香椿子多酚通过p38 MAPK通路抑制6-OHDA诱导的神经炎症反应  

Polyphenols from toona sinensis seeds inhibits 6-OHDA-induced neuroinflammation in PC12 cells via the p38 MAPK pathway

作　　者：刘飞[1] 费学超 李侃 庄文欣 吕娥[3] 付文玉[3] 

机构地区：[1]潍坊医学院 [2]潍坊医学院医学研究实验中心,潍坊261053 [3]潍坊医学院组织学与胚胎学教研室,潍坊261053

出　　处：《神经解剖学杂志》2017年第6期665-671,共7页Chinese Journal of Neuroanatomy

基　　金：山东省自然科学基金面上项目(ZR2014HL043);山东省医药卫生科技发展计划(2015WS0063);潍坊医学院大学生科技创新基金(KX2016006,KX2016011),潍坊医学院博士启动基金(2017BSQD26);山东省中医药科技发展计划项目(2015-234,2017-214);潍坊市科学技术发展计划项目(2015WS038)

摘　　要：目的:研究香椿子多酚(PTSS)通过调节p38促分裂原活化蛋白激酶(p38 MAPK)信号通路抑制神经毒素6-羟多巴胺(6-OHDA)所诱导的PC12细胞帕金森病(PD)模型的神经炎症反应。方法:将PC12细胞分为四组:对照组、模型组(6-OHDA 100μmol/L)、PTSS低剂量组(6-OHDA+PTSS 100μmol/L)、PTSS高剂量组(6-OHDA+PTSS 200μmol/L)。倒置显微镜下观察各组PC12细胞形态学的变化;采用细胞计数CCK-8法检测细胞活性;免疫细胞化学染色法和Western Blot检测诱导性一氧化氮合酶(iNOS)、环氧化酶-2(COX-2)、核因子κB p65(NF-κB p65)、p38 MAPK和p-p38 MAPK的表达变化。结果:(1)细胞形态观察显示:与对照组相比,6-OHDA模型组细胞胞体出现空泡,皱缩变形颜色暗淡,部分细胞折光性增高,细胞死亡数目明显增多,细胞聚集成片。PTSS低剂量组,细胞状态明显改善。(2)模型组PC12细胞活力显著降低,而PTSS能有效抑制PC12细胞活力的降低(P<0.05),PTSS低剂量组比高剂量组改善效果更为显著(P<0.05)。(3)与对照组比较,模型组细胞iNOS、COX-2、NF-κB p65、p38 MAPK和p-p38 MAPK表达明显升高,而PTSS组上述分子表达明显降低。结论:PTSS可有效逆转6-OHDA导致的PC12细胞损伤,其机制可能与下调p38 MAPK信号通路从而抑制神经炎症反应有关。Objective： To investigate the effects of polyphenols from toona sinensis seeds （PTSS）, via the p38 mito- gen-activated protein kinase （p38 MAPK） pathway, on suppressing neuroinflammatory responses of PC12 cell model of Parkinson＇s disease （PD） induced by neurotoxin 6-hydroxydopamine （6-OHDA）. Methods： The PC12 cells were di- vided into four groups： control group; model group （6-OHI）A 100 punol/L） ; PTSS low dose group （6-OHDA ＋ PTSS 100 p, mol/L） ; PTSS high dose group （6-OHDA ＋ PTSS 200 p, mol/L）. The morphology of PC12 cells was observed under inverted microscope. The cell viability was detected by CCK-8 method. The expression of inducible nitric oxide synthase （iNOS）, cyclooxygenase-2 （COX-2）, nuclear factor-kappa B p65 （NF-KB p65）, p38 MAPK and p-p38 MAPK wereexamined by immunocytochemical staining and Western Blot. Results： （ 1 ） Compared with the control group, the cells in the model group demonstrated vacuoles, deformation or aggregation. The light refraction of some cells was enhanced and the dead cells were significantly inereased, and the cells aggregated. The morphology of cells in PTSS low dose group was obviously improved. （2） The cell viability was significantly decreased in the model group; PTSS could signif- icantly suppress the decreasing viability, as in the two PTSS groups （P 〈 0. 05）. This improving effect of PTSS on cell viability in PTSS low dose group was higher than that in the high dose group （ P 〈 0.05 ）. （3） Compared with the con- trol group, the expression of iNOS, COX-2, NF-κB p65, p38 MAPK and p-p38 MAPK in the model group was increased markedly. PTSS could effectively reduce the expression of these molecules. Conclusion： PTSS can effectively reverse the damage of PC12 cells induced by 6-OHDA. The mechanism may be related to restraining the p38 MAPK signaling pathway whereby neuroinilammatory responses were inhibited.

关 键 词：香椿子多酚 6-羟多巴胺 帕金森病 P38 MAPK 神经炎症 

分 类 号：R285.5[医药卫生—中药学]