欢迎访问《解剖学报》官方网站！今天是
English
色钉菇乙酸乙酯相对SH-SY5Y细胞的保护作用
Protective effects of ethyl acetate extract from Chroogomphis rutilus on SH-SY5Y cells damaged by 1-methyl-4-phenyl pyridine ion
目的 探讨色钉菇乙酸乙酯提取物对1-甲基-4-苯基吡啶离子（MPP+）诱导损伤的多巴胺能神经元的保护作用。 方法 以SH-SY5Y细胞系为对象，首先测定MPP+孵育48 h时的半致死量(IC50)。然后用25mg/L、50mg/L、100mg/L等剂量的乙酸乙酯提取物预先孵育4 h，随后加入1.0mmol/L（IC50值）MPP+孵育48 h。通过MTT法检测细胞活性；Hoechst染色法、Annexin Ⅴ-FITC/PI双染法流式细胞术检测法观察细胞凋亡情况。利用DCFH-DA检测色钉菇乙酸乙酯提取物对细胞内活性氧（ROS）水平的影响。 结果 1.0mmol/L MPP+孵育48 h时达到半致死剂量，MPP+的损伤浓度确定为1.0mmol/L。预先经色钉菇乙酸乙酯相（50mg/L）孵育，可显著提高SH-SY5Y细胞的活力，降低细胞凋亡率。MPP+损伤后ROS水平急剧升高，而25mg/L、50mg/L、100mg/L乙酸乙酯提取物干预后ROS水平下降。 结论 50mg/L色钉菇乙酸乙酯相可显著减轻MPP+对SH-SY5Y细胞的毒性损伤，具有显著的保护作用。
Objective The aim of present study is to observe the protective effects of ethyl acetate extract fromChroogomphis rutiluson dopaminergic neurons damaged by 1-methyl-4-phenyl pyridine ion (MPP+). Methods SH-SY5Y cells (dopaminergic cells), a cell line of human neuroblastoma, were served as the experimental subject. We examined the IC50 of MPP+ at 48 hours. The cells were preincubated with ethyl acetate extract fromChroogomphis rutilusfor 4 hours with 25mg/L, 50mg/L, 100mg/L respectively, then added MPP+ 1.0mmol/L（IC50 value） to incubate for 48 hours. MTT assay, Hoechst 33342 staining and flow cytometric assay were used to detect the survival or apoptosis of SH-SY5Y cells. DCFH-DA was used to detect the reactive oxygen speciess (ROS) level on the cells of ethyl acetate extract fromChroogomphis rutilus. Results 1.0mmol/L MPP+ caused the IC50 of cells at 48 hour, so 1.0mmol/L was used as the lesion concentration. Pretreatment with ethyl acetate extract fromChroogomphis rutilus (50mg/L) significantly increased the cell viability, and reduced the apoptosis rate of cells. After MPP+ lesion,the ROS level increased significantly, and the preincubation with ethyl acetate extract fromChroogomphis rutilus reduced the ROS level induced by MPP+. Conclusion Ethyl acetate extract from Chroogomphis rutilus can significantly protect dopaminergic cells damaged with MPP+.
［1］Dai YCh, Zhou LW, Yang ZhL, et al. A revised checklist of edible fungi in China［J］. Mycosystema, 2010, 29(1): 1-21. (in Chinese)
戴玉成, 周丽伟, 杨祝良, 等. 中国食用菌名录［J］. 菌物学报， 2010,29 (1):1-21.
［2］Feng J, Qin ShL, Hu B, et al.Chemical constituents of fruiting bodies of Chroogomphus rutilus and their biological activities［J］. Mycosystema, 2014, 33(2): 355-364. (in Chiense)
冯建, 秦淑亮, 胡兵, 等. 色钉菇子实体化学成分及其生物活性初探［J］. 菌物学报， 2014， 33(2): 355-364.
［3］Sun H. Studies on the macrofungus Resources in Yanshan mountain of hebei province and antioxidant activity of five kinds of mushrooms［D］. Hebei Normal University, 2011: 25-42. (in Chinese)
孙红. 河北省燕山地区野生高等真菌资源调查及五种子实体抗氧化活性研究［D］. 河北师范大学，2011：25-42.
［4］Zuo L, Motherwell MS. The impact of reactive oxygen species and genetic mitochondrial mutations in Parkinson’s disease［J］. Gene, 2013,532(1): 18-23.
［5］Ciron C, Zheng L, Bobela W, et al. PGC1alpha activity in nigral dopamine neurons determines vulnerability to alphasynuclein［J］. Acta Neuropathol Commun, 2015, 3(1): 16-35.
［6］Zhang XQ, Sun H, Wang LA, et al. Protective effects of crudepolysaccharides from Chroomgomphus rutilus on dopaminergic neurons impaered by MPTP in mice［J］. Mycosystema, 2011, 30(1): 77-84. (in Chinese)
张雪倩, 孙红, 王立安，等. 色钉菇粗多糖对小鼠DA能神经元MPTP损伤的保护作用［J］. 菌物学报， 2011， 30 (1): 77-84.
［7］Zhang XQ, Wang YQ, Wang LA, et al. Protective effects of crude polysaccharides from Chroomgomphus rutilus on SH-SY5Y cells impaired by MPP+［J］. Acta Physiologica Sinica, 2013, 65(2): 210-216. (in Chinese)
张雪倩, 王艳芹, 王立安, 等. 色钉菇粗多糖对MPP+损伤的SH-SY5Y细胞的保护作用［J］. 生理学报， 2013， 65 (2): 210-216.
［8］Gaki GS, Papavassiliou AG. Oxidative stress-induced signaling pathways implicated in the pathogenesis of Parkinson’sdisease［J］. Neuromolecular Med, 2014, 16(2): 217-230.
［9］Jin H, Kanthasamy A, Ghosh A, et al. Mitochondria-targeted antioxidants for treatment of Parkinson’s disease: preclinical and clinical outcomes［J］. Biochim Biophys Acta, 2014, 1842(8): 1282-1294.
［10］Etminan M, Gill SS, Samii A. Intake of vitamin E, vitamin C, and carotenoids and the risk of Parkinson’s disease: a meta-analysis［J］. Lancet Neurol, 2005, 4(6): 362-365.
［11］Weber CA, Ernst ME. Antioxidants, supplements, and Parkinson’s disease［J］. Ann Pharmacother, 2006, 40(5): 935-938.
［12］Li TT, Liang YCh. In vitro models of parkinson disease［J］. Medical Recapitulate, 2013, 19(6): 1003-1006. (in Chinese)
李婷婷, 梁迎春. 帕金森病体外细胞模型［J］. 医学综述, 2013, 19(6): 1003-1006.
［13］Chen L, Zhang LH, Zhang JF, et al. Parkinson’s disease cells models and its experimental research［J］. Health Research, 2013, 33(3): 179-182. (in Chinese)
陈璐, 张丽慧, 张家凤, 等. 帕金森病细胞模型及实验研究［J］. 健康研究, 2013, 33(3): 179-182.
［14］Giordano S, Darley-Usmar V, Zhang J. Autophagy as an essential cellular antioxidant pathway in neurodegenerative disease［J］. Redox Biol, 2014, 2（1）: 82-90.
河北省自然科学基金;河北省科学技术研究与发展计划项目;河北师范大学博士启动基金
/
| 〈 |
|
〉 |
