PROFILE OF LONG NON-CODING RNA IN RESPONSE TO SULFORAPHANE ANTI-OBESITY TREATMENT IN SKELETAL MUSCLES OF DIET INDUCED OBESITY MICE

Abstract

Background: World Health Organization data reveal a rapid growth in obesity worldwide, making it a major public health concern. Obesity has been studied extensively in recent decades; however, it is still challenging due to complex interactions of genetic and environmental factors. Long noncoding RNAs (lncRNAs) are emerging as novel regulators linked to obesity and obesity-associated complications. Herbal chemicals like Sulforaphane (SFN) isolated from green vegetables can alleviate obesity. Recent findings suggest that SFN can prevent diet- induced obesity. Aim: To examine the changes in lncRNAs transcripts/ target mRNAs and their effects on gene ontology (GO) and signaling pathways in skeletal muscles (SMs) of diet- induced obese (DIO) mice following SFN treatment. Methods: The DIO mice models were constructed by constant feeding with a high-fat diet. Then they were divided into two groups. The first group was treated with SFN for four weeks, while the second group was treated with a vehicle for the same time period. The body weight and food intake were measured during the four weeks. In addition, the glucose tolerance test (GTT) was performed and, serum glucose, leptin, and insulin were assessed. Total RNA was extracted from SM tissues, and lncRNA sequencing (RNA-seq) was performed to identify differentially expressed transcripts of lncRNAs and their target mRNA in response to SFN treatment. In addition, bioinformatic analysis, and GO enrichment analysis and pathway analysis were performed to identify differentially expressed lncRNA/mRNA. Results: SFN significantly reduced the body weight and cumulative food intake of DIO mice. Moreover, treatment with SFN markedly lowered the biochemical parameters, including blood glucose and leptin, and improved the glucose uptake by SMs. In response to SFN treatment, RNA-seq analysis of the SMs of DIO mice identified 30 differentially expressed lncRNAs (9 upregulated and 21 downregulated), and 610 differentially expressed mRNA target genes (101 upregulated and 509 downregulated). Furthermore, the differentially expressed lncRNA transcript and mRNA target genes discovered in this study were primarily associated with insulin resistance, metabolic pathways, lipid metabolism, Tumor Necrosis Factor (TNF), and Toll-like receptor (TLR) signaling pathways. Conclusion: SFN successfully reduced obesity by lowering body weight and total calorie intake. Profiling lncRNA and mRNA in the SMs of DIO mice revealed various differentially expressed lncRNAs and mRNAs. The enrichment analysis showed that the identified differentially expressed genes could be implicated in obesity by several pathways. Gene enrichment analysis suggested that signaling pathways and GO processes connected to SMs metabolism may be implicated, requiring further experimental validation.