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作 者:高华[1] 隋雷鸣[2] 谷利[3] 杨巍巍[3] 鲁玲玲[3] 赵莎莎[3] 杨慧[3]
机构地区:[1]北京市神经外科研究所,北京100050 [2]首都医科大学医学实验与测试中心 [3]首都医科大学神经生物学系 神经再生修复研究重点实验室 神经变性病教育部重点实验室,北京100069
出 处:《基础医学与临床》2013年第11期1430-1434,共5页Basic and Clinical Medicine
基 金:国家重点基础研究发展计划(2011CB504102;2012CB722407);国家自然科学基金(30970940);北京市教育委员会科技发展计划重点项目(KZ201010025022);北京市自然基金面上项目(5102012);北京市高校人才强教资助PHR(200907113);北京市神经再生修复研究重点实验室开放课题(2011SJZS02);神经变性病教育部重点实验室开放课题(2011SJBX03)
摘 要:目的观察过表达DJ-1蛋白能否保护MN9D细胞抵抗鱼藤酮所致的线粒体损伤和细胞凋亡。方法在MN9D细胞中分别过表达野生型DJ-1(WT)和L166P突变型DJ-1(L166P),随后给予鱼藤酮处理。MTT法观察细胞活力、JC-1染色观察线粒体膜电位(ΔΨm)以及流式细胞仪AnnexinⅤ+PI染色和TUNEL染色观察凋亡。结果单纯鱼藤酮处理组细胞活力下降48.2%±6.4%,而过表达WT时,加入鱼藤酮后细胞活力仅下降22.0%±3.7%(P<0.05),过表达L166P组跟单纯鱼藤酮组无显著性差异;鱼藤酮引起MN9D细胞ΔΨm下降,过表达WT则可以部分恢复ΔΨm,L166P无此作用;过表达WT-DJ-1组凋亡率分别为5.4%和9.7%,较单纯鱼藤酮处理明显下降(P<0.05),而转染L166P细胞凋亡率跟鱼藤酮组无差异,TUNEL实验证实了同样的结果。结论过表达野生型DJ-1能缓解鱼藤酮对MN9D细胞的损伤,具有线粒体保护功能,减少细胞凋亡的发生,而突变型L166P则无此作用。Objective To observe whether DJ-1 can relieve the mitochondrial dysfunctions and cell apoptosis in MN9D cellafter rotenone exposure. Methods MN9D cell line was exposed to rotenone ( 100 nmol/L) for 24 h after being trans- fected with human mutant (L166P) or wild-type (WT) DJ-1. Cells were divided into Co, WT-DJ-1 (WT), L166P-DJ-1 (L166P) group. Cell viability was measuled by MTr, and mitochondrial membrane potential (△ψm) by JC-1 and cell apoptosis by Annexin V + PI and TUNLE staining. Results Cell viability of Co group reduced to48.2% ±6. 4% , and WT group was 78± 3.7% (P 〈 0. 05 ), there was no statistical difference between Co group and L166P group. Overexpression of WT-DJ-1 partly rescued △ψm which was reduced after rotenone expo- sure (P 〈0. 05) , however, overexpression of L166P exacerbated the reduce of A^m. And cell death in WT group was 5.4% and 9.7% after rotenone exposure for 12h or 24h respectively, accordingly 10. 9% and 23.2% in Co group (P 〈0. 05 ). There was no statistic difference between Co group and L166P group. Conclusions Overex- pression WT-DJ-1 may relieve the cell injury after rotenone exposure and dull the mitochondrial dysfunction. How- ever, L166P has not the protective effect after rotenone exposure.
关 键 词:帕金森病 DJ-1 鱼藤酮 凋亡 线粒体功能障碍
分 类 号:R742.5[医药卫生—神经病学与精神病学]
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