Palmitic acid increases apoptosis by mitochondrial pathway in hepatocytes with growth hormone deficiency

Objective To investigate the specific molecular mechanisms of palmitic acid (PA)-induced lipid deposition in cultured hepatocytes with knockdown of growth hormone receptor (GHR). Methods Hepatic L02 cells were treated with different concentrations of PA (0, 75, 150 and 300 μmol/L) for 12, 24, and 48...

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Autores principales:	WANG Yunting, ZHENG Xiaoya, CHEN Yue, QIAN Wenjie, WU Xun, REN Wei
Formato:	article
Lenguaje:	ZH
Publicado:	Editorial Office of Journal of Third Military Medical University 2021
Materias:	growth hormone receptor knockdown mitochondria reactive oxygen species apoptosis Medicine (General) R5-920
Acceso en línea:	https://doaj.org/article/61b303d66db0433485686ccce54028f5
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spelling	oai:doaj.org-article:61b303d66db0433485686ccce54028f52021-11-12T05:02:59ZPalmitic acid increases apoptosis by mitochondrial pathway in hepatocytes with growth hormone deficiency10.16016/j.1000-5404.2021041501000-5404https://doaj.org/article/61b303d66db0433485686ccce54028f52021-11-01T00:00:00Zhttp://aammt.tmmu.edu.cn/Upload/rhtml/202104150.htmhttps://doaj.org/toc/1000-5404Objective To investigate the specific molecular mechanisms of palmitic acid (PA)-induced lipid deposition in cultured hepatocytes with knockdown of growth hormone receptor (GHR). Methods Hepatic L02 cells were treated with different concentrations of PA (0, 75, 150 and 300 μmol/L) for 12, 24, and 48 h, respectively, and cell viability was measured. Then the L02 cells were transinfected with GHR-lentiviral vector (shRNAGHR group) or control vector (vector group). Oil red "O" staining and boron-dipyrromethene (BODIPY) fluorescence staining were used to observe lipid deposition. Flow cytometry was employed to detect the apoptosis and production of reactive oxygen species (ROS). JC-1 mitochondrial membrane potential assay was applied to detect the changes in membrane potential. MitoSox fluorescence was used to detect mitochondrial ROS level. Western blotting was carried out to measure mitochondrial-mediated apoptotic protein expression. After pretreated with targeted mitochondrial antioxidant, mitoTEMPO, the cells were further observed whether it could offset PA-induced changes in ROS level and apoptosis. Results PA inhibited the survival rate of the L02 cells in a concentration-dependent manner. Compared with the control group, treatment of 150 μumol/L PA for 12, 24, and 48 h resulted in significantly increased intracellular ROS level, mitochondrial ROS level and cell apoptosis in the shRNAGHR. Oil red "O" staining and BODIPY fluorescence confirmed that shRNAGHR group was more prone to lipid deposition with same dose of PA stimulation. Western blotting showed the levels of cytochrome C protein, Cleaved-Caspase 9, Cleaved-Caspase 3, and polymerase (PARP) were significantly higher in the shRNAGHR group than the vector group (P < 0.05). Pretreatment of targeted mitochondrial antioxidant mitoTEMPO significantly ameliorated shRNAGHR induced cells apoptosis. Conclusion Knockdown of GHR aggravates lipid deposition and apoptosis in hepatocytes, and PA plays an important role in the process through mitochondria/ROS pathway.WANG YuntingZHENG XiaoyaCHEN YueQIAN WenjieWU XunREN WeiEditorial Office of Journal of Third Military Medical Universityarticlegrowth hormone receptor knockdownmitochondriareactive oxygen speciesapoptosisMedicine (General)R5-920ZHDi-san junyi daxue xuebao, Vol 43, Iss 21, Pp 2366-2374 (2021)
institution	DOAJ
collection	DOAJ
language	ZH
topic	growth hormone receptor knockdown mitochondria reactive oxygen species apoptosis Medicine (General) R5-920
spellingShingle	growth hormone receptor knockdown mitochondria reactive oxygen species apoptosis Medicine (General) R5-920 WANG Yunting ZHENG Xiaoya CHEN Yue QIAN Wenjie WU Xun REN Wei Palmitic acid increases apoptosis by mitochondrial pathway in hepatocytes with growth hormone deficiency
