骨髓间充质干细胞及补脑Ⅰ号处理后血清对小鼠海马神经元缺氧缺糖模型ICAM-1、NF200表达的影响

Effects of BMSCs and Bunao No.1 medicated serum on expression of ICAM-1 and NF200 in mouse hippocampal neurons with oxygen-glucose deprivation

目的观察骨髓间充质干细胞(BMSCs)、补脑Ⅰ号血清对小鼠海马神经元缺氧缺糖(OGD)模型神经中丝200(NF200)、细胞间黏附因子-1(ICAM-1)的影响。方法分离培养BMSCs、海马神经元,对海马神经元进行OGD造模。制备补脑Ⅰ号血清与正常血清。经MTT法筛选补脑Ⅰ号血清对海马神经元OGD模型干预的最佳浓度为10%,时间为72 h。实验分为正常组、模型组、BMSCs组、正常血清组、中药血清组、BMSCs+正常血清组、BMSCs+中药血清组。qRT-PCR检测细胞NF200、ICAM-1 mRNA表达;Western Blot检测细胞NF200、ICAM-1蛋白表达。结果与正常组比较,模型组、BMSCs组、正常血清组、BMSCs+正常血清组、中药血清组、BMSCs+中药血清组ICAM-1、NF200 mRNA及蛋白表达升高(P<0.05);与模型组比较,BMSCs组、BMSCs+正常血清组、中药血清组、BMSCs+中药血清组NF200 mRNA及蛋白表达升高,ICAM-1 mRNA及蛋白表达降低,差异有统计学意义(P<0.05);BMSCs+中药血清组较BMSCs组、正常血清组、中药血清组及BMSCs+正常血清组的NF200 mRNA及蛋白表达升高,ICAM-1 mRNA及蛋白表达减少,差异有统计学意义(P<0.05)。结论BMSCs和补脑Ⅰ号血清可能通过抑制炎症、促进神经再生及双重作用修复OGD损伤的海马神经元,两者联合应用效果优于单独使用。

Objective To observe the effects of Bunao No.1 medicated serum and bone marrow mesenchymal stem cells(BMSCs)on neurofilament 200(NF200)and intercellular adhesion factor-1(ICAM-1)expression in hippocampal neurons with oxygen-glucose deprivation(OGD).Methods BMSCs and hippocampal neurons were isolated and cultured,and hippocampal neurons were induced by OGD.Bunao No.1 medicated serum and normal serum were prepared.MTT assay showed that the optimal concentration of Bunao No.1 medicated serum for the intervention of hippocampal neurons with OGD was 10%,and the best duration was 72 hours.The cells was divided into a normal group,model group,BMSCs group,normal serum group,Bunao No.1 medicated serum group,BMSCs+normal serum group,and BMSCs+Bunao No.1 medicated serum group.The mRNA and protein expression of NF200 and ICAM-1 were detected by qRT-PCR and Western Blot,respectively.Results Compared with the normal group,the mRNA and protein expression of ICAM-1 and NF200 in the model group,BMSCs group,normal serum group,BMSCs+normal serum group,Bunao No.1 medicated serum group,and BMSCs+Bunao No.1 medicated serum group increased significantly(P<0.05).Compared with the model group,the mRNA and protein expression of NF200 significantly increased and those of ICAM-1 significantly decreased in the BMSCs group,BMSCs+normal serum group,Bunao No.1 medicated serum group,and BMSCs+Bunao No.1 medicated serum group(P<0.05).Compared with the BMSCs group,normal serum group,Bunao No.1 medicated serum group,and BMSCs+normal serum group,the mRNA and protein expression of NF200 significantly increased and those of ICAM-1 significantly decreased in the BMSCs+Bunao No.1 medicated serum group(P<0.05).Conclusion BMSCs and Bunao No.1 medicated serum can inhibit inflammation,promote nerve regeneration,and repair the hippocampal neurons damaged by OGD.Combined use of BMSCs and Bunao No.1 medicated serum is better than their single use.