CAJ | 학술논문

目的观察利多卡因在异氟醚对H4神经胶质瘤细胞线粒体损伤中的保护作用.方法将H4细胞分成6组:对照组、3％异氟醚(ISO)组及3％ ISO +40、60、80、100 mg/L利多卡因组.通过流式细胞仪检测细胞凋亡,电镜观察H4细胞线粒体形态,线粒体呼吸链复合酶活性的检测及线粒体膜电位检测试剂盒(JC-1)检测线粒体膜电位变化.结果 3％异氟醚使H4细胞的凋亡率由对照组的(1.6±0.1)％增加至(33.5±3.3)％(P＜0.05),分别加入40、60、80及100 mg/L利多卡因,H4细胞凋亡率分别为(24.4±4.3)％、(17.4±0.6)％、(16.0±0.4)％及(13.3±0.7)％.3％ ISO组可见线粒体出现肿胀,甚至不同程度的基质密度减少和嵴断裂以及空泡样改变;加入100 mg/L利多卡因,H4细胞线粒体形态接近对照组.复合酶Ⅳ的活性在3％异氟醚处理后明显降低(P＜0.05),加入100 mg/L利多卡因后活性明显升高(P＜0.05).3％异氟醚使H4细胞线粒体膜电位由对照组的(8.3±1.9)％降低至(2.3±0.2)％(P＜0.05),分别加入40、60、80及100 mg/L利多卡因,H4细胞凋亡率分别为(2.6±0.1)％、(3.2±0.6)％、(4.5±0.4)％及(5.6±0.3)％.100 mg/L利多卡因组H4细胞膜电位显著升高(P＜0.05).结论异氟醚可以引起线粒体损伤导致H4细胞凋亡,而足够浓度的利多卡因能够通过减轻线粒体损伤而减少H4细胞凋亡.
목적 관찰리다잡인재이불미대H4신경효질류세포선립체손상중적보호작용.방법 장H4세포분성6조:대조조、3％이불미(ISO)조급3％ ISO +40、60、80、100 mg/L리다잡인조.통과류식세포의검측세포조망,전경관찰H4세포선립체형태,선립체호흡련복합매활성적검측급선립체막전위검측시제합(JC-1)검측선립체막전위변화.결과 3％이불미사H4세포적조망솔유대조조적(1.6±0.1)％증가지(33.5±3.3)％(P＜0.05),분별가입40、60、80급100 mg/L리다잡인,H4세포조망솔분별위(24.4±4.3)％、(17.4±0.6)％、(16.0±0.4)％급(13.3±0.7)％.3％ ISO조가견선립체출현종창,심지불동정도적기질밀도감소화척단렬이급공포양개변;가입100 mg/L리다잡인,H4세포선립체형태접근대조조.복합매Ⅳ적활성재3％이불미처리후명현강저(P＜0.05),가입100 mg/L리다잡인후활성명현승고(P＜0.05).3％이불미사H4세포선립체막전위유대조조적(8.3±1.9)％강저지(2.3±0.2)％(P＜0.05),분별가입40、60、80급100 mg/L리다잡인,H4세포조망솔분별위(2.6±0.1)％、(3.2±0.6)％、(4.5±0.4)％급(5.6±0.3)％.100 mg/L리다잡인조H4세포막전위현저승고(P＜0.05).결론 이불미가이인기선립체손상도치H4세포조망,이족구농도적리다잡인능구통과감경선립체손상이감소H4세포조망.
Objective To investigate the protective effect of lidocaine (LIDO) on mitochondria damage induced by isofluranc (ISO) in H4 glioma cells.Methods The H4 glioma cells were divided into 6 groups:control group,3％ ISO group,3％ ISO +40 mg/L LIDO group,3％ ISO +60 mg/L LIDO group,3％ ISO +80 mg/L LIDO group and 3％ ISO + 100 mg/L LIDO group.The apoptosis rate was detected by the flow cytometer,mitochondria morphology was observed under electron microscope,the activity of mitochondrial respiratory chain complexes was determined,and mitochondrial membrane potential was measured by 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolcarbocyanine iodide (JC-1).Results (1) As compared with the control group,ISO significantly increased apoptosis rate of H4 glioma cells [(1.6 ±0.1)％ vs.(33.5 ±3.3)％],but the apoptosis rate decreased with the increased dose of LIDO (P ＜0.05,vs.3％ ISO group);(2) Edema of mitochondria was seen,and subsequently the rupture of crista and the reduction of quantity were found under the electron microscope in 3 ％ ISO group.The morphological changes of mitochondria in the 3％ ISO + 100 mg/L LIDO group were similar with those in the control group;(3) As compared with the control group,the activity of mitochondrial respiratory chain complexes Ⅳ was reduced significantly (P ＜ 0.05),and the activity increased after LIDO treatment (P ＜ 0.05,3％ ISO + 100 mg/L LIDO vs.3％ ISO group);(4) Mitochondrial membrane potential was markedly decreased after treatment with ISO [(8.3 ± 1.9) ％ vs.(2.3 ± 0.2) ％],and it increased again when LIDO added (P ＜ 0.05,3 ％ ISO + 100 mg/L LIDO vs.3％ ISO group).Conclusion The results indicate that LIDO could reduce apoptosis of H4 cells induced by ISO by attenuating mitochondrial damage.