髓炎的神经保护作用
Neuroprotective effect of grafting thyroid hormone-induced neural stem cells on chronic experimental allergic encephalomyelitis
目的 探讨甲状腺激素诱导的神经干细胞(NSCs)是否比单纯NSCs移植对慢性实验性变态反应性脑脊髓炎(EAE)有更好的神经保护作用。方法 体外培养新生大鼠NSCs和三碘甲腺原氨酸(T3)诱导的NSCs(T3/NSCs)，诱导其分化7d后，免疫细胞化学或免疫荧光染色分别检测半乳糖脑苷脂阳性（GalC+）和GFAP阳性（GFAP+）细胞。用豚鼠脊髓匀浆诱导慢性EAE大鼠模型。在免疫后10d，脑立体定位仪分别移植T3/NSCs、NSCs和生理盐水入EAE大鼠侧脑室，T3/NSCs和NSCs移植前用5-溴脱氧尿嘧啶核苷（BrdU)标记。实验分为T3/NSCs组、NSCs组和对照组，每组10只。每天对大鼠临床症状评分评估神经功能。大鼠免疫60d后处死，HE和LFB染色分别观察脑的炎症侵润和脱髓鞘；免疫荧光双标染色检测脑的GalC+/BrdU+和GFAP+/BrdU+比例；RT-PCR法检测脑组织血小板源性生长因子α受体（PDGFαR）、GalC和髓鞘碱性蛋白（MBP）mRNA的表达。 结果 T3/NSCs体外分化为GalC+和GFAP+细胞的比例分别高于和低于NSCs的分化。T3/NSCs组和NSCs组神经功能恢复较对照组好。T3/NSCs组较NSCs组更明显改善脑的炎症侵润和脱髓鞘，其脑内GalC+/BrdU+及GFAP+/BrdU+的比例分别高于和低于NSCs组，脑组织的PDGFαR、GalC和MBP mRNA的表达也较NSCs组高。 结论 T3/NSCs比NSCs对EAE有更好的神经保护作用。
Objective To explore whether transplantation of neural stem cells induced by thyroid hormone (T3) provides a better neuroprotective effect than native NSCs after chronic experimental allergic encephalomyelitis. Methods Neural stem cells (NSCs) and T3-induced NSCs of neonatal rat were cultured in vitro. After 7 days they were induced to differentiate, immunohistochemistry or immunofluorescence staining were used to detect GalC+ and GFAP+ cells respectively. Chronic EAE rat models were induced by Guinea pig spinal cord homogenate, fo11owed by infusion of T3/NSCs, NSCs (T3/NSCs and NSCs were labeled with 5-bromo-2’-deoxyuridine (BrdU) and saline (T3/NSCs, NSCs and control groups，respectively) at 10 days after EAE immunization, and each group had 10 rats. Clinical scores for every day were performed to evaluate neuro1ogica1 function. All rats were sacrificed at 60d after EAE immunization. HE and LFB staining was used to observe inflammatory infiltrates and demyelination in brain respectively. Immunofluorescent double staining was used to test the ratio of GalC+/BrdU+ and GFAP+/BrdU+in brain. RT-PCR assay was used to identify the expression of mRNA for PDGFαR, GalC and MBP of brain tissue. Results The ratio of GalC+ or GFAP+ cells of T3/NSCs was respectively higher or lower than that of NSCs in vitro. Significant recovery of neuro1ogica1 function was found in T3/NSCs and NSCs groups compared with control group. Improvement of inflammatory infiltrates and demyelination in T3/NSCs groups was stronger than that in NSCs groups. In brain, the ratio of GalC+/BrdU+ or GFAP+/BrdU+ in T3/NSCs group was respectively higher or lower than that in NSCs group. And the expression of mRNA for PDGFαR, GalC and MBP of brain tissue in T3/NSCs groups was stronger than in NSCs groups. Conclusion The results demonstrate that grafting T3-induced NSCs provides better neuroprotection for EAE than grafting NSCs alone.
甲状腺激素 / 神经干细胞 / 细胞移植 / 变态反应 / 脑脊髓炎 / 免疫荧光 / 大鼠
Thyroid hormone / Neural stem cell / Cell transplantation / Allergy / Encephalomyelitis / Immunofluorescence / Rat
［1］Fernandez M, Giuliani A, Pirondi S, et al. Thyroid hormone administration enhances remyelination in chronic demyelinating inflammatory disease［J］. PNAS, 2004, 101(46): 16363-16368.［2］Einstein O, Grigoriadis N, Mizrachi-Kol R, et al. Transplanted neural precursor cells reduce brain inflammation to attenuate chronic experimental autoimmune encephalomyelitis［J］. Exp Neurol, 2006, 198(2): 275-284.
