乌头碱对新生大鼠心肌细胞DNA损伤的影响

目的探讨乌头碱对新生大鼠心肌细胞DNA损伤的影响。方法新生SD大鼠20只,随机分为4组,取心室肌以差速贴壁法原代培养心室肌细胞。培养第6天,制成密度为2×105个细胞/mL的细胞悬液,分别加入乌头碱混合液至终浓度为0.1、0.5、1、2μmol/L,染毒30m in;采用彗星电泳技术及CASP分析软件,检测不同浓度乌头碱染毒后心室肌细胞DNA损伤程度。结果心肌细胞被不同浓度乌头碱染毒后,尾部DNA含量、彗尾长度、尾矩、O live尾矩均随乌头碱浓度增加而升高,头部DNA含量则逐渐降低,与对照组相比,有显著性差异(P<0.01);乌头碱染毒剂量越大,心肌细胞DNA损伤越严重。结论乌头碱染毒引起细胞DNA断裂损伤呈明显的剂量—效应关系,推测细胞DNA损伤是乌头碱毒作用机制之一。

Effects of aconitine on DNA damage in myocytes of neonatal rats

】Objective To investigate the effects of aconitine on DNA damage of cultured myocytes of neonatal rats.Methods Myocytes of neonatal rats were incubated by differential adherent culture.An amount of 0.1% trypsinase PBS solution added,so that unattached cells were adjusted to a density of 2×105 cell/mL.At the 6th day,adding aconitine mixture to unattached cells at concentrations of 0.1μmol/L,0.5μmol/L,1μmol/L and 2μmol/L,were incubated for 30 min,while PBS cell suspension as normal.DNA damage of cultured myocytes of neonatal rats after aconitine incubation at different concentrations was determined by comet assay technology and CASP analysis software.Results After different doses of aconitine incubation,DNA contents,tail length,tail moment and Olive tail length of neonatal rats' myocytes all raise along with increase of aconitine dose,meanwhile head DNA contents reduce gradually;there were significant difference compared with the control group.The higher dose of aconitine incubated,the severer myocytes DNA damaged.Conclusion DNA rupture and damage induced by aconitine incubation appears to be a dose-effect relation.DNA damage might be one of aconitine toxicology mechanisms.