Metformin improves high‑fat diet‑induced insulin resistance in mice by downregulating the expression of long noncoding RNA NONMMUT031874.2

Abstract

Metformin (MET) is the first-line therapeutic option for patients with type 2 diabetes that has garnered substantial attention over recent years. However, an insufficient number of studies have been performed to assess its effects on insulin resistance and the expression profile of long noncoding RNAs (lncRNAs). The present study divided mice into three groups: Control group, high-fat diet (HFD) group and HFD + MET group. A high-throughput sequencing analysis was conducted to detect lncRNA and mRNA expression levels, and differentially expressed lncRNAs were selected. Subsequently, the differentially expressed lncRNAs were validated both in vivo and in vitro (mouse liver AML12 cells treated with Palmitic acid) models of insulin resistance. After validating randomly selected lncRNAs via reverse transcription-quantitative PCR a novel lncRNA, NONMMUT031874.2, was identified, which was upregulated in the HFD group and reversed with MET treatment. To investigate the downstream mechanism of NONMMUT031874.2, lncRNA-microRNA (miR/miRNA)-mRNA co-expression network was constructed and NONCODE, miRBase and TargetScan databases were used, which indicated that NONMMUT031874.2 may regulate suppressor of cytokine signaling 3 by miR-7054-5p. For the in vitro part of the present study, AML12 cells were transfected with small interfering RNA to knock down NONMMUT031874.2 expression before being treated with palmitic acid (PA) and MET. The results showed that the expression of NONMMUT031874.2 was significantly increased whereas miR-7054-5p expression was significantly decreased by PA treatment. By contrast, after knocking down NONMMUT031874.2 expression or treatment with MET, the aforementioned in vitro observations were reversed. In addition, it was also found that NONMMUT031874.2 knockdown and treatment with MET exerted similar effects in alleviating insulin resistance and whilst decreasing glucose concentration in AML12 cells. These results suggest that MET treatment can ameliorate insulin resistance by downregulating NONMMUT031874.2 expression.

IRW (Isoleucine–Arginine–Tryptophan) Improves Glucose Tolerance in High Fat Diet Fed C57BL/6 Mice via Activation of Insulin Signaling and AMPK Pathways in Skeletal Muscle

“…For example, AdipoRon, a small-molecule agonist of adiponectin receptor is used at 50 mg/kg BW [ 29 ], curcumin at 100 mg/kg BW [ 30 ], soy β-Conglycin at 10% of diet ( w / w ) [ 31 ] and the di-peptide (Trp-His) at dosages ranging between 10–100 mg/kg BW [ 32 , 33 ]. Moreover, metformin is used at 200 mg/kg BW in rodent models of diabetes [ 34 , 35 ].…”

Metformin Treatment Modulates Long Non-Coding RNA Isoforms Expression in Human Cells

“…Mice lncRNAs have been previously associated with metformin treatment accessed by RT-qPCR and microarray analyses [ 74 , 75 , 76 ]. Nevertheless, these types of analyses do not allow isoform-specific evaluation due to technical limitations and also because of the less extensive ncRNA annotation of mouse if compared to human genome.…”

Role of maraviroc and/or rapamycin in the liver of IL10 KO mice with frailty syndrome

“…The concentration of total protein in each sample was determined by the Bradford method. 5´adenosine monophosphate-activated protein kinase (AMPK) [58], phosphorylated AMPK (pAMPK) [59], protein kinase-B (Akt) [60], phosphorylated Akt (pAkt) [61], nuclear factor-kB (NF-kB) [62], phosphorylated NF-kB (pNF-kB) [63], mammalian target of (mTOR) [64], phosphorylated mTOR (pmTOR) [65], signal transducer and activator of transcription 3 (STAT3) [66] and phosphorylated STAT3 [66] were evaluated by Western blotting (WB). GAPDH was used as an internal control [67] S2 Table . Proteins were analyzed by colorimetry using a secondary antibody bound to peroxidase (anti-rabbit or mouse IgG, Cell Signaling, Danvers, MA) and incubated with the corresponding substrate [28].…”