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

Zhang,Zhi-Mei; Liu,Zhi-Hong; Nie,Qian; Zhang,Xue-Mei; Yang,Li-Qun; Wang,Chao; Yang,Lin-Lin; Song,Guang-Yao

doi:10.3892/etm.2022.11261

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

Authors:
- Zhi-Mei Zhang
- Zhi-Hong Liu
- Qian Nie
- Xue-Mei Zhang
- Li-Qun Yang
- Chao Wang
- Lin-Lin Yang
- Guang-Yao Song
View Affiliations

Affiliations: Department of Internal Medicine, Hebei Medical University, Shijiazhuang, Hebei 050017, P.R. China, Hebei Key Laboratory of Metabolic Diseases, Hebei General Hospital, Shijiazhuang, Hebei 050051, P.R. China
Published online on: March 16, 2022 https://doi.org/10.3892/etm.2022.11261
Article Number: 332
Copyright: © Zhang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

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.

View Figures

View References

1	Ning G: Decade in review-type 2 diabetes mellitus: At the centre of things. Nat Rev Endocrinol. 11:636–638. 2015.PubMed/NCBI View Article : Google Scholar
2	Ogurtsova K, da Rocha Fernandes JD, Huang Y, Linnenkamp U, Guariguata L, Cho NH, Cavan D, Shaw JE and Makaroff LE: IDF diabetes atlas: Global estimates for the prevalence of diabetes for 2015 and 2040. Diabetes Res Clin Pract. 128:40–50. 2017.PubMed/NCBI View Article : Google Scholar
3	Akash MSH, Rehman K and Liaqat A: Tumor necrosis factor-alpha: Role in development of insulin resistance and pathogenesis of type 2 diabetes mellitus. J Cell Biochem. 119:105–110. 2018.PubMed/NCBI View Article : Google Scholar
4	Kurokawa R, Rosenfeld MG and Glass CK: Transcriptional regulation through noncoding RNAs and epigenetic modifications. RNA Biol. 6:233–236. 2009.PubMed/NCBI View Article : Google Scholar
5	Ravasi T, Suzuki H, Pang KC, Katayama S, Furuno M, Okunishi R, Fukuda S, Ru K, Frith MC, Gongora MM, et al: Experimental validation of the regulated expression of large numbers of non-coding RNAs from the mouse genome. Genome Res. 16:11–19. 2006.PubMed/NCBI View Article : Google Scholar
6	Sathishkumar C, Prabu P, Mohan V and Balasubramanyam M: Linking a role of lncRNAs (long non-coding RNAs) with insulin resistance, accelerated senescence, and inflammation in patients with type 2 diabetes. Hum Genomics. 12(41)2018.PubMed/NCBI View Article : Google Scholar
7	Hanson A, Wilhelmsen D and DiStefano JK: The role of long non-coding RNAs (lncRNAs) in the development and progression of fibrosis associated with nonalcoholic fatty liver disease (NAFLD). Noncoding RNA. 4(18)2018.PubMed/NCBI View Article : Google Scholar
8	Pielok A and Marycz K: Non-coding RNAs as potential novel biomarkers for early diagnosis of hepatic insulin resistance. Int J Mol Sci. 21(4182)2020.PubMed/NCBI View Article : Google Scholar
9	Davies MJ, D'Alessio DA, Fradkin J, Kernan WN, Mathieu C, Mingrone G, Rossing P, Tsapas A, Wexler DJ and Buse JB: Management of hyperglycaemia in type 2 diabetes, 2018. A consensus report by the American diabetes association (ADA) and the European association for the study of diabetes (EASD). Diabetologia. 61:2461–2498. 2018.PubMed/NCBI View Article : Google Scholar
