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[摘要]
目的 建立SH-SY5Y细胞缺糖缺氧损伤模型,考察注射用血塞通对缺糖缺氧损伤SH-SY5Y细胞存活、凋亡及Lingo-1 mRNA及蛋白表达的影响。方法 采用RPMI 1640无糖培养基和三气培养箱缺氧法制备SH-SY5Y细胞缺糖缺氧损伤模型,CCK8法测定细胞存活率,确定最佳缺氧时间及最适注射用血塞通质量浓度,Annexin V-FITC/PI双染法检测SH-SY5Y细胞凋亡率,并通过实时荧光定量PCR(qRT-PCR)方法测定Lingo-1 mRNA表达情况,Western blotting法测定Lingo-1蛋白表达情况。结果 SH-SY5Y细胞缺糖缺氧损伤模型的最佳缺氧时间为16 h,注射用血塞通最适质量浓度为640 mg/L,在该质量浓度条件作用下,血塞通组与模型组相比,SH-SY5Y细胞的凋亡率显著降低,Lingo-1的mRNA表达水平及蛋白表达水平均显著降低。结论 当SH-SY5Y细胞受到缺糖缺氧损伤后,细胞凋亡率显著升高,Lingo-1呈高表达状态,血塞通可以明显减少缺糖缺氧损伤SH-SY5Y细胞的凋亡,同时抑制Lingo-1的高表达,具有抑制凋亡和显著的神经保护作用。
[Key word]
[Abstract]
Objective To establish an oxygen and glucose deprivation model (OGD) of SH-SY5Y cells, and investigate the effects of Xuesaitong Injection on the cell survival, apoptosis rate of SH-SY5Y cells and the mRNA and protein expressions of Lingo-1. Methods The model was established using 1640 sugar-free medium and three-gas incubator. The cell survival rate was determined by CCK8 method to determine the optimal time for hypoxia and the optimal concentration of Xuesaitong Injection. The apoptosis rate of SH-SY5Y cells was detected by Annexin V-FITC/PI double staining. The mRNA and protein expressions of Lingo-1 were determined by qRT-PCR and Western blotting, respectively, and the effect of Xuesaitong on Lingo-1 expression was finally determined. Results In this study, the optimal hypoxia time for the establishment of the OGD model of SH-SY5Y cells was 16 h and the optimum concentration of Xuesaitong Injection was 640 mg/L. The apoptosis rate of SH-SY5Y cells was significantly reduced, and the expression levels of lingo-1 mRNA and protein were decreased in the Xuesaitong group compared with the model group under the condition of this concentration. Conclusion The apoptosis rate was significantly increased and Lingo-1 was highly expressed when SH-SY5Y cells were damaged by oxygen and glucose deprivation. Xuesaitong can significantly reduce the apoptosis of SH-SY5Y cells induced by oxygen and glucose deprivation, and inhibit the high expression of lingo-1, which has the anti-apoptosis and significant neuroprotective effect.
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[基金项目]
国家自然科学基金面上资助项目(81573926);国家自然科学基金面上资助项目(81173235);国家自然科学基金面上资助项目(30772803);北京中医药大学在读研究生项目(2018-JYBZZ-XS144)