氯胺酮、利多卡因对分化的PC12细胞内游离钙浓度的影响  

Effects of ketamine and lidocaine on intracellular free calcium concentration in differentiated PC12 cells

作　　者：张倬宁 刘莉莉 周晓平 杨百川 孙昕培 韩雨佳 冷恒芳 张云 于剑锋[1] ZHANG Zhuoning;LIU Lili;ZHOU Xiaoping;YANG Baichuan;SUN Xinpei;HAN Yujia;LENG Hengfang;ZHANG Yun;YU Jianfeng(Department of Anesthesia,Weifang Medical University,Weifang 261053,China;Experimental Center for Medical Research,Weifang Medical University)

机构地区：[1]潍坊医学院麻醉学院,山东潍坊261053 [2]潍坊医学院医学研究实验中心

出　　处：《潍坊医学院学报》2021年第5期383-385,共3页Acta Academiae Medicinae Weifang

基　　金：国家自然科学基金(项目编号:81241041);潍坊医学院大学生科技创新基金(项目编号:S202010438071)。

摘　　要：目的探讨不同浓度的氯胺酮、利多卡因对PC12细胞内游离钙的影响及其作用机制。方法将经NGF诱导分化的神经元样PC12细胞接种于24孔板内,培养8~12h后,细胞随机分为氯胺酮处理组(K组):分别为10^(-2)、10^(-3)、10^(-4)、10^(-5)、10^(-6)倍比稀释的氯胺酮;利多卡因处理组(L组):分别为10^(-2)、10^(-3)、10^(-4)、10^(-5)、10^(-6)、10^(-7)、10^(-8)倍比稀释的利多卡因。用Fura 2-AM染料作为Ca^(2+)指示剂,37℃下共孵育40min,人工脑脊液洗涤后,选定多个细胞分别观察加入染料后340/380nm激发荧光强度比值的变化,代表细胞内钙离子浓度([Ca^(2+)]i)的变化。结果10^(-2)~10^(-6)倍比稀释的氯胺酮或10^(-2)~10^(-8)倍比稀释的利多卡因使分化的PC12细胞340/380nm激发荧光强度比值明显下降。结论一定浓度的氯胺酮或利多卡因明显降低分化的PC12细胞内钙离子浓度([Ca^(2+)]i),可能分别与NMDA受体和钠通道被抑制有关。Objective To investigate the effects of different concentrations of ketamine and lidocaine on intracellular free calcium in PC12 cells and its mechanism.Methods The neuron-like PC12 cells induced by NGF were planted in 24-well plates and incubated for 8~12h,and then were randomly divided into K groups diluted to 10^(-2),10^(-3),10^(-4),10^(-5)10^(-6) folds of ketamine,and L groups diluted to 10^(-2),10^(-3),10^(-4),10^(-5),10^(-6),10^(-7),10^(-8) folds of lidocaine.The cells were co-incubated with Fura 2-AM dye as indcaitor of Ca^(2+)for 40min at 37℃.Excess dye was washed by artificial cerebrospinal fluid(ACSF).Several cells were selected and measured the changes in the ratio of fluorescence intensities excited at 340/380nm after adding dye which represents the change in[Ca^(2+)]i.Results Ketamine diluted to 10^(-2)~10^(-6) folds or lidocaine diluted to 10^(-2)~10^(-8) folds significantly decreased 340/380nm fluorescence intensity ratio of differentiated PC12 cells.Conclusion A certain concentration of ketamine or lidocaine can significantly reduce the[Ca^(2+)]i of differentiated PC12 cells,which may be related to the inhibition of NMDA receptor and sodium channel respectively.

关 键 词：氯胺酮 利多卡因 PC12细胞 钙离子浓度 Fura 2-AM 

分 类 号：R614[医药卫生—麻醉学]