title: Cinnamaldehyde ameliorates LPS-induced cardiac dysfunction via TLR4-NOX4 pathway: The regulation of autophagy and ROS production.
abstract: Cinnamaldehyde (CA), a major bioactive compound extracted from the essential oil of Cortex Cinnamomi, exhibits anti-inflammatory activity on endotoxemia. Accumulating evidence indicates reactive oxygen species (ROS) and autophagy play a vital role in the cardiac dysfunction during endotoxemia. The aim of this study was to unveil the mechanism of CA on ROS production and autophagy during endotoxemia. Male Sprague-Dawley rats were stimulated by LPS (20mg/kg i.v.) with or without treatment of CA. Cardiac function and histopathological staining were preformed 4h after LPS stimulation. The levels of TNF-α, IL-1β and IL-6 were detected by ELISA. The expression of p-JNK, p-ERK, p-p38, TLR4, NOX4, NOX2, ATG5 and LC3 proteins were determined by Western blot. The results showed that CA inhibited cardiac dysfunction, inflammatory infiltration and the levels of TNF-α, IL-1β and IL-6 in LPS stimulated rats by blocking the TLR4, NOX4, MAPK and autophagy signalings. In order to obtain further confirmation of the mechanism of CA on endotoxemia in vitro, a limited time-course study was firstly performed by Western blot. TLR4, NOX4 and LC3 were significantly increased after 4h LPS stimulation. CA reversed the intracellular ROS production and MAPK signaling activation induced by LPS. Electron microscopy, mRFP-GFP-LC3 transfection and western blot results revealed autophagic flux were attenuated after CA treatment. The siRNA and molecular docking results suggest that CA can suppress both TLR4 and NOX4 during endotoxemia. Our data revealed that CA ameliorated LPS-induced cardiac dysfunction by inhibiting ROS production and autophagy through TLR4-NOX4 pathway.
authors.affiliation: Institute of Materia Medica, School of Pharmacy, Fourth Military Medical University, Xi'an 710032, China; The Cultivation Project of Collaborative Innovation Center for Chinese Medicine in QinBa Mountains, Xi'an 710032, China.
authors.forename: Hang
authors.initials: H
authors.lastname: Zhao
authors.affiliation: Institute of Materia Medica, School of Pharmacy, Fourth Military Medical University, Xi'an 710032, China; The Cultivation Project of Collaborative Innovation Center for Chinese Medicine in QinBa Mountains, Xi'an 710032, China.
authors.forename: Meng
authors.initials: M
authors.lastname: Zhang
authors.affiliation: Department of Obstetrics and Gynecology, Xijing Hospital, The First Affiliated Hospital, Fourth Military Medical University, Xi'an 710032, China.
authors.forename: Fuxing
authors.initials: F
authors.lastname: Zhou
authors.affiliation: Institute of Materia Medica, School of Pharmacy, Fourth Military Medical University, Xi'an 710032, China; The Cultivation Project of Collaborative Innovation Center for Chinese Medicine in QinBa Mountains, Xi'an 710032, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Cao
authors.affiliation: Institute of Materia Medica, School of Pharmacy, Fourth Military Medical University, Xi'an 710032, China; The Cultivation Project of Collaborative Innovation Center for Chinese Medicine in QinBa Mountains, Xi'an 710032, China.
authors.forename: Linlin
authors.initials: L
authors.lastname: Bi
authors.affiliation: Institute of Materia Medica, School of Pharmacy, Fourth Military Medical University, Xi'an 710032, China; The Cultivation Project of Collaborative Innovation Center for Chinese Medicine in QinBa Mountains, Xi'an 710032, China.
authors.forename: Yanhua
authors.initials: Y
authors.lastname: Xie
authors.affiliation: Institute of Materia Medica, School of Pharmacy, Fourth Military Medical University, Xi'an 710032, China; The Cultivation Project of Collaborative Innovation Center for Chinese Medicine in QinBa Mountains, Xi'an 710032, China. Electronic address: yangqian@fmmu.edu.cn.
authors.forename: Qian
authors.initials: Q
authors.lastname: Yang
authors.affiliation: Institute of Materia Medica, School of Pharmacy, Fourth Military Medical University, Xi'an 710032, China; The Cultivation Project of Collaborative Innovation Center for Chinese Medicine in QinBa Mountains, Xi'an 710032, China. Electronic address: wangsiw@fmmu.edu.cn.
authors.forename: Siwang
authors.initials: S
authors.lastname: Wang
mesh_terms: D000171:Acrolein / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology; D000818:Animals; D001343:Autophagy / Q000187:drug effects; D015415:Biomarkers; D016207:Cytokines / Q000096:biosynthesis; D004195:Disease Models, Animal; D004452:Echocardiography; D018836:Inflammation Mediators; D008070:Lipopolysaccharides / Q000009:adverse effects; D020935:MAP Kinase Signaling System / Q000187:drug effects; D008297:Male; D032383:Myocytes, Cardiac; D000074663:NADPH Oxidase 4; D019255:NADPH Oxidases / Q000235:genetics / Q000378:metabolism*; D034622:RNA Interference; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects*; D051197:Toll-Like Receptor 4 / Q000235:genetics / Q000378:metabolism*; D018754:Ventricular Dysfunction / Q000175:diagnosis / Q000188:drug therapy / Q000209:etiology* / Q000378:metabolism*
chemical_list: D015415:Biomarkers; D016207:Cytokines; D018836:Inflammation Mediators; D008070:Lipopolysaccharides; D017382:Reactive Oxygen Species; D051197:Toll-Like Receptor 4; D000171:Acrolein; D000074663:NADPH Oxidase 4; D019255:NADPH Oxidases; C498911:Nox4 protein, rat; C012843:cinnamaldehyde
keywords: Autophagy; Cinnamaldehyde; Endotoxemia; NADPH oxidase; Toll like receptor