川芎对神经细胞保护作用活性成分的研究  

Study on the Active Components of Ligusticum Chuanxiong for Neuroprotection

作　　者：王敏[1] 姚明江[1] 刘建勋[1] 吴昱杰 王攀 任钧国[1] 范晓迪[1] WANG Min;YAO Ming-jiang;LIU Jian-xun;WU Yu-jie;WANG Pan;REN Jun-guo;FAN Xiao-di(Research Center,Xiyuan Hospital,Chinma Academy of Chinese Medical Seciences,Beijing 100091;Beijing Universityof Chinese Medicine,Beijing 100091)

机构地区：[1]中国中医科学院,西苑医院基础医学研究所,北京100091 [2]北京中医药大学,北京100091

出　　处：《世界中西医结合杂志》2021年第5期793-798,共6页World Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金资助项目(81873041,82030124)。

摘　　要：目的对川芎中主要的几种成分神经保护的活性进行研究。方法采用CCK-8法检测洋川芎内酯Ⅰ(SEI)、洋川芎内酯H(SEH)、洋川芎内酯A(SEA)、阿魏酸(FA)、藁本内酯(LIG)5种成分对SH-SY5Y细胞毒性;CCK-8法检测不同缺氧、复氧时间对细胞存活率的影响,建立氧糖剥夺/复氧复糖(oxygen glucose deprivation-reoxygenation,OGD-R)为模型;分别检测化合物给药后对OGD-R损伤模型细胞存活率的影响、对缺氧液和复氧液中乳酸脱氢酶(LDH)释放的影响以及应用流式细胞术检测细胞内活性氧ROS的变化。结果SEI 100μmol/L、SEH 50μmol/L、SEA 200μmol/L、LIG 25μmol/L、FA 100μmol/L及以下浓度对细胞无明显毒性,可用于进一步实验研究;细胞缺氧缺糖2 h,复氧复糖24 h可作为神经保护药物活性筛选的条件;与模型组比较,SEI(50μmol/L)、FA(50、25μmol/L)、LIG(12.5、6.25μmol/L)均能显著抑制模型损伤细胞存活率的下降;SEI(50、25μmol/L)、FA(50μmol/L)、LIG(6.25μmol/L)均能显著降低LDH释放,FA 25μmol/L、LIG 12.5μmol/L对LDH无显著性影响;SEI(50、25μmol/L)、LIG(12.5、6.25μmol/L)均能降低细胞内ROS。结论SEI、LIG对细胞存活率、LDH活力、细胞内ROS均有改善作用,其中SEI对LDH抑制作用更明显且呈剂量依赖,SEI和LIG在神经保护中的药理作用需进一步研究。Objective To screen components with neuroprotective activities from Ligusticum Chuanxiong.Methods The CCK-8 method was used to detect the cytotoxicity of SenkyunolideⅠ(SEI),Senkyunolide H(SEH),Senkyunolide A(SEA),the Ferulic Acid(FA),and the Ligustilide(LIG)from Ligusticum Chuanxiong.To establish oxygen glucose deprivation-reoxygenation(OGD-R)model,SH-SY5 Y cells were exposed to different hypoxia and reoxygenation time and cell viability was detected by CCK-8.Then the release of lactate dehydrogenase(LDH)in hypoxia and reoxygenation solutions was detected and the intracellular reactive oxygen species(ROS)were evaluated by flow cytometry after treatment with the compounds.Results SEI 100μmol/L,SEH 50μmol/L,SEA 200μmol/L,LIG 25μmol/L,FA 100μmol/L had no obvious toxicity to cells and could be used for following experiments.Hypoxia and Glucose deprivation for 2 hours and reoxygenation and complex sugar for 24 hours can be used as conditions for screening neuroprotective drug activity.Compared with the model group,SEI(50μmol/L),FA(50,25μmol/L),LIG(12.5,6.25μmol/L)could significantly inhibit the decrease of cell viability of model injured cells,respectively.Besides,SEI(50,25μmol/L),FA(50μmol/L),LIG(6.25μmol/L)treatment reduced LDH significantly,while FA 25μmol/L and LIG 12.5μmol/L showed no obvious effect.Furthermore,SEI(50,25μmol/L)and LIG(12.5,6.25μmol/L)could reduce intracellular ROS.Conclusion The results showed that SEI and LIG could improve cell survival rate,LDH,and intracellular ROS.Among them,SEI had a more obvious inhibitory effect on LDH and was dose-dependent.The pharmacological effects of SEI and LIG in neuroprotection are worthy of attention and further research.

关 键 词：川芎 洋川芎内酯Ⅰ 藁本内酯 阿魏酸 氧糖剥夺 SH-SY5Y 

分 类 号：R285.6[医药卫生—中药学]