枸杞多糖通过泛素蛋白酶体途径降解突变亨廷顿蛋白

发布时间：2018-07-10 19:55

本文选题：突变亨廷顿蛋白 + 枸杞多糖　；参考：《中国临床解剖学杂志》2017年06期

【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。
[Abstract]:Objective to investigate the degradation pathway of mutant huntingtintin-mHtt by Lycium barbarum polysaccharide. Methods in HEK293 cells which expressed mHT160Q stably, the activity of caspase-3 was detected by different concentration of LBPnCCK8 method. The effect of LBP on mHTT in HEK293-160Q cells was detected by fluorescence microscope and RT-PCR was used to detect whether LBP affected the mRNA level of HEK293-160Q cells, and HEK293-160Q cells were treated with LBP MG132 and chloroquine, and the changes of mHTT in different cells were detected by Western blot. Results LBP could increase the activity of HEK293-160Q cells, reduce the activity of caspase-3 and reduce the mRNAs of HEK293-160Q cells. Both LBP and chloroquine had no effect on the degradation of mHTT. Conclusion LBP can degrade mHttvia ubiquitin proteasome pathway, reduce the cytotoxicity induced by mHtt, increase cell viability and inhibit apoptosis.
【作者单位】：桂林医学院人体解剖学教研室;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;
【基金】：广西高校中青年教师基础能力提升项目(2017KY0497) 广西自治区自然科学基金青年项目(2017GXNSFBA198155)
【分类号】：R742.2

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