香椿子多酚通过JNK通路抑制6-羟多巴胺诱导PC12细胞炎症损伤
|
摘要
目的研究香椿子多酚(PTSS)通过c-Jun氨基末端激酶(JNK)通路调节神经毒素6-羟多巴胺(OHDA)诱导的PC12细胞炎症损伤。方法将PC12细胞分为对照组、模型组(6-OHDA 100μmol/L)、PTSS组(6-OHDA+PTSS 100μmol/L)。在倒置显微镜下观察各组PC12细胞的形态学变化;采用细胞计数试剂盒(CCK)-8检测细胞活性;免疫细胞化学染色法和Western印迹检测JNK、p-JNK、炎症因子白细胞介素(IL)-1β和IL-6的表达变化。结果细胞形态观察结果显示,对照组细胞数量多,形态良好。与对照组比较,6-OHDA作用24 h后,模型组PC12细胞数量明显减少,聚集成团。而PTSS作用12 h后,PTSS组细胞数量较模型组多,形态明显好于模型组。CCK-8法显示,与对照组相比,模型组细胞活力显著降低(P<0.05),PTSS组细胞活力明显上升(P<0.05)。与对照组比较,模型组PC12细胞JNK、p-JNK、IL-1β和IL-6的含量均明显提高(P<0.05),而PTSS组上述蛋白及因子的含量均显著下降(P<0.05)。结论 PTSS可通过抑制JNK通路的信号分子及其下游炎症因子对抗6-OHDA诱导的PC12细胞的神经炎症反应。
Objective To investigate the anti-neuroinflammatory responses of polyphenols from toonasinensis seeds(PTSS) on 6-hydroxydopamine(OHDA)-induced PC12 cells model of Parkinson's disease(PD) via the c-Jun N-terminal kinase(JNK) pathway. Methods PC12 cells were divided into control, model(6-OHDA 100 μmol/L), and PTSS(6-OHDA+PTSS 100 μmol/L) groups. The morphological changes of PC12 cells in each group were observed under an inverted microscope. Cell viability was determined by CCK-8 method. The expressions of JNK, p-JNK, IL-1β and IL-6 were detected by immunocytochemistry staining and Western blot.Results Cells in control group were abundant cytoplasm and distributed evenly. Compared with those of control group, the PC12 cells in model group were reduced obviously and deformed evidently after 6-OHDA treatment for 24 h. The number of cells after PTSS treatment for 12 h was more than that of model group, and the morphology of cells was obviously improved. Compared with that of control group, the cell viability of model group was markedly reduced(P<0.05). PTSS treatment for 12 h markedly suppressed the reducing viability(P<0.05). Compared with that of control group, the expressions of IL-1β, IL-6, JNK and p-JNK in model group were obviously increased(P<0.05), while the expressions of the above proteins and factors in PTSS group were significantly decreased(P<0.05). Conclusions PTSS could counteract 6-OHDA-induced neuroinflammation of PC12 cells by inhibiting the signaling molecules of the JNK pathway and its downstream inflammatory factors.
Objective To investigate the anti-neuroinflammatory responses of polyphenols from toonasinensis seeds(PTSS) on 6-hydroxydopamine(OHDA)-induced PC12 cells model of Parkinson's disease(PD) via the c-Jun N-terminal kinase(JNK) pathway. Methods PC12 cells were divided into control, model(6-OHDA 100 μmol/L), and PTSS(6-OHDA+PTSS 100 μmol/L) groups. The morphological changes of PC12 cells in each group were observed under an inverted microscope. Cell viability was determined by CCK-8 method. The expressions of JNK, p-JNK, IL-1β and IL-6 were detected by immunocytochemistry staining and Western blot.Results Cells in control group were abundant cytoplasm and distributed evenly. Compared with those of control group, the PC12 cells in model group were reduced obviously and deformed evidently after 6-OHDA treatment for 24 h. The number of cells after PTSS treatment for 12 h was more than that of model group, and the morphology of cells was obviously improved. Compared with that of control group, the cell viability of model group was markedly reduced(P<0.05). PTSS treatment for 12 h markedly suppressed the reducing viability(P<0.05). Compared with that of control group, the expressions of IL-1β, IL-6, JNK and p-JNK in model group were obviously increased(P<0.05), while the expressions of the above proteins and factors in PTSS group were significantly decreased(P<0.05). Conclusions PTSS could counteract 6-OHDA-induced neuroinflammation of PC12 cells by inhibiting the signaling molecules of the JNK pathway and its downstream inflammatory factors.
