{"title":"抑制 PDHα1 基因表达对脂肪肝细胞代谢的影响","authors":"Xiaoguang Chen, Qiongxia Lv, Haonan Li, Zhe Wang, Yuxin Chang","doi":"10.1016/j.crbiot.2023.100174","DOIUrl":null,"url":null,"abstract":"<div><p>PDHα1 gene encodes catalytic subunit PDHE1α of pyruvate dehydrogenase (PDH) complex. Based on previous studies, it is hypothesized that inhibition of PDH activity prevents the entry of glycolytic pyruvate into TCA cycle, while promotes fatty acid oxidation and reduces liver triglyceride (TG) level, thereby alleviating nonalcoholic fatty liver disease (NAFLD). In this study, an in vitro model for NAFLD was established with medical fat emulsion; the most effective siRNA for PDHα1 gene was screened out by qPCR technology; the alterations in metabolism of glucose and lipid, and structure & function of mitochondria in the NAFLD cells were primarily evaluated after transfecting PDHα1 siRNA. As the results showed, after inhibiting the expression of PDHα1 gene, glucose level in culture medium was time-dependently increased, and LDH activity in the cells was moderately elevated after 24 h of transfection and then returned to the normal level after 48 h; intracellular TG level was decreased while LPS activity was increased in a time-dependent manner; no significant change in mitochondrial structure was observed with or without siRNA transfection, and ATP content was obviously reduced after 24 h of transfection, followed by restoration after 48 h. It can be concluded that inhibiting PDHα1 gene in fatty liver cells enhances lipid degradation, and represses the utilization of glucose to an extent, thus reducing TG level without impacting energy generation required for cell survival.</p></div>","PeriodicalId":52676,"journal":{"name":"Current Research in Biotechnology","volume":null,"pages":null},"PeriodicalIF":3.6000,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2590262823000564/pdfft?md5=1d838338247b96776cf94a22e4247851&pid=1-s2.0-S2590262823000564-main.pdf","citationCount":"0","resultStr":"{\"title\":\"Effect of inhibiting PDHα1 gene expression on the metabolism of fatty liver cells\",\"authors\":\"Xiaoguang Chen, Qiongxia Lv, Haonan Li, Zhe Wang, Yuxin Chang\",\"doi\":\"10.1016/j.crbiot.2023.100174\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>PDHα1 gene encodes catalytic subunit PDHE1α of pyruvate dehydrogenase (PDH) complex. Based on previous studies, it is hypothesized that inhibition of PDH activity prevents the entry of glycolytic pyruvate into TCA cycle, while promotes fatty acid oxidation and reduces liver triglyceride (TG) level, thereby alleviating nonalcoholic fatty liver disease (NAFLD). In this study, an in vitro model for NAFLD was established with medical fat emulsion; the most effective siRNA for PDHα1 gene was screened out by qPCR technology; the alterations in metabolism of glucose and lipid, and structure & function of mitochondria in the NAFLD cells were primarily evaluated after transfecting PDHα1 siRNA. As the results showed, after inhibiting the expression of PDHα1 gene, glucose level in culture medium was time-dependently increased, and LDH activity in the cells was moderately elevated after 24 h of transfection and then returned to the normal level after 48 h; intracellular TG level was decreased while LPS activity was increased in a time-dependent manner; no significant change in mitochondrial structure was observed with or without siRNA transfection, and ATP content was obviously reduced after 24 h of transfection, followed by restoration after 48 h. 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引用次数: 0
摘要
PDHα1 基因编码丙酮酸脱氢酶(PDH)复合物的催化亚基 PDHE1α。根据以往的研究,人们推测抑制 PDH 活性可阻止糖酵解丙酮酸进入 TCA 循环,同时促进脂肪酸氧化,降低肝脏甘油三酯(TG)水平,从而缓解非酒精性脂肪肝(NAFLD)。本研究利用医用脂肪乳剂建立了非酒精性脂肪肝的体外模型,通过 qPCR 技术筛选出了对 PDHα1 基因最有效的 siRNA,并主要评估了转染 PDHα1 siRNA 后非酒精性脂肪肝细胞中糖、脂代谢及线粒体结构和功能的改变。结果显示,抑制 PDHα1 基因表达后,培养液中葡萄糖水平呈时间依赖性升高,转染 24 h 后细胞中 LDH 活性中度升高,48 h 后恢复正常水平;细胞内 TG 水平下降,LPS 活性升高,且呈时间依赖性;转染与否线粒体结构无明显变化,转染 24 h 后 ATP 含量明显降低,48 h 后恢复正常。由此可以得出结论:抑制脂肪肝细胞中的 PDHα1 基因可促进脂质降解,并在一定程度上抑制葡萄糖的利用,从而在不影响细胞生存所需能量生成的情况下降低 TG 水平。
Effect of inhibiting PDHα1 gene expression on the metabolism of fatty liver cells
PDHα1 gene encodes catalytic subunit PDHE1α of pyruvate dehydrogenase (PDH) complex. Based on previous studies, it is hypothesized that inhibition of PDH activity prevents the entry of glycolytic pyruvate into TCA cycle, while promotes fatty acid oxidation and reduces liver triglyceride (TG) level, thereby alleviating nonalcoholic fatty liver disease (NAFLD). In this study, an in vitro model for NAFLD was established with medical fat emulsion; the most effective siRNA for PDHα1 gene was screened out by qPCR technology; the alterations in metabolism of glucose and lipid, and structure & function of mitochondria in the NAFLD cells were primarily evaluated after transfecting PDHα1 siRNA. As the results showed, after inhibiting the expression of PDHα1 gene, glucose level in culture medium was time-dependently increased, and LDH activity in the cells was moderately elevated after 24 h of transfection and then returned to the normal level after 48 h; intracellular TG level was decreased while LPS activity was increased in a time-dependent manner; no significant change in mitochondrial structure was observed with or without siRNA transfection, and ATP content was obviously reduced after 24 h of transfection, followed by restoration after 48 h. It can be concluded that inhibiting PDHα1 gene in fatty liver cells enhances lipid degradation, and represses the utilization of glucose to an extent, thus reducing TG level without impacting energy generation required for cell survival.
期刊介绍:
Current Research in Biotechnology (CRBIOT) is a new primary research, gold open access journal from Elsevier. CRBIOT publishes original papers, reviews, and short communications (including viewpoints and perspectives) resulting from research in biotechnology and biotech-associated disciplines.
Current Research in Biotechnology is a peer-reviewed gold open access (OA) journal and upon acceptance all articles are permanently and freely available. It is a companion to the highly regarded review journal Current Opinion in Biotechnology (2018 CiteScore 8.450) and is part of the Current Opinion and Research (CO+RE) suite of journals. All CO+RE journals leverage the Current Opinion legacy-of editorial excellence, high-impact, and global reach-to ensure they are a widely read resource that is integral to scientists' workflow.