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目的:探讨法舒地尔(Fasudil)对人神经母细胞瘤细胞(SH-SY5Y)氧糖剥夺(OGD)后突触损伤的影响。方法:培养SH-SY5Y细胞,分为空白对照组、OGD组、OGD+法舒地尔组,各3皿。相差光学显微镜下观察细胞突触损伤及修复的细胞形态;Western-Blot检测ROCKⅡ、磷酸化肌球蛋白磷酸酶(p-MYPT1)、突触后致密物-95(PSD-95)、突触素(Synaptophysin)等蛋白的表达情况。结果:形态学显示法舒地尔可有效修复OGD诱导的神经突触损伤。OGD组ROCKⅡ及p-MYPT1的表达明显高于空白对照组(P<0.05),突触素及PSD-95的表达明显低于空白对照组(P<0.01)。OGD+法舒地尔组ROCKⅡ及p-MYPT1表达水平低于OGD组(P<0.05),而与对照组差异无统计学意义(P>0.05);突触素和PSD-95表达水平高于OGD组(P<0.01);PSD-95表达水平高于空白对照组(P<0.05),突触素的表达与空白对照组差异无统计学意义(P>0.05),结论:法舒地尔可有效修复OGD诱导的神经突触损伤,增加PSD-95、突触素的表达,抑制ROCKⅡ及p-MYPT1的表达。
Objective: To investigate the effect of Fasudil on synaptic damage after OGD in human neuroblastoma cells (SH-SY5Y). Methods: SH-SY5Y cells were cultured and divided into blank control group, OGD group, OGD + fasudil group, and 3 each dish. The synaptic damage and cell morphology were observed under phase contrast optical microscopy. The expressions of ROCKⅡ, p-MYPT1, PSD-95 and synaptophysin (Synaptophysin) and other protein expression. RESULTS: Morphology showed Fasudil was effective in repairing OGD-induced neuronal synaptic damage. The expression of ROCKⅡ and p-MYPT1 in OGD group was significantly higher than that in blank control group (P <0.05), and the expression of synaptophysin and PSD-95 was significantly lower than that in blank control group (P <0.01). The expression of ROCKⅡ and p-MYPT1 in OGD + Fasudil group was lower than that in OGD group (P <0.05), but not in OGD + Fasudil group (P> 0.05). The expression of synaptophysin and PSD-95 was higher than OGD (P <0.01). The expression of PSD-95 was higher than that of the blank control group (P <0.05), and there was no significant difference of synaptophysin expression between the blank control group and the blank control group (P> 0.05) Effectively repair OGD-induced neuronal synaptic damage, increase the expression of PSD-95, synaptophysin, and inhibit the expression of ROCKⅡ and p-MYPT1.