右美托咪定对布比卡因诱发神经细胞PARP-1依赖的程序性死亡的影响

Effect of dexmedetomidine on bupivacaine-induced parthanatos in neurons

目的评价右美托咪定对布比卡因诱发神经细胞多聚腺苷二磷酸核糖聚合酶1(PARP-1)依赖的程序性死亡的影响。方法离体培养SH-SY5Y细胞株,每孔1×10^6个密度接种于6孔板,取对数期细胞采用随机数字表法分为5组(n=6):对照组(C组)、布比卡因组(B组)、不同浓度右美托咪定组(D1组、D10组、D100组)。C组正常培养;B组以布比卡因终浓度5 mmol/L孵育30 min;D1组、D10组和D100组分别以右美托咪定终浓度1、10和100 ng/ml孵育24 h,之后再用布比卡因终浓度5 mmol/L孵育30 min。各组培养或孵育结束时,采用MTT法检测细胞存活率,采用Western blot法检测PARP-1和多聚腺苷二磷酸核糖(PAR)表达水平。结果与C组比较,B组、D1组、D10组和D100组细胞存活率降低,PARP-1以及PAR表达上调(P<0.05);与B组比较,D1组、D10组和D100组细胞存活率升高,PAR及PARP-1表达下调(P<0.05);与D1组比较,D10和D100组细胞存活率升高,PAR及PARP-1表达下调(P<0.05)。结论右美托咪定可抑制布比卡因诱发的PARP-1依赖的程序性死亡,减少神经细胞损伤。

Objective To evaluate the effect of dexmedetomidine on bupivacaine-induced poly(ADP-ribose)(PAR)polymerase-1(PARP-1)-dependent programmed death(parthanatos)in neurons.Methods SH-SY5Y cell lines were cultured in vitro and inoculated on 6-well plates at a density of 1×10^6 cells per well.Cells at the logarithmic growth phase were divided into 5 groups(n=6 each)by a random number table method:control group(group C),bupivacaine group(group B),and different concentrations of dexmedetomidine groups(group D1,group D10,group D100).Cells were cultured in normal culture atmosphere in group C.Cells were incubated with bupivacaine at a final concentration of 5 mmol/L for 30 min in group B.Cells was incubated with dexmedetomidine with final concentration gradients of 1 ng/ml(group D1),10 ng/ml(group D10)and 100 ng/ml(group D100)for 24 h and then with bupivacaine at a final concentration of 5 mmol/L for 30 min.At the end of culture or incubation,the cell survival was measured by MTT assay,and the expression of PARP-1 and PAR was determined by Western blot.The cell survival rate was calculated.Results Compared with group C,the cell survival rate was significantly decreased,and the expression of PARP-1 and PAR was up-regulated in B,D1,D10 and D100 groups(P<0.05).Compared with group B,the cell survival rate was significantly increased,and the expression of PARP-1 and PAR was down-regulated in D1,D10 and D100 groups(P<0.05).Compared with group D1,the cell survival rate was significantly increased,and the expression of PARP-1 and PAR was down-regulated in D10 and D100 groups(P<0.05).Conclusion Dexmedetomidine can inhibit PARP-1-dependent programmed death induced by bupivacaine and reduce damage to neurons.