［3］Fazeli AS, Nasrabadi D, Pouya A, et al. Proteome analysis of post-transplantation recovery mechanisms of an EAE model of multiple sclerosis treated with embryonic stem cell-derived neural precursors［J］. J Proteomics. 2013, 94: 437-450.
［4］Hackett C, Knight J, Mao-Draayer Y. Transplantation of Fas-deficient or wild-type neural stem/progenitor cells (NPCs) is equally efficient in treatingexperimental autoimmune encephalomyelitis (EAE)［J］. Am J Transl Res, 2014, 6(2): 119-128.
［5］Kassis I, Grigoriadis N, Gowda-Kurkalli B, et al. Neuroprotection and immunomodulation with mesenchymal stem cells in chronic experimental autoimmune encephalomyelitis［J］. Arch Neurol, 2008, 65(6): 753-761.
［6］Kassis I, Petrou P, Halimi M, et al. Mesenchymal stem cells (MSC) derived from mice with experimental autoimmune encephalomyelitis (EAE) suppress EAE and have similar biological properties with MSC from healthy donors［J］. Immunol Lett, 2013, 154(1-2): 70-76.
［7］Liu R, Zhang Z, Lu Z,et al. Human umbilical cord stem cells ameliorate experimental autoimmune encephalomyelitis by regulating immunoinflammation and remyelination［J］. Stem Cells Dev, 2013, 22(7): 1053-1062.
［8］Ghasemi N, Razavi S, Mardani M, et al. Transplantation of human adipose-derived stem cells enhances remyelination in lysolecithin-induced focal demyelination of rat spinal cord［J］. Mol Biotechnol, 2014, 56(5): 470-478.
［9］Fernández M, Paradisi M, Del Vecchio G, et al. Thyroid hormone induces glial lineage of primary neurospheres derived from non-pathological and pathological rat brain: implications for remyelination-enhancing therapies［J］. Int J Dev Neurosci, 2009, 27(8): 769-778.
［10］Kono DH, Urban JL，Horvath SJ, et al. Two minor determinants of myelin basic protein induce experimental allergic encephalomyelitis in SJL/J mice［J］. J Exp Med, 1988, 168(1): 213-227.
［11］Moore S, Khalaj AJ, Yoon J, et al. Therapeutic laquinimod treatment decreases inflammation, initiates axon remyelination, and improves motor deficit in a mouse model of multiple sclerosis［J］. Brain Behav, 2013, 3(6): 664-682.
［12］Calza L, Fernandez M, Giuliani A, et al. Thyroid hormone activates oligodendrocyte precursors and increases a myelin-forming protein and NGF content in the spinal cord during experimental allergic encephalomyelitis［J］. PNAS, 2002, 99(5): 3258-3263.
［13］Dell'Acqua ML, Lorenzini L, D'Intino G, et al. Functional and molecular evidence of myelin- and neuroprotection by thyroid hormone administration in experimental allergic encephalomyelitis［J］. Neuropathol Appl Neurobiol, 2012, 38(5): 454-470.
［14］Dugas JC, Ibrahim A, Barres BA. The T3-induced gene KLF9 regulates oligodendrocyte differentiation and myelin regeneration［J］. Mol Cell Neurosci, 2012, 50(1): 45-57.
［15］Pluchino S, Quattrini A, Brambilla E, et al. Injection of adult neurospheres induces recovery in a chronic model of multiple sclerosis［J］. Nature, 2003, 422 (6933): 688-694.
［16］Nessler J, Bénardais K, Gudi V, et al. Effects of murine and human bone marrow-derived mesenchymal stem cells on cuprizone induced demyelination［J］. PLoS One, 2013, 8(7): e69795.
［17］Proescholdt MA, Jacobson S, Tresser N, et al. Vascular endothelial growth factor is expressed in multiple sclerosis plaques and can induce inflammatory lesions in experimental allergic encephalomyelitis rats［J］. J Neuropathol Exp Neurol, 2002, 61(10): 914-925.
［18］Zappia E, Casazza S, Pedemonte E, et al. Mesenchymal stem cells ameliorate experimental autoimmune encephalomyelitis inducing T-cell anergy［J］. Blood, 2005, 106(5): 1755-1761.
［19］Liu B, Li LY, Liu ChR, et al. Effects of T3 on differentiation of human neural stem cells to oligodendrocyte［J］. Acta Anatomica Sinica, 2003, 34(2): 213-216.
刘奔, 李兰英, 刘春蓉, 等. T3对人神经干细胞分化为少突胶质细胞的影响［J］. 解剖学报，2003, 34(2): 213-216. (in Chinese)
［20］Li R, Xu W, Chen Y, et al. Raloxifene suppresses experimental autoimmune encephalomyelitis and NF-kb-dependent CCL20 expression in reactive astrocytes［J］. Plos One, 2014, 9 (4): 1-10.
四川省教育厅科研基金资助项目;四川省卫生厅科研基金资助项目;泸州医学院院级科研基金资助项目
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