PMID- 29570219 OWN - NLM STAT- MEDLINE DCOM- 20191029 LR - 20211204 IS - 1096-9896 (Electronic) IS - 0022-3417 (Print) IS - 0022-3417 (Linking) VI - 245 IP - 2 DP - 2018 Jun TI - Advanced glycation end-products suppress autophagic flux in podocytes by activating mammalian target of rapamycin and inhibiting nuclear translocation of transcription factor EB. PG - 235-248 LID - 10.1002/path.5077 [doi] AB - Insufficient autophagy in podocytes is related to podocyte injury in diabetic nephropathy (DN). Advanced glycation end-products (AGEs) are major factors of podocyte injury in DN. However, the role and mechanism of AGEs in autophagic dysfunction remain unknown. We investigated autophagic flux in AGE-stimulated cultured podocytes using multiple assays: western blotting, reverse transcription-quantitative PCR, immunofluorescence staining, and electron microscopy. We also utilized chloroquine and a fluorescent probe to monitor the formation and turnover of autophagosomes. Mice of the db/db strain were used to model diabetes mellitus (DM) with high levels of AGEs. To mimic DM with normal levels of AGEs as a control, we treated db/db mice with pyridoxamine to block AGE formation. AGEs impaired autophagic flux in the cultured podocytes. Compared with db/db mice with normal AGEs but high glucose levels, db/db mice with high AGEs and high glucose levels exhibited lower autophagic activity. Aberrant autophagic flux was related to hyperactive mammalian target of rapamycin (mTOR), a major suppressor of autophagy. Pharmacologic inhibition of mTOR activity restored impaired autophagy. AGEs inhibited the nuclear translocation and activity of the pro-autophagic transcription factor EB (TFEB) and thus suppressed transcription of its several autophagic target genes. Conversely, TFEB overexpression prevented AGE-induced autophagy insufficiency. Attenuating mTOR activity recovered TFEB nuclear translocation under AGE stimulation. Co-immunoprecipitation assays further demonstrated the interaction between mTOR and TFEB in AGE-stimulated podocytes and in glomeruli from db/db mice. In conclusion, AGEs play a crucial part in suppressing podocyte autophagy under DM conditions. AGEs inhibited the formation and turnover of autophagosomes in podocytes by activating mTOR and inhibiting the nuclear translocation of TFEB. (c) 2018 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of Pathological Society of Great Britain and Ireland. CI - (c) 2018 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of Pathological Society of Great Britain and Ireland. FAU - Zhao, Xingchen AU - Zhao X AUID- ORCID: 0000-0002-2262-0863 AD - The Second School of Clinical Medicine, Southern Medical University, Guangzhou, Guangdong, PR China. AD - Division of Nephrology, Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangdong Provincial Institute of Geriatrics, Guangzhou, Guangdong, PR China. FAU - Chen, Yuanhan AU - Chen Y AD - The Second School of Clinical Medicine, Southern Medical University, Guangzhou, Guangdong, PR China. AD - Division of Nephrology, Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangdong Provincial Institute of Geriatrics, Guangzhou, Guangdong, PR China. FAU - Tan, Xiaofan AU - Tan X AD - Division of Nephrology, Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangdong Provincial Institute of Geriatrics, Guangzhou, Guangdong, PR China. AD - Division of Nephrology, Zhongshan City People's Hospital, Zhongshan Hospital of Sun Yat-sen University, Zhongshan, Guangdong, PR China. FAU - Zhang, Li AU - Zhang L AD - Division of Nephrology, Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangdong Provincial Institute of Geriatrics, Guangzhou, Guangdong, PR China. FAU - Zhang, Hong AU - Zhang H AD - Division