Volume 4, Issue 3, May 2018, Page: 51-56
Cannabidiol Attenuates Palmitic Acid-Induced Injury in Cultured Hepatocytes Through Promoting Autophagic Flux
Rui Chen, Organ Transplantation Center, Changhai Hospital of Naval Medical University, Shanghai, China
Xiaogang Gao, Organ Transplantation Center, Changhai Hospital of Naval Medical University, Shanghai, China
Lei Zhang, Organ Transplantation Center, Changhai Hospital of Naval Medical University, Shanghai, China
Wenyu Zhao, Organ Transplantation Center, Changhai Hospital of Naval Medical University, Shanghai, China
Li Zeng, Organ Transplantation Center, Changhai Hospital of Naval Medical University, Shanghai, China
Youhua Zhu, Organ Transplantation Center, Changhai Hospital of Naval Medical University, Shanghai, China
Zhiren Fu, Organ Transplantation Center, Changzheng Hospital of Naval Medical University, Shanghai, China
Received: Jun. 21, 2018;       Accepted: Aug. 2, 2018;       Published: Aug. 31, 2018
DOI: 10.11648/j.ijcems.20180403.15      View  506      Downloads  30
Abstract
Objective: This work was designed to investigate the protection of cannabidiol (CBD) against palmitic acid (PA)-induced injury in cultured hepatocytes and the underlying mechanism associated with autophagic flux. Methods: Experiment 1: Primary cultured hepatocytes were stimulated with PA (800 μmol/L) and treated with CBD (5 μmol/L) and chloroquine (CQ, 50 nmol/L) or not for 24 hours (1: control group; 2: PA-stimulated group; 3: PA-stimulated group treated with CBD; 4: PA-stimulated group treated with CBD and CQ). Autophagic flux was evaluated by Western blot analysis. Apoptosis was measured by flow cytometry. The mRNA expression of genes involved in endoplasmic reticulum stress was determined by reverse transcription PCR. The mitochondrial function was determined by using fluorescent probe including Rh123 and lucigenin. Experiment 2: Primary cultured hepatocytes were treated with CBD alone for 24 h (1: control group; 2: lower-dose CBD-treated group; 3: higher-dose CBD-treated group). Then, the autophagic flux was evaluated by Western blot analysis. Results: When compared to control group, exposure to PA significantly led to impaired autopagic flux (evidenced by increased ratio of LC3-II/LC3-I and protein expression of p62), increased apoptosis, endoplasmic reticulum stress (evidenced by increased mRNA expression of C/EBP homologous protein, glucose-regulated protein 78, and X-box protein 1), and mitochondrial dysfunction (evidenced by reduced mitochondrial membrane potential and enhanced formation of mitochondrial reactive oxygen species). When compared to PA-stimulated group, CBD treatment significantly attenuated PA-induced impaired autophagic flux, apoptosis, endoplasmic reticulum stress, and mitochondrial dysfunction in cultured hepatocytes. The protection of CBD against PA was abolished by co-incubation with CQ. In addition, treatment with CBD alone had no significant effect on autophagic flux in cultured hepatocytes Conclusion: Cannabidiol attenuates palmitic acid-induced impaired autophagic flux, apoptosis, endoplasmic reticulum stress, and mitochondrial dysfunction in cultured hepatocytes through promoting autophagic flux.
Keywords
Cannabidiol, Palmitic Acid, Autophagic Flux, Hepatocytes, Apoptosis, Mitochondrial Dysfunction, Endoplasmic Reticulum Stress
To cite this article
Rui Chen, Xiaogang Gao, Lei Zhang, Wenyu Zhao, Li Zeng, Youhua Zhu, Zhiren Fu, Cannabidiol Attenuates Palmitic Acid-Induced Injury in Cultured Hepatocytes Through Promoting Autophagic Flux, International Journal of Clinical and Experimental Medical Sciences. Vol. 4, No. 3, 2018, pp. 51-56. doi: 10.11648/j.ijcems.20180403.15
Copyright
Copyright © 2018 Authors retain the copyright of this article.
This article is an open access article distributed under the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Reference
[1]
HUANG X, XU M, CHEN Y, PENG K, HUANG Y, WANG P, et al. Validation of the fatty liver index for nonalcoholic fatty liver disease in middle-aged and elderly Chinese [J]. Medicine (Baltimore), 2015, 94: e1682. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4616754/.
[2]
JACOBS DS, KOHUT SJ, JIANG S, NIKAS SP, MAKRIYANNIS A, BERGMAN J. Acute and chronic effects of cannabidiol on Δ⁹-tetrahydrocannabinol (Δ⁹-THC)-induced disruption in stop signal task performance [J]. Exp Clin Psychopharmacol, 2016, 24:320-330. http://content.apa.org/journals/pha/24/5/320.
[3]
PISANTI S, MALFITANO AM, CIAGLIA E, LAMBERTI A, RANIERI R, CUOMO G, et al. Cannabidiol: State of the art and new challenges for therapeutic applications [J]. Pharmacol Ther, 2017, (in press). http://www.sciencedirect.com/science/article/pii/S0163725817300657.
[4]
SILVESTRI C, PARIS D, MARTELLA A, MELCK D, GUADAGNINO I, CAWTHORNE M, et al. Two non-psychoactive cannabinoids reduce intracellular lipid levels and inhibit hepatosteatosis [J]. J Hepatol, 2015, 62:1382-1390. https://linkinghub.elsevier.com/retrieve/pii/S0168-8278 (15) 00003-3.
