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目的:探讨芬戈莫德(FTY720)对实验性自身免疫性脑脊髓炎(EAE)大鼠基质金属蛋白酶(MMPs)表达的影响。方法 :按随机数表法将Wistar大鼠随机分成6组(正常对照组、EAE模型对照组、FTY720高剂量组、FTY720中剂量组、FTY720低剂量组、阳性对照组),每组10只。除正常对照组外,其余各组采用豚鼠全脊髓匀浆诱导造模,造模第15天,除正常对照组及EAE模型对照组外,其余4组分别给予高(6.17 mg/mL)、中(3.09 mg/mL)、低(1.54 mg/mL)剂量FTY720及环孢菌素(3.70 mg/mL),每天观察大鼠神态及形体变化,记录大鼠体质量及神经功能评分情况。给药2周后处死大鼠,取脑和脊髓进行HE组织染色观察病理改变,并采用免疫组化方法检测MMPs蛋白的表达。结果:(1)EAE模型对照组神经功能评分(3.50±0.71)分,FTY720各剂量给药组神经功能评分均明显低于EAE模型对照组(P<0.01),FTY720高剂量组神经功能评分(0.61±0.48)分,与阳性对照组神经功能评分(0.68±0.48)分比较差异无统计学意义(P>0.05);(2)HE组织染色:EAE模型对照组免疫后大鼠的脑组织和脊髓组织均有多个部位的血管炎,炎性细胞聚集在血管周围形成“袖套”样改变,白质改变较为显著。在FTY720各剂量给药组大鼠血管炎范围、数量及程度已明显减轻,其中FTY720高剂量组及阳性对照组的脑和脊髓形态较接近正常对照组;(3)MMPs检测发现正常对照组中MMP-2、MMP-9表达呈阴性,EAE模型对照组呈阳性,FTY720各剂量给药组中,胞浆褐色明显减少,其中FTY720高、中剂量组较接近正常对照组。结论:FTY720能够抑制EAE大鼠发病过程中MMPs蛋白的表达,起到改善EAE大鼠神经功能的作用。
Objective: To investigate the effect of fingolimod (FTY720) on the expression of matrix metalloproteinases (MMPs) in experimental autoimmune encephalomyelitis (EAE) rats. Methods: Wistar rats were randomly divided into 6 groups (normal control group, EAE model control group, FTY720 high dose group, FTY720 middle dose group, FTY720 low dose group and positive control group) according to random number table. Except for the normal control group, all the other groups were induced with whole spinal cord homogenate of guinea pigs. On the 15th day after modeling, except the normal control group and the EAE model control group, the other 4 groups were given high (6.17 mg / mL) (3.09 mg / mL) and low dose (1.54 mg / mL) of FTY720 and cyclosporin (3.70 mg / mL). The morphological and morphological changes of rats were observed daily. The body weight and neurological score were recorded. Two weeks after the administration, the rats were sacrificed and the brain and spinal cord were taken for HE staining to observe the pathological changes. The expression of MMPs protein was detected by immunohistochemistry. Results: (1) The neurological scores of EAE model group (3.50 ± 0.71) and FTY720 group were significantly lower than those of EAE model group (P <0.01), and those of FTY720 high dose group 0.61 ± 0.48), which was not significantly different from that of the positive control group (0.68 ± 0.48) (P> 0.05). (2) HE staining: EAE model group Spinal cord tissue has multiple parts of vasculitis, inflammatory cells gathered around the blood vessels to form “sleeve ” -like changes, the white matter changes more pronounced. The number and extent of vasculitis in each dose group of FTY720 were significantly reduced, and the shape of brain and spinal cord of FTY720 high-dose group and positive control group were closer to the normal control group. (3) MMPs detected in normal control group The positive expression of MMP-2 and MMP-9 in EAE model group was positive in EAE model group. The cytoplasm brown color of FTY720 group was significantly decreased, and FTY720 high and middle dose groups were closer to normal control group. Conclusion: FTY720 can inhibit the expression of MMPs in EAE rats and play an important role in improving neurological function in EAE rats.