[1]宋俊科,张 雯,张 雪,等.丹酚酸D对MPP+损伤SH-SY5Y细胞线粒体功能和生物合成的影响[J].中国药理学通报,2018,(09):1211-1218.[doi:10.3969/j.issn.1001-1978.2018.09.007]
 SONG Jun-ke,ZHANG Wen,ZHANG Xue,et al.Effects of salvianolic acid D on mitochondrial function and biosynthesis in SH-SY5Y cells after MPP+ injury[J].Chinese Pharmacological Bulletin,2018,(09):1211-1218.[doi:10.3969/j.issn.1001-1978.2018.09.007]
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丹酚酸D对MPP+损伤SH-SY5Y细胞线粒体功能和生物合成的影响()
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《中国药理学通报》[ISSN:/CN:]

卷:
期数:
2018年09期
页码:
1211-1218
栏目:
论著
出版日期:
2018-08-26

文章信息/Info

Title:
Effects of salvianolic acid D on mitochondrial function and biosynthesis in SH-SY5Y cells after MPP+ injury
文章编号:
1001-1978(2018)09-1211-08
作者:
宋俊科张 雯张 雪王金华杨海光杜冠华
中国医学科学院北京协和医学院药物研究所,天然药物活性物质与功能国家重点实验室,药物靶点研究与新药筛选北京市重点实验室,北京 100050
Author(s):
SONG Jun-ke ZHANG Wen ZHANG Xue WANG Jin-hua YANG Hai-guang DU Guan-hua
State Key Lab of Bioactive Substances and Functions of Natural Medicines, Beijing Key Lab of Drug Target and Screening Research, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China
关键词:
丹酚酸D MPP+ SH-SY5Y 线粒体功能 线粒体生物合成 PGC-1α
Keywords:
salvianolic acid D MPP+ SH-SY5Y mitochondrial function mitochondrial biosynthesis PGC-1α
分类号:
R284.1; R329.24; R329.25; R342.2; R977.6
DOI:
10.3969/j.issn.1001-1978.2018.09.007
文献标志码:
A
摘要:
目的 研究丹酚酸D(salvianolic acid D, SalD)对MPP+损伤SH-SY5Y细胞线粒体功能和生物合成的影响及机制。方法 采用MPP+损伤SH-SY5Y细胞模型,MTT法检测MPP+对细胞的毒性作用,MTT和LDH法考察SalD对SH-SY5Y细胞活力的影响,AO/EB法检测细胞凋亡。应用DCFH-DA和MitoSOX分别检测细胞ROS及线粒体超氧化物水平,通过测定细胞内ATP水平以及线粒体膜电位变化,考察线粒体功能。qPCR检测MPP+损伤细胞后,PGC-1α及其下游调控基因NRF1和TFAM的mRNA水平,Western blot及免疫荧光测定细胞中PGC-1α、NRF1和TFAM蛋白含量。结果 MPP+可损伤SH-SY5Y细胞,导致细胞存活率明显降低为51.34%。0.1、1、5 μmol·L-1 SalD和5 mmol·L-1 NAC能够减轻MPP+诱导的SH-SY5Y细胞损伤及LDH释放,其细胞存活率分别增加到67.98%、71.79%、76.91%、77.55%。并且,SalD能减少MPP+诱导的细胞内ROS及线粒体超氧化物的升高,降低线粒体膜电位,改善线粒体功能。同时,SalD能够明显抑制MPP+损伤导致的PGC-1α、NRF1、TFAM mRNA转录及表达降低。结论 SalD能够抑制MPP+诱导的SH-SY5Y细胞损伤,保护线粒体功能和线粒体生物合成。
Abstract:
Aim To investigate the effects of salvianolic acid D(SalD)on mitochondrial function and biosynthesis in SH-SY5Y cells after MPP+ injury and the possible mechanisms. Methods The cell model was established by MPP+ injury in SH-SY5Y cells. The cytotoxicity of MPP+ was detected by MTT assay. The effects of SalD on viability of SH-SY5Y cells were examined by MTT and LDH assay. The apoptosis of SH-SY5Y cells was detected by AO/EB assay. The levels of ROS and mitochondrial superoxide were determined using DCFH-DA and MitoSOX probes, respectively. Mitochondrial function was examined by measuring ATP level and mitochondrial membrane potential. The levels of PGC-1α and its downstream regulatory genes NRF1 and TFAM mRNA were detected by qPCR. The protein levels of PGC-1α, NRF1 and TFAM in cells were detected by Western blot and immunofluorescence assays. Results MPP+ injury resulted in a significant reduction of cell viability to 51.34%. 0.1, 1, 5 μmol·L-1 SalD and 5 mmol·L-1 NAC could reduce MPP+-induced SH-SY5Y cell injury and LDH release. The cell viability increased to 67.98%, 71.79%, 76.91% and 77.55%, respectively. Moreover, SalD could reduce the increase of intracellular ROS and mitochondrial superoxide induced by MPP+, decrease mitochondrial membrane potential and improve mitochondrial function. SalD also significantly increased both the transcription and expression levels of PGC-1α, NRF1 and TFAM. Conclusion SalD could inhibit MPP+-induced SH-SY5Y cell injury and improve mitochondrial function and mitochondrial biosynthesis.

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备注/Memo

备注/Memo:
收稿日期:2018-05-20,修回日期:2018-06-24
基金项目:国家自然科学基金资助项目(No 81603100); 中国医学科学院医学与健康科技创新工程(No 2017-I2M-1-010)
作者简介:宋俊科(1988-),男,博士,助理研究员,研究方向:神经药理学与新药发现,E-mail:smilejunke@imm.ac.cn;
杜冠华(1956-),男,博士,研究员,博士生导师,研究方向:神经药理学、心脑血管药理学与新药发现,通讯作者,E-mail:dugh@imm.ac.cn
更新日期/Last Update: 2018-08-26