摘要
目的:观察不同浓度尼卡地平对谷氨酸诱导大鼠海马星形胶质细胞损伤的影响。方法:取新生2～3 dSD大鼠海马星形胶质细胞,原代纯化培养3周。将细胞随机分6组(n=9):正常对照组(C组)加入Hanks液;谷氨酸组(G组)加入谷氨酸至终浓度500μmol/L;尼卡地平组(N组)加入尼卡地平至终浓度10μmol/L;GN1、GN2、GN3组先加入谷氨酸至终浓度500μmol/L,10 m in后分别加入尼卡地平至终浓度1、5、10μmol/L。培养30 m in后检测海马星形胶质细胞内游离钙浓度([Ca2+]i),继续培养24 h检测各组细胞凋亡及丙二醛(MDA)含量、超氧化物歧化酶(SOD)和谷胱甘肽(GSH)活性,免疫荧光细胞化学法检测胶质纤维酸性蛋白(GFAP)表达并观察细胞形态学的改变。结果:与C组比较,G组和GN1组细胞大量凋亡,未凋亡的星形胶质细胞增生肥大;GFAP表达、[Ca2+]i及MDA含量升高,SOD和GSH活性均降低(P<0.01);G、GN1组间比较差异无统计学意义(P>0.05)。与G组比较,GN2组和GN3组凋亡明显减少,细胞形态正常;GFAP表达、[Ca2+]i及MDA含量降低,SOD和GSH活性升高。结论:尼卡地平通过抑制细胞内钙超载和脂质过氧化反应,清除自由基而抑制了谷氨酸诱导海马星形胶质细胞损伤,其抑制程度与剂量有关。
Objective To investigate the effects of different concentrations nicardipine on glutamate induced-injury of primary cultured rats hippocampal astrocytes.Methods Astrocytes were taken from hippocampus of 2～3 days old SD rats and incubated for 3 weeks.The cells were randomly divided into six groups(n=9):normal control group were given Hanks solution(group C),glutamate group(group G) were given glutamate till the final concentration to 500 μmol/L,nicardipine group(group N) were given nicardipine till the final concentration to 10 μmol/L,glutamate combined nicardipine groups(group GN1,GN2,GN3) were given glutamate till the final concentration to 500 μmol/L and then added nicardipine till the final concentration were 1,5,10 μmol/L 10 minutes later.After cultured for 30 minutes,the intracellular dissociated Ca2+ concentration(i) were determined,and then cultured for 24 hours,the cells apoptosis was analyzed with flow cytometer,the contents of malondialdehyde(MDA),activity of superoxide dismutase(SOD) and glutathione(GSH) intracellular were measured.Glial fibrillary acidic protein(GFAP) expression and the morphological changes were observed with immunofluorescence staining.Results Compared with group C,the apoptotic cells in group G and group GN1 were the majority,the non-apoptotic astrocytes were hyperplasia and hypertrophy,and the GFAP expression,i,content of MDA were significantly increased,the activity of SOD and GSH were significantly decreased.Compared with group G,the cell apoptosis were significantly decreased in group GN2 and GN3,and the GFAP expression,i,content of MDA were significantly decreased,the activity of SOD and GSH were significantly increased.Conclusion Nicardipine could inhibit the glutamate induced injury of hippocampal astrocytes through decreasing the intracellular Ca2+ overload and lipid peroxidation,clearing oxygen free radcials.
出处
《湖北医药学院学报》
CAS
2011年第4期351-355,共5页
Journal of Hubei University of Medicine
