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目的:观察鞘氨醇激酶1(SPK1)基因转染的脐带间充质干细胞(UCMSC)移植对实验性自身免疫性脑脊髓炎(EAE)小鼠的疗效。方法:将C57BL/6小鼠分为对照组(12只)、EAE组、UCMSC组和UCMSC-SPK1组;后3组进行免疫诱导以制备EAE模型,每组11只。以第一次注射抗原当日为第0天,UCMSC组和UCMSC-SPK1组小鼠分别在免疫后第14、21、28天通过尾静脉注射UCMSC和UCMSC-SPK1,对照组和EAE组注射同等体积的生理盐水。每日采用双盲法进行小鼠神经功能缺损评分;第33天,ELISA法检测外周血IFN-γ、TNF-α、IL-17和L-4水平,分别采用HE和Fast-blue(LFB)染色观察并进行脊髓炎性细胞浸润评分、脱髓鞘评分,免疫组化法检测脊髓GFAP和MBP的表达,流式细胞术检测脾脏中调节性T细胞和NK细胞比例。结果:与EAE组比较,UCMSC-SPK1组和UCMSC组小鼠神经功能缺损评分、脊髓炎性细胞浸润评分和脱髓鞘评分降低,脾脏调节性T细胞比例升高、NK细胞比例降低;UCMSC-SPK1组变化更为显著(P<0.05);UCMSC-SPK1组脊髓GFAP表达较EAE组下降,MBP表达增强(P<0.05),外周血IFN-γ、TNF-α、IL-17水平降低,IL-4水平升高(P<0.05)。结论:UCMSC-SPK1移植能较UCMSC更为显著地修复EAE小鼠神经功能损伤。
OBJECTIVE: To observe the efficacy of transplanted cord blood mesenchymal stem cells (UCMSCs) transfected with sphingosine kinase 1 (SPK1) gene in experimental autoimmune encephalomyelitis (EAE) mice. Methods: C57BL / 6 mice were divided into control group (12 rats), EAE group, UCMSC group and UCMSC-SPK1 group. The latter three groups were induced by immunization to prepare EAE model. On day 0 of the first injection of antigen, UCMSC group and UCMSC-SPK1 group mice were injected with UCMSC and UCMSC-SPK1 through caudal vein on the 14th, 21st and 28th day after immunization respectively. The control group and EAE group were injected with the same volume Saline. The neurological deficit scores of mice were double-blinded daily. On the 33rd day, the levels of IFN-γ, TNF-α, IL-17 and L-4 in peripheral blood were measured by ELISA. HE and Fast- Spinal cord inflammatory cell infiltration score, demyelination score, immunohistochemistry were used to detect the expression of GFAP and MBP in spinal cord. The proportion of regulatory T cells and NK cells in spleen was detected by flow cytometry. Results: Compared with EAE group, the neurological deficit score, inflammatory cell infiltration score and demyelination score of UCMSC-SPK1 group and UCMSC group decreased, while the proportion of spleen T regulatory cells and NK cells decreased. UCMSC- (P <0.05). The expression of GFAP in UCMSC-SPK1 group was lower than that in EAE group (P <0.05), while the level of IFN-γ, TNF-α and IL-17 in peripheral blood was decreased -4 levels (P <0.05). CONCLUSIONS: UCMSC-SPK1 transplantation can repair neuronal damage more significantly than UCMSC in EAE mice.