Inhibition of Frizzled-2 by small interfering RNA protects rat hepatic BRL-3A cells against cytotoxicity and apoptosis induced by Hypoxia/Reoxygenation
Gastroenterol Hepatol. 2020 Mar;43(3):107-116.
doi: 10.1016/j.gastrohep.2019.02.006.
Epub 2020 Jan 18.
[Article in
English,
Spanish]
Affiliations
- 1 Department of Hepatobiliary Surgery II, State Key Laboratory of Organ Failure Research, Guangdong Provincial Research Center for Artificial Organ and Tissue Engineering, Guangzhou Clinical Research and Transformation Center for Artificial Liver, Institute of Regenerative Medicine, Zhujiang Hospital, Southern Medical University, Guangzhou 510280, Guangdong Province, China; General Surgery Department, The Second Hospital of Shenzhen Baoan People's Hospital Group, Shenzhen 518108, Guangdong Province, China.
- 2 Department of Hepatobiliary Surgery II, State Key Laboratory of Organ Failure Research, Guangdong Provincial Research Center for Artificial Organ and Tissue Engineering, Guangzhou Clinical Research and Transformation Center for Artificial Liver, Institute of Regenerative Medicine, Zhujiang Hospital, Southern Medical University, Guangzhou 510280, Guangdong Province, China.
- 3 Department of Hepatobiliary Surgery II, State Key Laboratory of Organ Failure Research, Guangdong Provincial Research Center for Artificial Organ and Tissue Engineering, Guangzhou Clinical Research and Transformation Center for Artificial Liver, Institute of Regenerative Medicine, Zhujiang Hospital, Southern Medical University, Guangzhou 510280, Guangdong Province, China. Electronic address: drgaoy@126.com.
Abstract
Frizzled-2 plays an important role in maintaining normal hepatic cell functionality. This study aimed to investigate the role of inhibition of Frizzled-2 in protecting rat liver BRL-3A cells from Hypoxia/Reoxygenation (H/R). In vitro H/R hepatic cell model was established by culturing BRL-3A cells under H/R condition. Frizzled-2 siRNA was transfected into BRL-3A cells to inhibit Frizzled-2 signaling. Wnt5a and Frizzled-2 were significantly increased in BRL-3A cells upon H/R treatment. H/R treatment induced cell cytotoxicity, the early apoptosis rate and the intracellular Ca2+ level in BRL-3A cells while silencing frizzled-2 gene decreased the H/R induced cell cytotoxicity, apoptosis and intracellular Ca2+ level. In vivo mice study further showed the up-regulation of Frizzled-2/Wnt 5 pathway and cleaved Caspase-3 expression in liver tissues under ischemia and reperfusion injury (IRI). In summary, inhibition of Frizzled-2 by its siRNA may protects BRL-3A cells by attenuating the H/R induced cell cytotoxicity and apoptosis.
Keywords:
ARNip; Apoptosis; Ca(2+) intracelular; Estudio de ratones in vivo; Frizzled-2; Hepatic BRL-3A cells; Hepatocitos BRL-3A; Hipoxia/reoxigenación; Hypoxia/Reoxygenation (H/R); In vivo mice study; Intracellular Ca(2+); siRNA.
Copyright © 2019. Publicado por Elsevier España, S.L.U.
MeSH terms
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Animals
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Apoptosis / drug effects
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Calcium Signaling / drug effects
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Caspase 3 / biosynthesis
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Caspase 3 / genetics
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Cell Hypoxia / drug effects*
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Cell Hypoxia / genetics
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Cell Line
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Frizzled Receptors / biosynthesis
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Frizzled Receptors / genetics
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Gene Expression Regulation
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Hepatocytes / drug effects*
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Hepatocytes / metabolism
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Male
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Mice
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Mice, Inbred C57BL
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RNA Interference*
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RNA, Small Interfering / genetics
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RNA, Small Interfering / pharmacology*
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Rats
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Reperfusion Injury / genetics
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Reperfusion Injury / metabolism
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Reperfusion Injury / prevention & control*
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Wnt Signaling Pathway / drug effects
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Wnt-5a Protein / biosynthesis
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Wnt-5a Protein / genetics
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beta Catenin / biosynthesis
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beta Catenin / genetics
Substances
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CTNNB1 protein, mouse
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Frizzled Receptors
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Fzd2 protein, mouse
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RNA, Small Interfering
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Wnt-5a Protein
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Wnt5a protein, mouse
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beta Catenin
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Casp3 protein, mouse
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Caspase 3