Zhang Zhi-Mei, Liu Zhi-Hong, Nie Qian, Zhang Xue-Mei, Yang Li-Qun, Wang Chao, Yang Lin-Lin, Song Guang-Yao
Department of Internal Medicine, Hebei Medical University, Shijiazhuang, Hebei 050017, P.R. China.
Department of Endocrinology, Hebei General Hospital, Shijiazhuang, Hebei 050051, P.R. China.
Exp Ther Med. 2022 May;23(5):332. doi: 10.3892/etm.2022.11261. Epub 2022 Mar 16.
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 and (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 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 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.
二甲双胍（MET）是2型糖尿病患者的一线治疗选择，近年来受到了广泛关注。然而，评估其对胰岛素抵抗和长链非编码RNA（lncRNA）表达谱影响的研究数量不足。本研究将小鼠分为三组：对照组、高脂饮食（HFD）组和HFD + MET组。进行高通量测序分析以检测lncRNA和mRNA表达水平，并筛选出差异表达的lncRNA。随后，在胰岛素抵抗的[具体模型1]和[具体模型2]（用棕榈酸处理的小鼠肝脏AML12细胞）模型中对差异表达的lncRNA进行验证。通过逆转录定量PCR验证随机选择的lncRNA后，鉴定出一种新的lncRNA，NONMMUT031874.2，其在HFD组中上调，并通过MET治疗逆转。为了研究NONMMUT031874.2的下游机制，构建了lncRNA-微小RNA（miR/miRNA）-mRNA共表达网络，并使用了NONCODE、miRBase和TargetScan数据库，结果表明NONMMUT031874.2可能通过miR-7054-5p调节细胞因子信号转导抑制因子3。在本研究的[具体部分]中，在用棕榈酸（PA）和MET处理之前，用小干扰RNA转染AML12细胞以敲低NONMMUT031874.2的表达。结果表明，PA处理显著增加了NONMMUT031874.2的表达，而miR-7054-5p的表达显著降低。相比之下，敲低NONMMUT031874.2的表达或用MET处理后，上述观察结果被逆转。此外，还发现敲低NONMMUT031874.2和用MET处理在减轻AML12细胞胰岛素抵抗和降低葡萄糖浓度方面具有相似的效果。这些结果表明，MET治疗可通过下调NONMMUT031874.2的表达来改善胰岛素抵抗。
