氯胺酮、异丙酚对谷氨酸诱导大鼠海马星形胶质细胞损伤的影响

目的:观察氯胺酮和异丙酚对谷氨酸诱导体外培养大鼠海马星形胶质细胞损伤的影响。方法:取出生1~3 d Wistar大鼠海马星形胶质细胞,原代纯化培养3周。将细胞随机分为5组(n=9):C组为对照组,加入Hanks液;G组加入谷氨酸;GK组先加入谷氨酸,10 min后加入氯胺酮;GP组先加入谷氨酸,10 min后加入异丙酚;GPK组先加入谷氨酸,10 min后同时加入异丙酚和氯胺酮。培养24 h后分别检测各组上清液IL-1β、TNF-α和IL-10浓度、细胞凋亡率及胞内SOD活性和MDA含量,并观察细胞形态学改变。结果:与C组比较,G组星形胶质细胞凋亡增加(P<0.01),未凋亡的细胞被激活增生肥大,IL-1β和TNF-α浓度升高(P<0.01),IL-10浓度无明显变化(P>0.05),SOD活性显著降低(P<0.01),MDA含量明显增加(P<0.01)。与G组比较,GK、GP和GPK组凋亡细胞减少(P<0.05,P<0.01),IL-1β和TNF-α降低(P<0.05,P<0.01),IL-10升高(P<0.01),SOD活性增加而MDA含量低(P<0.05,P<0.01),细胞无明显增生肥大。GK组和GP组间比较差异无统计学意义(P>0.05),GP组和GK组分别与GPK组比较差异有统计学意义(均P<0.01)。结论:氯胺酮和异丙酚均可抑制谷氨酸引起的大鼠海马星形胶质细胞的凋亡和激活,通过抑制脂质过氧化反应,清除自由基,同时抑制炎性细胞因子分泌而发挥神经保护作用,且两者有协同作用。

Effects of Propofol and Ketamine on Glutamate-induced Injury of primary Cultured Hippocampal Astrocytes of Rats

Objective To investigate the effects of propofol and ketamine on the injury in primary cultured hippocampal astrocytes of rats induced by glutamate.Methods Astrocytes were taken from hippocampus of 2~3 days old Wistar rat and cultured.The cells were treated with the following five conditions(n=9): ①group-C: control group: sham wash with Hanks solution;②group-G: 100μmol/L of glutamate was added;③ group-GK: 100μmol/L of Glutamate was first added,and 10 minutes later 50μmol/L of ketamine was then added;④ group-GP: 100 μmol/L of glutamate was first added,and 10 minutes later 50 μmol/L of propofol was then added;⑤ group-GPK: 100 μmol/L of glutamate was first added,and 10 minutes later added propofol 50μmol/L and ketamine 50μmol/L simultaneously.After 24 hours of incubation,the concentrations of IL-1β,TNF-α and IL-10 in the supernatant,the contents of malondialdehyde(MDA) and activity of intracellular superoxide dismutase(SOD) were measured and the morphological changes were observed with wright staining,the apoptotic cell death was analyzed with flow cytometer.Results Apoptotic cells death was significantly increased,the concentrations of IL-1 β and TNF-α were also significantly increased,the contents of MDA was significantly increased and the activity of SOD was markedly decreased in Group-G compared with Group-C(P<0.01),the IL-10 remained low in Group G(P>0.05).The apoptotic astrocytes was lower,IL-1 β and TNF-α were also significantly lower and the contents of MDA were significantly lower and the activity of SOD significantly increased in Group-GK(P<0.05) and Group-GP(P<0.05) and Group-GPK(P<0.01) than in Group-G,but the IL-10 was greatly higher in the two groups compared with that in Group G(P<0.01),the cells were eumorphism.There is obviously difference in Group-GPK compared with Group-GK or Group-GP respectively(P<0.01) and no difference in Group-GK and Group-GP.Conclusion Propofol and ketamine inhibit the injury of hippocampal astrocytes induced by glutamate in a synergistic action.The anti-injury mechanisms may be explained by inhibiting the secretion of IL-1 β and TNF-α yet increase IL-10 synthesis and release,and clearing oxygen free radcial and increasing the activity of antioxidase and inhibiting lipid peroxidation.