of Nephrology, Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangdong Provincial Institute of Geriatrics, Guangzhou, Guangdong, PR China. FAU - Li, Zhilian AU - Li Z AD - Division of Nephrology, Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangdong Provincial Institute of Geriatrics, Guangzhou, Guangdong, PR China. FAU - Liu, Shuangxin AU - Liu S AD - Division of Nephrology, Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangdong Provincial Institute of Geriatrics, Guangzhou, Guangdong, PR China. FAU - Li, Ruizhao AU - Li R AD - Division of Nephrology, Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangdong Provincial Institute of Geriatrics, Guangzhou, Guangdong, PR China. FAU - Lin, Ting AU - Lin T AD - Division of Nephrology, Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangdong Provincial Institute of Geriatrics, Guangzhou, Guangdong, PR China. FAU - Liao, Ruyi AU - Liao R AD - Division of Nephrology, Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangdong Provincial Institute of Geriatrics, Guangzhou, Guangdong, PR China. FAU - Zhang, Qianmei AU - Zhang Q AD - Division of Nephrology, Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangdong Provincial Institute of Geriatrics, Guangzhou, Guangdong, PR China. FAU - Dong, Wei AU - Dong W AD - Division of Nephrology, Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangdong Provincial Institute of Geriatrics, Guangzhou, Guangdong, PR China. FAU - Shi, Wei AU - Shi W AD - The Second School of Clinical Medicine, Southern Medical University, Guangzhou, Guangdong, PR China. AD - Division of Nephrology, Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangdong Provincial Institute of Geriatrics, Guangzhou, Guangdong, PR China. FAU - Liang, Xinling AU - Liang X AD - Division of Nephrology, Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangdong Provincial Institute of Geriatrics, Guangzhou, Guangdong, PR China. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20180430 PL - England TA - J Pathol JT - The Journal of pathology JID - 0204634 RN - 0 (Basic Helix-Loop-Helix Leucine Zipper Transcription Factors) RN - 0 (Glycation End Products, Advanced) RN - 0 (TFEB protein, human) RN - 0 (Tcfeb protein, mouse) RN - 0 (advanced glycation end products-bovine serum albumin) RN - 27432CM55Q (Serum Albumin, Bovine) RN - EC 2.7.1.1 (MTOR protein, human) RN - EC 2.7.1.1 (mTOR protein, mouse) RN - EC 2.7.11.1 (TOR Serine-Threonine Kinases) SB - IM MH - Active Transport, Cell Nucleus/drug effects MH - Adult MH - Animals MH - Autophagosomes/*drug effects/metabolism/ultrastructure MH - Autophagy/*drug effects MH - Basic Helix-Loop-Helix Leucine Zipper Transcription Factors/genetics/*metabolism MH - Cells, Cultured MH - Diabetic Nephropathies/genetics/*metabolism/pathology MH - Disease Models, Animal MH - Female MH - Glycation End Products, Advanced/*toxicity MH - Humans MH - Male MH - Mice, Inbred C57BL MH - Middle Aged MH - Podocytes/*drug effects/metabolism/ultrastructure MH - Serum Albumin, Bovine/*toxicity MH - Signal Transduction/drug effects MH - TOR Serine-Threonine Kinases/*metabolism PMC - PMC5969319 OTO - NOTNLM OT - advanced glycation end-products OT - autophagy OT - diabetic nephropathy OT - mTOR OT - podocytes OT - transcription factor EB EDAT- 2018/03/24 06:00 MHDA- 2019/10/30 06:00 PMCR- 2018/05/25 CRDT- 2018/03/24 06:00 PHST- 2017/09/07 00:00 [received] PHST- 2018/02/12 00:00 [revised] PHST- 2018/03/14 00:00 [accepted] PHST- 2018/03/24 06:00 [pubmed] PHST- 2019/10/30 06:00 [medline] PHST- 2018/03/24 06:00 [entrez] PHST- 2018/05/25 00:00 [pmc-release] AID - PATH5077 [pii] AID - 10.1002/path.5077 [doi] PST - ppublish SO - J Pathol. 2018 Jun;245(2):235-248. doi: 10.1002/path.5077. Epub 2018 Apr 30.