引文
1 张丽娟,邵海涛,王跃秀,等.帕金森病研究进展[J].生命科学,2014;26(6):560-70.
2 Yan J,Fu Q,Cheng L,et al.Inflammatory response in Parkinson′ s disease[J].Mol Med Rep,2014;10(5):2223-33.
3 Deleidi M,Gasser T.The role of inflammation in sporadic and familial Parkinson′s disease [J].Cell Mol Life Sci,2013;70(22):4259-73.
4 Liu Y,Song Y,Zhu X.MicroRNA-181a regulates apoptosis and autophagy process in Parkinson′s disease by inhibiting p38 mitogen-activated protein kinase (MAPK)/c-Jun N-terminal kinases (JNK) signaling pathways[J].Med Sci Monit,2017;23:1597-606.
5 Huang Q,Du X,He X,et al.JNK-mediated activation of ATF2 contributes to dopaminergic neurodegeneration in the MPTP mouse model of Parkinson′s disease[J].Exp Neurol,2016;277:296-304.
6 李晓健,庄文欣,吕娥,等.香椿子多酚对6-OHDA所致帕金森病大鼠模型的抗炎和神经保护作用[J].神经解剖学杂志,2016;32(5):653-9.
7 刘金城,庄文欣,李锋杰,等.香椿子多酚提取物对6-OHDA诱导的PC12细胞损伤的保护作用[J].神经解剖学杂志,2016;32(1):31-6.
8 刘飞,费学超,李侃,等.香椿子多酚通过p38 MAPK通路抑制6-OHDA诱导的神经炎症反应[J].神经解剖学杂志,2017;33(6):665-71.
9 成晓华,刘斌,马原源,等.咪多吡对帕金森病模型大鼠黑质炎性因子TNF-α、IL-1β、IFN-γ表达的影响[J].免疫学杂志,2014;30(8):671-3.
10 张田,魏子峰,秦丽娟,等.罗格列酮对帕金森病模型小鼠中脑黑质iNOS和 IL-6 表达的抑制作用及其意义[J].吉林大学学报(医学版),2012;38(6):1124-8,1252-3.
11 Jing H,Wang S,Wang M,et al.Isobavachalcone attenuates MPTP-induced Parkinson′s disease in mice by inhibition of microglial activation through NF-κB pathway[J].PLoS One,2017;12(1):e0169560.
12 Ali T,Badshah H,Kim TH,et al.Melatonin attenuates D-galactose-induced memory impairment,neuroinflammation and neurodegeneration via RAGE/NF-κB/JNK signaling pathway in aging mouse model[J].J Pineal Res,2015;58(1):71-85.
13 关榆根,张建鹏.JNK信号通路的激活在帕金森病发病机制中的作用[J].生物技术通讯,2012;23(6):874-8.
14 Zhang S,Gui XH,Huang LP,et al.Neuroprotective effects of β-asarone against 6-hydroxy dopamine-induced parkinsonism via JNK/Bcl-2/Beclin-1 pathway[J].Mol Neurobiol,2016;53(1):83-94.
15 Xu J,Gao X,Yang C.Resolvin D1 Attenuates Mpp+-induced Parkinson disease via inhibiting inflammation in PC12 cells[J].Med Sci Monit,2017;23:2684-91.
16 Lee Y,Chun HJ,Lee KM,et al.Silibinin suppresses astroglial activation in a mouse model of acute Parkinson′s disease by modulating the ERK and JNK signaling pathways[J].Brain Res,2015;1627:233-42.
17 刘迪,李彦秋,李义清,等.香椿子有效组分提取与生物活性研究进展[J].山东化工,2018;47(2):55-6.
18 张京芳,王冬梅,张强.香椿叶抗脂质过氧化物的分离及抗氧化特性[J].农业工程学报,2009;25(1):285-9.
