title: Novel role for TRPC4 in regulation of macroautophagy by a small molecule in vascular endothelial cells.
abstract: Macroautophagy (autophagy) is an important factor affecting the function of vascular endothelial cells (VECs) and must be tightly regulated in these cells. However, the precise mechanisms underlying this process, particularly in the presence of serum, remain obscure. In this study, we identified trans-3,5,4'-trimethoxystilbene (TMS) as a potent small molecule inducer of autophagy in human umbilical vascular endothelial cells (HUVECs) in the presence of serum. Using high-throughput DNA microarray and siRNA transfection technologies, we demonstrated that TMS induced autophagy by up-regulating the expression of the transient receptor potential canonical channel 4 (TRPC4), an important cation channel in HUVECs. In addition, the overexpression of TRPC4 by plasmid transfection also induced autophagy. Mechanistic studies revealed that the up-regulation of TRPC4 increased the intracellular Ca²⁺ concentration, which, in turn, activated the Ca²⁺/CaMKKβ/AMPK pathway, leading to mTOR inhibition and autophagy. Our study identifies a novel role for TRPC4 in the regulation of autophagy in VECs. TMS is a useful new tool for investigating the molecular mechanism of autophagy in VECs and may serve as a potential lead compound for developing a class of autophagy inducers to treat autophagy-related diseases.
authors.affiliation: College of Bioengineering, Henan University of Technology, Lianhua Street, Zhengzhou, 450001, China. Electronic address: chaperones@163.com.
authors.forename: Lu
authors.initials: L
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Applied Organic Chemistry, Lanzhou University, 222 Tianshui Street S., Lanzhou 730000, China.
authors.forename: Fang
authors.initials: F
authors.lastname: Dai
authors.affiliation: College of Bioengineering, Henan University of Technology, Lianhua Street, Zhengzhou, 450001, China.
authors.forename: LiuQing
authors.initials: L
authors.lastname: Cui
authors.affiliation: College of Bioengineering, Henan University of Technology, Lianhua Street, Zhengzhou, 450001, China.
authors.forename: Hongjuan
authors.initials: H
authors.lastname: Jing
authors.affiliation: College of Bioengineering, Henan University of Technology, Lianhua Street, Zhengzhou, 450001, China.
authors.forename: Pei
authors.initials: P
authors.lastname: Fan
authors.affiliation: College of Bioengineering, Henan University of Technology, Lianhua Street, Zhengzhou, 450001, China.
authors.forename: Xiaorong
authors.initials: X
authors.lastname: Tan
authors.affiliation: Department of Obstetrics and Gynecology, The Third Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China.
authors.forename: YuQi
authors.initials: Y
authors.lastname: Guo
authors.affiliation: College of Bioengineering, Henan University of Technology, Lianhua Street, Zhengzhou, 450001, China.
authors.forename: GuangZhou
authors.initials: G
authors.lastname: Zhou
mesh_terms: D055372:AMP-Activated Protein Kinases / Q000378:metabolism; D001343:Autophagy / Q000187:drug effects* / Q000235:genetics; D002118:Calcium / Q000378:metabolism; D054737:Calcium-Calmodulin-Dependent Protein Kinase Kinase / Q000378:metabolism; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D042541:Intracellular Space / Q000187:drug effects / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D013267:Stilbenes / Q000494:pharmacology*; D058570:TOR Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D050052:TRPC Cation Channels / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D015854:Up-Regulation / Q000187:drug effects / Q000235:genetics
chemical_list: C497286:3,4',5-trimethoxystilbene; D013267:Stilbenes; D050052:TRPC Cation Channels; C489571:TRPC4 ion channel; C546842:MTOR protein, human; D058570:TOR Serine-Threonine Kinases; D054737:Calcium-Calmodulin-Dependent Protein Kinase Kinase; D055372:AMP-Activated Protein Kinases; C526180:PRKAA1 protein, human; D002118:Calcium
keywords: Macroautophagy; Mammalian target of rapamycin; Trans-3,5,4′-trimethoxystilbene; Transient receptor potential canonical channel 4; Vascular endothelial cell