藁本内酯对帕金森病细胞模型的保护作用及其机制研究  

Protective effect of ligustilide in Parkinson’s disease cell model and its mechanism

作　　者：汤灿辉[1] 王梦禅[2] TANG Can-hui;WANG Meng-chan(Department of Pharmacy,Jiangxi College of Traditional Chinese Medicine,Fuzhou 344000,Jiangxi Province,China;Schoolof Pharmacy,Chongqing Three Gorges Medical College,Chongqing 404120,China)

机构地区：[1]江西中医药高等专科学校药学系江西,江西抚州344000 [2]重庆三峡医药高等专科学校药学院,重庆404120

出　　处：《中国临床药理学杂志》2023年第6期795-799,共5页The Chinese Journal of Clinical Pharmacology

摘　　要：目的研究藁本内酯对帕金森病细胞模型的保护作用及其作用机制。方法取对数生长期的SK-N-SH细胞使用200μmol·L^(-1)的MPP+处理48 h,构建帕金森病细胞模型;以正常培养基培养的细胞作为对照组。藁本内酯组加入终浓度为30.0μmol·L^(-1)藁本内酯进行培养;P79350组加入30.0μmol·L^(-1)的藁本内酯后,实验结束前1 h加入50μmol·L^(-1)的P79350处理细胞;对照组和MPP+组加入不含药物的培养基培养。用细胞计数试剂盒-8(CCK-8)检测细胞活力;以流式细胞术检测细胞周期情况和细胞凋亡率;以蛋白质印迹法检测p38丝裂原活化蛋白激酶/c-Jun氨基末端激酶(p38 MAPK/JNK)信号通路和增殖、凋亡相关蛋白的表达水平。结果对照组、MPP+组和10.0、20.0、30.0、50.0、100.0和200.0μmol·L^(-1)藁本内酯组的细胞存活率分别为(100.00±1.91)%、(35.63±3.40)%、(49.06±3.78)%、(54.08±6.63)%、(91.61±4.43)%、(79.87±6.15)%、(64.73±7.64)%和(42.99±6.09)%。藁本内酯可明显抑制MPP+诱导引起的p38 MAPK/JNK信号通路的激活,并明显抑制MPP+组细胞G0/G1期转变。对照组、MPP+组、藁本内酯组和P79350组细胞凋亡率分别为(4.66±0.68)%、(21.22±1.98)%、(10.43±0.96)%和(15.32±1.41)%。与MPP+组细胞相比,藁本内酯作用后可明显促进细胞中Ki-67、增殖细胞核抗原(PCNA)和B淋巴细胞瘤-2(Bcl-2)蛋白的表达,而明显抑制Bcl-2相关X(Bax)蛋白和裂解的胱天蛋白酶-3(Cl-caspase-3)蛋白的表达,差异均有统计学意义(均P<0.05)。结论藁本内酯对帕金森病细胞模型具有保护作用,可增强细胞活力,抑制细胞凋亡,主要与抑制p38 MAPK/JNK信号通路的激活有关。Objective To study the protective effect and its mechanism of ligustilide in Parkinson’s disease cell model.Methods SK-N-SH cells in logarithmic phase were treated with 200 mol·L^(-1)MPP++for48 h to establish Parkinson’s cell model.The cells cultured in norma medium were used as control.Ligustilide group was cultured with 30.0μmol·L^(-1)ligustilide.After 30.0μmol·L^(-1)ligustilide was added into the P79350 group,50μmol·L^(-1)P79350 was added to treat the cells a1 h before the end of the experiment.The control group and MPP+group were cultured in drug-free medium.The cell viability was detected by cell counting kit-8(CCK-8).Cell cycle and apoptosis rate were detected by flow cytometry.The expression levels of p38 mitogen activated protein kinase/c-Jun N-terminal kinase(p38 MAPK/JNK)signaling pathway related proteins and proliferation and apoptosis related proteins were detected by Western Blotting.Results The cell viability of the control,MPP+and 10.0,20.0,30.0,50.0,100.0 and 200.0μmol·L^(-1)ligustilide groups were(100.00±1.91)%,(35.63±3.40)%,(49.06±3.78)%,(54.08±6.63)%,(91.61±4.43)%,(79.87±6.15)%,(64.73±7.64)%and(42.99±6.09)%,respectively.Ligustilide significantly inhibited the MPP+induced activation of p38 MAPK/JNK signaling pathway and significantly inhibited the G0/G1 phase transformation in MPP+group.The apoptosis rates in the control,MPP+,ligustilide,and P79350 groups were(4.66±0.68)%,(21.22±1.98)%,(10.43±0.96)%and(15.32±1.41)%,respectively.Compared with the cells in the MPP+group,ligustilide significantly increased the expression of Ki67,proliferating cell nuclear antigen(PCNA)and B-cell lymphoma-2(Bcl-2)proteins,while significantly inhibited the expression of Bcl-2 associated X(Bax)and cleaved cysteine aspartate proteinase-3(Cl-caspase-3)proteins,and the difference was statistically significant(P<0.05).Conclusion Ligustilide has protective effect on Parkinson’s model cells;it can enhance cell viability and inhibit apoptosis,which is related to the activation of p38 MAPK/JNK signa

关 键 词：藁本内酯 帕金森病 增殖 凋亡 p38丝裂原活化蛋白激酶/c-Jun氨基末端激酶(p38 MAPK/JNK)信号通路 

分 类 号：R28[医药卫生—中药学]