10	Hundal RS, Krssak M, Dufour S, Laurent D, Lebon V, Chandramouli V, Inzucchi SE, Schumann WC, Petersen KF, Landau BR and Shulman GI: Mechanism by which metformin reduces glucose production in type 2 diabetes. Diabetes. 49:2063–2069. 2000.PubMed/NCBI View Article : Google Scholar
11	Ren GF, Xiao LL, Ma XJ, Yan YS and Jiao PF: Metformin decreases insulin resistance in type 1 diabetes through regulating p53 and RAP2A in vitro and in vivo. Drug Des Devel Ther. 14:2381–2392. 2020.PubMed/NCBI View Article : Google Scholar
12	Grycel S, Markowski AR, Hady HR, Zabielski P, Kojtallmierska M, Górski J and Blachnio-Zabielska AU: Metformin treatment affects adipocytokine secretion and lipid composition in adipose tissues of diet-induced insulin-resistant rats. Nutrition. 63-64:126–133. 2019.PubMed/NCBI View Article : Google Scholar
13	Wang Y, Tang H, Ji X, Zhang Y, Xu W, Yang X, Deng R, Liu Y, Li F, Wang X and Zhou L: Expression profile analysis of long non-coding RNAs involved in the metformin-inhibited gluconeogenesis of primary mouse hepatocytes. Int J Mol Med. 41:302–310. 2018.PubMed/NCBI View Article : Google Scholar
14	Ferdowsian H: Human and animal research guidelines: Aligning ethical constructs with new scientific developments. Bioethics. 25:472–478. 2011.PubMed/NCBI View Article : Google Scholar
15	Zhao H, Zhang Y, Shu L, Song G and Ma H: Resveratrol reduces liver endoplasmic reticulum stress and improves insulin sensitivity in vivo and in vitro. Drug Des Devel Ther. 13:1473–1485. 2019.PubMed/NCBI View Article : Google Scholar
16	Katz A, Nambi SS, Mather K, Baron AD, Follmann DA, Sullivan G and Quon MJ: Quantitative insulin sensitivity check index: A simple, accurate method for assessing insulin sensitivity in humans. J Clin Endocrinol Metab. 85:2402–2410. 2000.PubMed/NCBI View Article : Google Scholar
17	Fang F, Li H, Qin T, Li M and Ma S: Thymol improves high-fat diet-induced cognitive deficits in mice via ameliorating brain insulin resistance and upregulating NRF2/HO-1 pathway. Metab Brain Dis. 32:385–393. 2017.PubMed/NCBI View Article : Google Scholar
18	Zhang R, Chu K, Zhao N, Wu J, Ma L, Zhu C, Chen X, Wei G and Liao M: Corilagin alleviates nonalcoholic fatty liver disease in high-fat diet-induced C57BL/6 mice by ameliorating oxidative stress and restoring autophagic flux. Front Pharmacol. 10(1693)2020.PubMed/NCBI View Article : Google Scholar
19	Villalva-Pérez JM, Ramírez-Vargas MA, Serafín-Fabían JI, Ramírez M, Moreno-Godínez ME, Espinoza-Rojo M and Flores-Alfaro E: Characterization of Huh7 cells after the induction of insulin resistance and post-treatment with metformin. Cytotechnology. 72:499–511. 2020.PubMed/NCBI View Article : Google Scholar
20	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-delta C(T)) method. Methods. 25:402–408. 2001.PubMed/NCBI View Article : Google Scholar
21	Robinson MD, McCarthy DJ and Smyth GK: edgeR: A bioconductor package for differential expression analysis of digital gene expression data. Bioinformatics. 26:139–140. 2010.PubMed/NCBI View Article : Google Scholar
22	Moriya Y, Itoh M, Okuda S, Yoshizawa A and Kanehisa M: KAAS: An automatic genome annotation and pathway reconstruction server. Nucleic Acids Res. 35 (Web Server Issue):W182–W185. 2007.PubMed/NCBI View Article : Google Scholar
