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《China Journal of Chinese Materia Medica》 2015-10
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Molecular mechanism of emodin on inhibiting autophagy induced by HBSS in renal tubular cells

HU Hao;SUN Wei;GU Liu-bao;TU Yue;LIU Hong;Research Institute of Kidney Disease,Nanjing University of Chinese Medicine;Department of Nephrology,Jiangsu Provincial Hospital of Chinese Medicine,The Affiliated Hospital of Nanjing University of Chinese Medicine;Department of Endocrinology,Jiangsu Province Official Hospital;  
Objective: To explore the regulative effects and possible mechanisms of emodin on autophagy induced by starvation in rat' s renal tubular epithelial cells( NRK-52E). Method: Firstly,Hank's balanced salt solution( HBSS) was used to induce starvation and the protein expression of microtubule-associated protein 1 light chain 3( LC3) Ⅰ / Ⅱ,an autophagic marker of mammalian congener,was detected by Western blot with or without the treatment of emodin. Secondly,the changes of redfluorescent protein-microtubule associated protein light chain3( RFP-LC3) fluorescent particles,treated by HBSS( 1 m L) and bafilomycin A1( 10 nmol·L-1) with or without emodin,were observed through fluorescence microscopy in NRK-52 E cells transient transfected by RFP-LC3 plasmid. With the intervention of mammalian target of rapamycin m TOR inhibitor rapamycin( 100 nmol·L-1),the effect of blocking m TOR signaling pathway on autophagic inhibition of emodin was observed. Finally,the effect of m TOR signaling pathway on autophagic inhibition of emodin was further evaluated through the over-expression of endogenous m TOR inhibitory protein DEP domain-containing m TOR-interacting protein( DEPTOR). Result: HBSS hunger could induce high protein expression of LC3Ⅱ in NRK-52 E cells,and the intervention of emodin could reverse the unregulated protein expression of LC3Ⅱ induced by HBSS. The number of RFP-LC3 fluorescent particles was increased after the co-treatment of HBSS and bafilomycin A1,and this increase was inhibited by emodin. After the co-treatment of rapamycin,emodin and HBSS,the LC3Ⅱ protein expression restored in NRK-52 E cells,compared with the treatment of HBSS. Over-expression of DEPTOR could also block the inhibitive effect of emodin on LC3 Ⅱ protein expression. Conclusion: Emodin could inhibit HBSS-induced LC3Ⅱ protein expression and the activation of autophagy in NRK-52 E cells,and the effect of blocking autophagy may be mediated through m TOR signaling pathway.
【Fund】: 国家自然科学基金面上项目(81373607);; 江苏高校优势学科建设工程资助项目
【CateGory Index】: R285.5
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1 Xu Yiling;Wang Guodong;Liu Bo;Yu Bo;Huang Qifei;Su Xiuyuan;Liu Huixia;Department of Geriatrics,Xiangya Hospital of Central South University;;Emodin reduces FFAs-induced fatty degeneration in HepG2 cells via the AMPK/SREBP-1 pathway[J];中国医师杂志;2017-04
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