Regulation of lipid droplet (LD) formation in hepatocytes via regulation of SREBP1c by non-coding RNAs

Theodor Bilharz Research Institute

Bibliographic Information

Authors: El Sobky S.A.; Aboud N.K.; El Assaly N.M.; Fawzy I.O.; El-Ekiaby N.; Abdelaziz A.I.

Journal: Frontiers in Medicine

Publisher: Frontiers Media S.A.

Publication Date: 20 September 2022

Volume / Issue: Volume 9

Article No.: 903856

ISSN: 2296858X

DOI: 10.3389/fmed.2022.903856

Scopus: View on Scopus

Document Type: Article

Access: All Open Access; Gold Open Access; Green Open Access

Authors and Affiliations

El Sobky S.A., School of Medicine, Newgiza University (NGU), Giza, Egypt; Aboud N.K., School of Medicine, Newgiza University (NGU), Giza, Egypt; El Assaly N.M., Clinical Chemistry Department, Theodor Bilharz Research Institute, Giza, Egypt; Fawzy I.O., School of Medicine, Newgiza University (NGU), Giza, Egypt; El-Ekiaby N., School of Medicine, Newgiza University (NGU), Giza, Egypt; Abdelaziz A.I., School of Medicine, Newgiza University (NGU), Giza, Egypt

Abstract

Introduction: Increased de novo lipogenesis (DNL) is one of the key factors contributing to fat accumulation and non-alcoholic fatty liver disease (NAFLD). Among the critical transcription factors (TFs) regulating DNL is mTOR and its downstream lipogenic TF, SREBP1c. In recent years, it has been established that non-coding RNAs (ncRNAs) play role in both biological processes and disease pathogenesis. Our group has previously characterized microRNAs that can target and regulate the expression of both mTOR and SREBP1c. Accordingly, this study aimed to broaden our understanding of the role of ncRNAs in regulating the mTOR/SREBP1c axis to elucidate the role of the non-coding transcriptome in DNL and lipid droplet (LD) formation. Hence, short ncRNA, miR-615-5p, and long non-coding RNA (lncRNA), H19, were chosen as they were previously proven to target mTOR by our group and in the published literature, respectively. Methodology: Huh-7 cells were treated with 800 μM oleic acid (OA) to promote LD formation. Transfection of miR-615-5p mimics or H19 over-expression vectors was performed, followed by the measurement of their downstream targets, mTOR and SREBP, on the mRNA level by quantitative real-time PCR (qRT-PCR), and on the protein level by Western blot. To determine the functional impact of miR-615-5p and H19 on LD formation and triglyceride (TG) accumulation, post-transfection LDs were stained, imaged, and characterized, and TGs were extracted and quantified. Results: miR-615-5p was able to reduce mTOR and SREBP1c significantly on both the mRNA and protein levels compared to control cells, while H19 caused a reduction of both targets on the protein level only. Both miR-615-5p and H19 were able to significantly reduce the LD count and total area, as well as TG levels compared to control cells. Conclusion: To conclude, this study shows, for the first time, the impact of miR-615-5p and H19 on the mTOR/SREBP1c axis, and thus, their functional impact on LDs and TG accumulation. These findings might pave the way for using ncRNAs as potential therapeutic targets in the management of fatty liver. Copyright © 2022 El Sobky, Aboud, El Assaly, Fawzy, El-Ekiaby and Abdelaziz.

Keywords

de novo lipogenesis (DNL); mTOR; NAFLD; ncRNAs; SREBP1c; complementary DNA; fat droplet; long untranslated RNA; mammalian target of rapamycin; messenger RNA; microRNA; microRNA 615 5p; microRNA h19; sterol regulatory element binding protein; sterol regulatory element binding protein 1c; transcriptome; triacylglycerol; unclassified drug; untranslated RNA; Article; cell viability; controlled study; cytotoxicity; expression vector; fatty liver; gene expression; genetic transcription; genetic transfection; Huh-7 cell line; human; human cell; interphase; lipogenesis; liver cell; nonalcoholic fatty liver; protein expression; protein expression level; protein function; quantitative analysis; real time polymerase chain reaction; reverse transcription polymerase chain reaction; triacylglycerol blood level; triacylglycerol level; Western blotting

Citation Information

Scopus Citations: 9

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