19 马迪,陈超,吴薇,等.香椿子正丁醇提取物对脑缺血再灌注小鼠的神经保护作用及其机制研究[J].解放军医学杂志,2013;38(1):15-8.
2 Yan J,Fu Q,Cheng L,et al.Inflammatory response in Parkinson′ s disease[J].Mol Med Rep,2014;10(5):2223-33.
3 Deleidi M,Gasser T.The role of inflammation in sporadic and familial Parkinson′s disease [J].Cell Mol Life Sci,2013;70(22):4259-73.
4 Liu Y,Song Y,Zhu X.MicroRNA-181a regulates apoptosis and autophagy process in Parkinson′s disease by inhibiting p38 mitogen-activated protein kinase (MAPK)/c-Jun N-terminal kinases (JNK) signaling pathways[J].Med Sci Monit,2017;23:1597-606.
5 Huang Q,Du X,He X,et al.JNK-mediated activation of ATF2 contributes to dopaminergic neurodegeneration in the MPTP mouse model of Parkinson′s disease[J].Exp Neurol,2016;277:296-304.
6 李晓健,庄文欣,吕娥,等.香椿子多酚对6-OHDA所致帕金森病大鼠模型的抗炎和神经保护作用[J].神经解剖学杂志,2016;32(5):653-9.
7 刘金城,庄文欣,李锋杰,等.香椿子多酚提取物对6-OHDA诱导的PC12细胞损伤的保护作用[J].神经解剖学杂志,2016;32(1):31-6.
8 刘飞,费学超,李侃,等.香椿子多酚通过p38 MAPK通路抑制6-OHDA诱导的神经炎症反应[J].神经解剖学杂志,2017;33(6):665-71.
9 成晓华,刘斌,马原源,等.咪多吡对帕金森病模型大鼠黑质炎性因子TNF-α、IL-1β、IFN-γ表达的影响[J].免疫学杂志,2014;30(8):671-3.
10 张田,魏子峰,秦丽娟,等.罗格列酮对帕金森病模型小鼠中脑黑质iNOS和 IL-6 表达的抑制作用及其意义[J].吉林大学学报(医学版),2012;38(6):1124-8,1252-3.
11 Jing H,Wang S,Wang M,et al.Isobavachalcone attenuates MPTP-induced Parkinson′s disease in mice by inhibition of microglial activation through NF-κB pathway[J].PLoS One,2017;12(1):e0169560.
12 Ali T,Badshah H,Kim TH,et al.Melatonin attenuates D-galactose-induced memory impairment,neuroinflammation and neurodegeneration via RAGE/NF-κB/JNK signaling pathway in aging mouse model[J].J Pineal Res,2015;58(1):71-85.
13 关榆根,张建鹏.JNK信号通路的激活在帕金森病发病机制中的作用[J].生物技术通讯,2012;23(6):874-8.
14 Zhang S,Gui XH,Huang LP,et al.Neuroprotective effects of β-asarone against 6-hydroxy dopamine-induced parkinsonism via JNK/Bcl-2/Beclin-1 pathway[J].Mol Neurobiol,2016;53(1):83-94.
15 Xu J,Gao X,Yang C.Resolvin D1 Attenuates Mpp+-induced Parkinson disease via inhibiting inflammation in PC12 cells[J].Med Sci Monit,2017;23:2684-91.
16 Lee Y,Chun HJ,Lee KM,et al.Silibinin suppresses astroglial activation in a mouse model of acute Parkinson′s disease by modulating the ERK and JNK signaling pathways[J].Brain Res,2015;1627:233-42.
17 刘迪,李彦秋,李义清,等.香椿子有效组分提取与生物活性研究进展[J].山东化工,2018;47(2):55-6.
18 张京芳,王冬梅,张强.香椿叶抗脂质过氧化物的分离及抗氧化特性[J].农业工程学报,2009;25(1):285-9.
19 马迪,陈超,吴薇,等.香椿子正丁醇提取物对脑缺血再灌注小鼠的神经保护作用及其机制研究[J].解放军医学杂志,2013;38(1):15-8.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |