促红细胞生成素与甲基强的松龙对星形胶质细胞的作用研究  

Effect of Erythropoietin and Methylprednisolone on Astrocytes

作　　者：郝彦明[1] 王洪震[1] 贾正平[1] 徐又佳[2] 俞晨[2] 

机构地区：[1]江苏省昆山市第一人民医院关节科,江苏苏州215300 [2]苏州大学附属第二医院骨科,江苏苏州215004

出　　处：《中国药业》2015年第10期23-26,共4页China Pharmaceuticals

摘　　要：目的研究甲基强的松龙(MPSS)与促红细胞生成素(EPO)联合应用对体外培养的原代星形胶质细胞的作用。方法采用出生3 d内SD新生鼠的大脑制成细胞悬浊液后,以含15%胎牛血清的DMEM培养液培养,摇床后逐渐传代、纯化星形胶质细胞,磷酸盐缓冲液(PBS)代替培养基模拟缺营养模型3 h后即刻分别加入MPSS 10μmol/L,EPO 10 U/L,MPSS 10μmol/L+EPO 10 U/L,分别继续培养3 d,采用免疫荧光技术鉴定星形胶质细胞,MTT法检测细胞的增殖活性和凋亡,聚合酶链式反应(PCR)技术测定细胞胶质纤维酸性蛋白(GFAP)m RNA表达水平。结果星形胶质细胞摇床后传至第3代,经鉴定细胞纯度达95%以上;缺营养3 h后部分细胞形态变圆甚至死亡漂浮于正常细胞表面;正常组和损伤组分别加入MPSS,EPO,MPSS+EPO 3 d后,损伤组细胞活性与GFAP m RNA表达水平均明显升高(P<0.05),且MPSS和EPO联合应用与单用MPSS、单用EPO相比,细胞活性与GFAP m RNA表达水平明显升高(P<0.05),但正常组无明显变化(P>0.05)。结论适当剂量的MPSS与EPO联合应用对缺营养损伤星形胶质细胞有显著的保护作用,且与单用MPSS、单用EPO相比有显著性差异,但对正常星形胶质细胞无显著的保护作用。Objective To study the effect of methylprednisolone（MPSS） combined with erythropoietin（EPO） to in vitro cultured primary astrocytes（AST）. Methods The cell suspension was prepared by using the cerebrum of neonatal rats within 3 d of birth,cultured by the DMEM culture solution containing 15% fetal calf serum,gradually passaged after shaking for purifying astrocytes. After simulating nulli-nutrition model for 3 h by phosphate buffer solution（PBS） for substituting medium,MPSS 10 μmol/L,EPO 10 U/L and MPSS 10 U/L plus EPO 10 U/L were immediately added and the continuous culture for 3 d was performed. Astrocytes were identified by the immunofluorescent technique;the proliferation activity and apoptosis of astrocytes were detected by MTT;the expression level of GFAP mRNA was detected by PCR. Results Astrocytes were passaged to the third generation after shaking,the astrocytes purity reached more than 95% by identification;part of astrocytes became round in morphology and even died after nulli-nutrition for 3 h;after 3 d of applying MPSS,EPO and MPSS plus EPO in the normal group and the damage group,both cytoactive and expression level of GFAP mRNA in the damage group were significantly increased （ P ﹤ 0. 05 ） ,moreover compare with the single application of MPSS or EPO,the cytoactive and GFAP mRNA expression level in the combined application of MPSS and EPO were significantly increased（ P ﹤ 0. 05）,but no obvious change was found in the normal group（ P ﹥ 0. 05）. Conclusion The combined application of proper dose of MPSS and EPO has obviously protection effect on nulli-nutrition damaged astrocytes,moreover has significant difference compared with the single appli-cation of MPSS or EPO,but has no obvious protective effect on normal astrocytes.

关 键 词：甲基强的松龙 促红细胞生成素 星形胶质细胞 缺营养 联合用药 

分 类 号：R965[医药卫生—药理学] R977[医药卫生—药学]