β-asarone对Aβ1-42激活的ACM所致PC12细胞损伤的保护作用研究

doi:10.12092/j.issn.1009-2501.2018.06.002

中国临床药理学与治疗学 ›› 2018, Vol. 23 ›› Issue (6): 608-613.doi: 10.12092/j.issn.1009-2501.2018.06.002

β-asarone对Aβ_1-42激活的ACM所致PC12细胞损伤的保护作用研究

何潆¹，包玉婷²，姚莹³，宣玲²，杨元宵³

¹杭州市红十字会医院，杭州 310003，浙江；²浙江中医药大学，杭州 310053，浙江；³杭州医学院，杭州 310053，浙江

收稿日期:2017-09-18 修回日期:2017-10-11 出版日期:2018-06-26 发布日期:2018-06-19
通讯作者: 杨元宵，女，博士，副教授，主要从事中药药理与新产品开发研究。 Tel：0571-87692678 E-mail：yyx104475@163.com
作者简介:何潆，女，硕士研究生，中药师，主要从事中药药理与新产品开发研究。 Tel：0571-56108628 E-mail:yilliahe2013@163.com
基金资助:
国家自然科学基金（81403128）；浙江省医药卫生科技计划（2015KYA062）

Protective effects of β-asarone on PC12 cells injury induced by Aβ_1-42 activated ACM

HE Ying¹, BAO Yuting ², YAO Ying³, XUAN Ling², YANG Yuanxiao³

¹Hangzhou Red Cross Hospital, Hangzhou 310003, Zhejiang, China; ²Zhejiang Chinese Medical University, Hangzhou 310053, Zhejiang, China; ³Hangzhou Medical Collage, Hangzhou 310053, Zhejiang, China

Received:2017-09-18 Revised:2017-10-11 Online:2018-06-26 Published:2018-06-19

摘要/Abstract

摘要：

目的：探讨不同浓度β-淀粉样蛋白（Aβ1-42）激活的星形胶质细胞条件培养液（ACM）对PC12细胞存活率及脑源性神经营养因子（BDNF）、乙酰胆碱酯酶（AChE）蛋白表达的影响，以及β-细辛醚（β-asarone）的保护作用。方法：首先，将对数生长期的PC12细胞随机分为正常对照组、ACM（Aβ1-42 3.3 μmol/L）组、ACM（Aβ1-42 10 μmol/L）组、ACM（Aβ1-42 30 μmol/L）组，分别采用实时无标记动态细胞分析技术（Real Time Cell Analysis, RTCA）和MTT法检测各组PC12细胞的存活率，Western blot检测PC12细胞BDNF、AChE蛋白的表达；其次，将对数生长期的PC12细胞随机分为正常对照组、ACM（Aβ1-42 10 μmol/L）组、β-asarone（18.5、55.5、166.7 μg/mL）组，RTCA法检测PC12细胞存活率的改变，Western blot检测PC12细胞BDNF、AChE蛋白表达的变化。结果：ACM作用24 h，各组PC12细胞存活率无统计学差异（P>0.05），ACM作用36 h，ACM（Aβ1-42 10 μmol/L）组、ACM（Aβ1-42 30 μmol/L）组PC12细胞存活率显著降低(P<0.01)；BDNF、AChE蛋白表达随Aβ1-42 浓度增加显著升高(P<0.05或P<0.01)；β-asarone(18.5、55.5、166.7 μg/mL)能显著提高PC12细胞的存活率，β-asarone(55.5、166.7 μg/mL)组AChE表达相比于模型组显著下降(P<0.05或P<0.01)，β-asarone(55.5 μg/mL)组与模型组比较BDNF表达增加(P<0.05)。结论：β-asarone能抑制AChE的上调，对BDNF的表达有促进作用，提示β-asarone对神经元细胞有一定的保护作用。

关键词: β-asarone, Aβ_1-42, ACM, PC12细胞, BDNF, AChE

Abstract:

AIM: To investigate the effect of different concentrations of Aβ _1-42 activated-ACM cell on the viability and expression of BDNF, AChE of PC12 cells, and to explore the protective effects of β-asarone on the PC12 cells. METHODS: The logarithmic growth PC12 cells were randomly divided into normal group, ACM (Aβ_1-42 3.3 μmol/L) group, ACM (Aβ_1-42 10 μmol/L) group and ACM (Aβ_1-42 30 μmol/L) group. The Real time cell analysis (RTCA) system and MTT assay were used to detect the survival rate of PC12 cells, respectively; Western blot assay was used to detect the expression of BDNF, AChE. The logarithmic growth PC12 cells divided into normal group, ACM (Aβ_1-42 μmol/L) group, β-asarone(18.5 μg/mL, 55.5 μg/mL, 166.7 μg/mL) group randomly. The change of survival rate of PC12 cells was detected by RTCA assay; the expression of BDNF, AChE were detected by Western blot. RESULTS: There was no significant difference in the survival rate of PC12 cells in each group after ACM activated for 24 h (P>0.05); while the survival rate of PC12 cells was significantly reduced in ACM (Aβ_1-42 10 μmol/L) group and ACM (Aβ_1-42 30 μmol/L) group (P<0.01) after ACM activated for 36 h; the expression of BDNF, AChE protein increased significantly with the increase of the concentration of Aβ_1-42 (P<0.05 or P<0.01); the survival rate of PC12 cells were significantly increased with β-asarone (18.5 μg/mL, 55.5 μg/mL, 166.7 μg/mL); the expression of AChE in β-asarone (55.5 μg/mL, 166.7 μg/mL) group was significantly lower than that in the model group (P<0.05 or P<0.01); the expression of BDNF in β-asarone (55.5 μg/mL) group was increased compared with that of the model group (P<0.05). CONCLUSION: β-asarone can inhibit the increase of AChE expression, and promote the expression of BDNF, suggesting that β-asarone has a certain protective effect on neurons.

Key words: β-asarone, Aβ_1-42, ACM, PC12 cells, BDNF, AChE

中图分类号:

R965.2

何潆，包玉婷，姚莹，宣玲，杨元宵. β-asarone对Aβ_1-42激活的ACM所致PC12细胞损伤的保护作用研究[J]. 中国临床药理学与治疗学, 2018, 23(6): 608-613.

HE Yin, BAO Yuting, YAO Ying, XUAN Ling, YANG Yuanxiao. Protective effects of β-asarone on PC12 cells injury induced by Aβ_1-42 activated ACM[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2018, 23(6): 608-613.

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β-asarone对Aβ_1-42激活的ACM所致PC12细胞损伤的保护作用研究

Protective effects of β-asarone on PC12 cells injury induced by Aβ_1-42 activated ACM

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β-asarone对Aβ1-42激活的ACM所致PC12细胞损伤的保护作用研究

Protective effects of β-asarone on PC12 cells injury induced by Aβ1-42 activated ACM

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β-asarone对Aβ_1-42激活的ACM所致PC12细胞损伤的保护作用研究

Protective effects of β-asarone on PC12 cells injury induced by Aβ_1-42 activated ACM