枸杞多糖通过泛素蛋白酶体途径降解突变亨廷顿蛋白

doi:10.13418/j.issn.1001-165x.2017.06.010

中国临床解剖学杂志 ›› 2017, Vol. 35 ›› Issue (6): 641-644.doi: 10.13418/j.issn.1001-165x.2017.06.010

枸杞多糖通过泛素蛋白酶体途径降解突变亨廷顿蛋白

方方^1,2，　陈恬¹，　彭云滔¹，　李和²

1.桂林医学院人体解剖学教研室，广西桂林 541004；　2.华中科技大学同济医学院
人体解剖学系组织胚胎学教研室，武汉 430030

收稿日期:2017-07-26 出版日期:2017-11-25 发布日期:2017-12-30
通讯作者: 李和，教授，E-mail: heli_tjmu@163.com
作者简介:方方（1984-），女，广西桂林人，讲师，医学博士，主要从事神经疾病的细胞及分子生物学方面的研究，Tel：18577384365, E-mail: fangdouble@163.com
基金资助:
广西高校中青年教师基础能力提升项目（2017KY 0497）；广西自治区自然科学基金青年项目（2017GXNSFBA198155）

Lycium barbarum polysaccharide degrades the mutant huntingtin by ubiquitin-proteasome pathway

FANG Fang ^{1, 2}, CHEN Tian¹, PENG Yun-tao ¹, LI He²

1. Department of Human Anatomy, Guilin Medical University, Guilin 541004, China; 2. Division of Histology and Embryology, Department of Anatomy, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China

Received:2017-07-26 Online:2017-11-25 Published:2017-12-30

摘要/Abstract

摘要：

目的　探讨枸杞多糖（Lycium barbarum polysaccharide，LBP）降解突变亨廷顿蛋白（mutant huntingtin，mHtt）的途径。方法　在稳定表达mHtt 160Q的HEK293细胞中使用不同浓度LBP，CCK8法检测细胞活力，caspase-3活性酶标法检测caspase-3活性；使用荧光显微镜检测、Image Pro Plus 6.0分析LBP对HEK293-160Q细胞中mHtt的影响并同时使用RT-PCR法检测LBP是否影响其mRNA水平；使用LBP、MG132及氯喹，分组处理HEK293-160Q细胞，通过Western Blot法检测不同组细胞中mHtt的变化。结果　LBP能提高HEK293-160Q细胞活力，降低caspase-3活性；LBP能减少细胞中mHtt且不改变其mRNA；不同药物处理HEK293-160Q细胞后，发现与只使用LBP相比，同时使用LBP与MG132会显著降低mHtt的降解，而同时使用LBP与氯喹则对mHtt的降解没有影响。结论　LBP能通过泛素蛋白酶体途径降解mHtt，减轻mHtt所引起的细胞毒性继而提高细胞活力、抑制细胞凋亡。

关键词: 突变亨廷顿蛋白, 　枸杞多糖, 　泛素蛋白酶体途径, 　自噬溶酶体途径

Abstract:

Objective　To detect the degradation pathway of the mutant huntingtin (mHtt) after treating Lycium barbarum polysaccharide (LBP).　Methods　HEK293 cells that stably expressed mHtt containing 160 glutamine repeats was treated with LBP of various doses, CCK8 assay was used to detect the cell viability, and caspase-3 activity assay kit was used to detect the caspase-3 activity; the mHtt in HEK293-160Q cells was observed under fluorescence microscope after treatment with LBP and the images were analyzed by Image Pro Plus 6.0, at the same time, to detect its mRNA; LBP, MG132 and chloroquine were used to treat HEK293-160Q cells separately, and the changes of the mHtt were determined by Western Blot. Results　LBP could increase the viability and decrease the caspase-3 activity in HEK293-160Q cells; LBP could reduce the mHtt and did not change its mRNA levels; after treating HEK293-160Q cells with the different medicines , the Western Blot results showed that compared to cells receiving only LBP treatment, the degeneration of mHtt was significantly reduced after treating LBP and MG132. No influence on mHtt after treating LBP and chloroquine was observed.　Conclusions　LBP can degrade the mHtt by ubiquitin-proteasome pathway, increase the cell viability and decrease the apoptosis by alleviating the cytotoxicity of mHtt.

Key words: Mutant huntingtin;　Lycium barbarum polysaccharide;　Ubiquitin-proteasome pathway; , Autophagy-lysosome pathway

方方,陈恬,彭云滔,李和. 枸杞多糖通过泛素蛋白酶体途径降解突变亨廷顿蛋白[J]. 中国临床解剖学杂志, 2017, 35(6): 641-644.

FANG Fang, CHEN Tian, PENG Yun-tao, LI He. Lycium barbarum polysaccharide degrades the mutant huntingtin by ubiquitin-proteasome pathway[J]. Chinese Journal Of Clinical Anatomy, 2017, 35(6): 641-644.

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枸杞多糖通过泛素蛋白酶体途径降解突变亨廷顿蛋白

Lycium barbarum polysaccharide degrades the mutant huntingtin by ubiquitin-proteasome pathway

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