[5]
BARREYRO FJ, KOBAYASHI S, BRONK SF, WERNEBURG NW, MALHI H, GORES GJ. Transcriptional regulation of Bim by FoxO3A mediates hepatocyte lipoapoptosis [J]. J Biol Chem, 2007, 282:27141-27154. http://www.jbc.org/cgi/pmidlookup?view=long&pmid=17626006.
[6]
MALHI H, BRONK SF, WERNEBURG NW, GORES GJ. Free fatty acids induce JNK-dependent hepatocyte lipoapoptosis [J]. J Biol Chem, 2006, 281:12093-12101. http://www.jbc.org/cgi/pmidlookup?view=long&pmid=16505490.
[7]
GONZÁLEZ-RODRÍGUEZ A, MAYORAL R, AGRA N, VALDECANTOS MP, PARDO V, MIQUILENA-COLINA ME, et al. Impaired autophagic flux is associated with increased endoplasmic reticulum stress during the development of NAFLD [J]. Cell Death Dis, 2014, 5:e1179. http://dx.doi.org/10.1038/cddis.2014.162.
[8]
LIU A, HUANG L, GUO E, LI R, YANG J, LI A, et al. Baicalein pretreatment reduces liver ischemia/reperfusion injury via induction of autophagy in rats [J]. Sci Rep, 2016, 6:25042. http://dx.doi.org/10.1038/srep25042.
[9]
ANDING AL, BAEHRECKE EH. Autophagy in cell life and cell death [J]. Curr Top Dev Biol, 2015, 114:67-91. https://linkinghub.elsevier.com/retrieve/pii/S0070-2153 (15) 00048-4.
[10]
ZHANG XJ, CHEN S, HUANG KX, LE WD. Why should autophagic flux be assessed [J]? Acta Pharmacol Sin, 2013, 34:595-599. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4002868/.
[11]
OUYANG L, SHI Z, ZHAO S, WANG FT, ZHOU TT, LIU B, BAO JK. Programmed cell death pathways in cancer: a review of apoptosis, autophagy and programmed necrosis [J]. Cell Prolif, 2012, 45:487-498. http://onlinelibrary.wiley.com/doi/10.1111/j.1365-2184.2012.00845.x/abstract;jsessionid=6A05E1100427B10093982C7765C242CF.f03t04.
[12]
CASAREJOS MJ, PERUCHO J, GOMEZ A, MUÑOZ MP, FERNANDEZ-ESTEVEZ M, SAGREDO O, et al. Natural cannabinoids improve dopamine neurotransmission and tau and amyloid pathology in a mouse model of tauopathy [J]. J Alzheimers Dis, 2013, 35:525-539. http://content.iospress.com/openurl?genre=article&id=doi:10.3233/JAD-130050.
[13]
HOSSEINZADEH M, NIKSERESHT S, KHODAGHOLI F, NADERI N, MAGHSOUDI N. Cannabidiol post-treatment alleviates rat epileptic-related behaviors and activates hippocampal cell autophagy pathway along with antioxidant defense in chronic phase of pilocarpine-induced seizure [J]. J Mol Neurosci, 2016, 58:432-440. https://dx.doi.org/10.1007/s12031-015-0703-6.
[14]
YANG L, ROZENFELD R, WU D, DEVI LA, ZHANG Z, CEDERBAUM A. Cannabidiol protects liver from binge alcohol-induced steatosis by mechanisms including inhibition of oxidative stress and increase in autophagy [J]. Free Radic Biol Med, 2014, 68:260–267. https://linkinghub.elsevier.com/retrieve/pii/S0891-5849 (13) 01567-0.
[15]
CHEN R, WANG Q, SONG S, LIU F, HE B, GAO X. Protective role of autophagy in methionine-choline deficient diet-induced advanced nonalcoholic steatohepatitis in mice [J]. Eur J Pharmacol, 2016, 770:126-133. https://linkinghub.elsevier.com/retrieve/pii/S0014-2999 (15) 30350-2.
[16]
BOZAYKUT P, SAHIN A, KARADEMIR B, OZER NK. Endoplasmic reticulum stress related molecular mechanisms in nonalcoholic steatohepatitis [J]. Mech Ageing Dev, 2016, 157:17-29. https://linkinghub.elsevier.com/retrieve/pii/S0047-6374 (16) 30102-6.
[17]
CABRÉ N, CAMPS J, JOVEN J. Inflammation, mitochondrial metabolism and nutrition: the multi-faceted progression of non-alcoholic fatty liver disease to hepatocellular carcinoma [J]. Hepatobiliary Surg Nutr, 2016, 5:438-443. http://dx.doi.org/10.21037/hbsn.2016.09.11.
[18]
LIU K, ZHAO Q, LIU P, CAO J, GONG J, WANG C, et al. ATG3-dependent autophagy mediates mitochondrial homeostasis in pluripotency acquirement and maintenance [J]. Autophagy, 2016, 12:2000-2008. http://www.tandfonline.com/doi/full/10.1080/15548627.2016.1212786.
[19]
WU D, CEDERBAUM AI. Inhibition of autophagy promotes CYP2E1-dependent toxicity in HepG2 cells via elevated oxidative stress, mitochondria dysfunction and activation of p38 and JNK MAPK [J]. Redox Biol, 2013, 1:552-565. https://linkinghub.elsevier.com/retrieve/pii/S2213-2317 (13) 00077-3.
[20]
CHEN R, GAO XG, ZHANG L, SHI XM, GUO WY, ZHU YH, et al. Therapeutic effect and mechanism of cannabidiol on nonalcoholic fatty liver disease in rats [J]. Med J Chin PLA, 2017, 42:515-519.
Browse journals by subject