氯胺酮对谷氨酸诱导大鼠脊髓背角星形胶质细胞内游离钙浓度的影响

目的:观察氯胺酮对谷氨酸诱导大鼠脊髓背角星形胶质细胞凋亡和胞内游离钙浓度([Ca2+]i)变化的影响。方法:取新生2~3 d W istar大鼠T11~L6脊髓背角星形胶质细胞,原代纯化培养。将细胞随机分为:对照组(C组),谷氨酸组(G组),氯胺酮组(K组),余下3组为谷氨酸+不同浓度氯胺酮组(标记为GK1~GK3组),培养30 m in后取各细胞检测[Ca2+]i,再培养48 h检测星形胶质细胞凋亡率。结果:与C组比较,G组细胞发生了大量凋亡(P<0.01),[Ca2+]i显著升高(P<0.01);与G组比较,GK2组细胞凋亡率和[Ca2+]i明显降低(P<0.05),GK3组细胞凋亡率和[Ca2+]i显著降低(P<0.01)。结论:适量氯胺酮通过抑制细胞内钙超载显著抑制了谷氨酸诱导星形胶质细胞凋亡。

The Effect of Ketamine on Glutamate-induced [Ca2+]I in astrocyte cells in Rat Spinal Cord Dorsal Horn

Objective To observe the effect of ketamine on Glutamate-induced apoptosis and the intracellular dissociated Ca~(2+) concentration([Ca~(2+)]i) in rat cultured spinal cord dorsal horn astrocytes.Methods Cells were derived from T_(11)-L_6 spinal cord dorsal horn of 2-3 days old Wistar rat and purified for two weeks,they were treated with following six conditions:(1)sham wash with Hanks solution(control group-C);(2)100 μmol/L of Glutamate(group-G);(3)100 μmol/L ketamine(group-K);(4-6) 100 μmol/L of Glutamate was added and 30 mins later,ketamine at 10,50 or 100 μmol/L was superimposed,respectively(group GK_1-GK_3).After 30 mins or 48 hrs treatment,the [Ca~(2+)]i and the apoptotic cell death was analyzed with flow cytometer.Results Apoptotic cell death and the [Ca~(2+)]i was significantly increased in Group G when compared with Group C(P<0.01).The apoptotic astrocytes and the [Ca~(2+)]i was significant low in GK_2 group(P<0.05) and GK_3 group(P<0.01) than in Group G.There was no effect of ketamine itself at 100 μmol/L on apoptotic cell death and [Ca~(2+)]i.Conclusion Ketamine at 100 μmol/L can inhibit glutamate-induced apoptosis cell death in rat cultured spinal cord dorsal horn astrocytes.Its anti-apoptotic mechanisms may be explained by inhibiting the intracellular Ca~(2+) overloading.