description	Objective To investigate the specific molecular mechanisms of palmitic acid (PA)-induced lipid deposition in cultured hepatocytes with knockdown of growth hormone receptor (GHR). Methods Hepatic L02 cells were treated with different concentrations of PA (0, 75, 150 and 300 μmol/L) for 12, 24, and 48 h, respectively, and cell viability was measured. Then the L02 cells were transinfected with GHR-lentiviral vector (shRNAGHR group) or control vector (vector group). Oil red "O" staining and boron-dipyrromethene (BODIPY) fluorescence staining were used to observe lipid deposition. Flow cytometry was employed to detect the apoptosis and production of reactive oxygen species (ROS). JC-1 mitochondrial membrane potential assay was applied to detect the changes in membrane potential. MitoSox fluorescence was used to detect mitochondrial ROS level. Western blotting was carried out to measure mitochondrial-mediated apoptotic protein expression. After pretreated with targeted mitochondrial antioxidant, mitoTEMPO, the cells were further observed whether it could offset PA-induced changes in ROS level and apoptosis. Results PA inhibited the survival rate of the L02 cells in a concentration-dependent manner. Compared with the control group, treatment of 150 μumol/L PA for 12, 24, and 48 h resulted in significantly increased intracellular ROS level, mitochondrial ROS level and cell apoptosis in the shRNAGHR. Oil red "O" staining and BODIPY fluorescence confirmed that shRNAGHR group was more prone to lipid deposition with same dose of PA stimulation. Western blotting showed the levels of cytochrome C protein, Cleaved-Caspase 9, Cleaved-Caspase 3, and polymerase (PARP) were significantly higher in the shRNAGHR group than the vector group (P < 0.05). Pretreatment of targeted mitochondrial antioxidant mitoTEMPO significantly ameliorated shRNAGHR induced cells apoptosis. Conclusion Knockdown of GHR aggravates lipid deposition and apoptosis in hepatocytes, and PA plays an important role in the process through mitochondria/ROS pathway.
format	article
author	WANG Yunting ZHENG Xiaoya CHEN Yue QIAN Wenjie WU Xun REN Wei
author_facet	WANG Yunting ZHENG Xiaoya CHEN Yue QIAN Wenjie WU Xun REN Wei
author_sort	WANG Yunting
title	Palmitic acid increases apoptosis by mitochondrial pathway in hepatocytes with growth hormone deficiency
title_short	Palmitic acid increases apoptosis by mitochondrial pathway in hepatocytes with growth hormone deficiency
title_full	Palmitic acid increases apoptosis by mitochondrial pathway in hepatocytes with growth hormone deficiency
title_fullStr	Palmitic acid increases apoptosis by mitochondrial pathway in hepatocytes with growth hormone deficiency
title_full_unstemmed	Palmitic acid increases apoptosis by mitochondrial pathway in hepatocytes with growth hormone deficiency
title_sort	palmitic acid increases apoptosis by mitochondrial pathway in hepatocytes with growth hormone deficiency
publisher	Editorial Office of Journal of Third Military Medical University
publishDate	2021
url	https://doaj.org/article/61b303d66db0433485686ccce54028f5
work_keys_str_mv	AT wangyunting palmiticacidincreasesapoptosisbymitochondrialpathwayinhepatocyteswithgrowthhormonedeficiency AT zhengxiaoya palmiticacidincreasesapoptosisbymitochondrialpathwayinhepatocyteswithgrowthhormonedeficiency AT chenyue palmiticacidincreasesapoptosisbymitochondrialpathwayinhepatocyteswithgrowthhormonedeficiency AT qianwenjie palmiticacidincreasesapoptosisbymitochondrialpathwayinhepatocyteswithgrowthhormonedeficiency AT wuxun palmiticacidincreasesapoptosisbymitochondrialpathwayinhepatocyteswithgrowthhormonedeficiency AT renwei palmiticacidincreasesapoptosisbymitochondrialpathwayinhepatocyteswithgrowthhormonedeficiency
_version_	1718431172876304384

Palmitic acid increases apoptosis by mitochondrial pathway in hepatocytes with growth hormone deficiency

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