23	Rice P, Longden I and Bleasby A: EMBOSS: The European molecular biology open software suite. Trends Genet. 16:276–277. 2000.PubMed/NCBI View Article : Google Scholar
24	Li YC, Kang L, Huang J, Zhang J, Liu C and Shen W: Effects of miR-152-mediated targeting of SOCS3 on hepatic insulin resistance in gestational diabetes mellitus mice. Am J Med Sci. 361:365–374. 2021.PubMed/NCBI View Article : Google Scholar
25	Guariguata L, Whiting DR, Hambleton I, Beagley J, Linnenkamp U and Shaw JE: Global estimates of diabetes prevalence for 2013 and projections for 2035. Diabetes Res Clin Pract. 103:137–149. 2014.PubMed/NCBI View Article : Google Scholar
26	Kim J, Bilder D and Neufeld TP: Mechanical stress regulates insulin sensitivity through integrin-dependent control of insulin receptor localization. Genes Dev. 32:156–164. 2018.PubMed/NCBI View Article : Google Scholar
27	Yan C, Chen J and Chen N: Long noncoding RNA MALAT1 promotes hepatic steatosis and insulin resistance by increasing nuclear SREBP-1c protein stability. Sci Rep. 6(22640)2016.PubMed/NCBI View Article : Google Scholar
28	Shu L, Hou G, Zhao H, Huang W, Song G and Ma H: Resveratrol improves high-fat diet-induced insulin resistance in mice by downregulating the lncRNA NONMMUT008655.2. Am J Transl Res. 12:1–18. 2020.PubMed/NCBI
29	Yin DD, Zhang EB, You LH, Wang N, Wang LT, Jin FY, Zhu YN, Cao LH, Yuan QX, De W and Tang W: Downregulation of lncRNA TUG1 affects apoptosis and insulin secretion in mouse pancreatic β cells. Cell Physiol Biochem. 35:1892–1904. 2015.PubMed/NCBI View Article : Google Scholar
30	Yang Q, Zhu Z, Wang L, Xia H, Mao J, Wu J, Kato K, Li H, Zhang J, Yamanaka K and An Y: The protective effect of silk fibroin on high glucose induced insulin resistance in HepG2 cells. Environ Toxicol Pharmacol. 69:66–71. 2019.PubMed/NCBI View Article : Google Scholar
31	Choi BR, Kim HJ, Lee YJ and Ku SK: Anti-diabetic obesity effects of Wasabia japonica matsum leaf extract on 45% kcal high-fat diet-fed mice. Nutrients. 12(2837)2020.PubMed/NCBI View Article : Google Scholar
32	Cao L, Wang Z and Wan W: Suppressor of cytokine signaling 3: Emerging role linking central insulin resistance and Alzheimer's disease. Front Neurosci. 12(417)2018.PubMed/NCBI View Article : Google Scholar
33	Galic S, Sachithanandan N, Kay TW and Steinberg GR: Suppressor of cytokine signalling (SOCS) proteins as guardians of inflammatory responses critical for regulating insulin sensitivity. Biochem J. 461:177–188. 2014.PubMed/NCBI View Article : Google Scholar
34	Song C, Liu D, Yang S, Cheng L, Xing E and Chen Z: Sericin enhances the insulin-PI3K/AKT signaling pathway in the liver of a type 2 diabetes rat model. Exp Ther Med. 16:3345–3352. 2018.PubMed/NCBI View Article : Google Scholar
35	Yang P, Liang Y, Luo Y, Li Z, Wen Y, Shen J, Li R, Zheng H, Gu HF and Xia N: Liraglutide ameliorates nonalcoholic fatty liver disease in diabetic mice via the IRS2/PI3K/Akt signaling pathway. Diabetes Metab Syndr Obes. 12:1013–1021. 2019.PubMed/NCBI View Article : Google Scholar
36	Xu L, Li Y, Yin L, Qi Y, Sun H, Sun P, Xu M, Tang Z and Peng J: miR-125a-5p ameliorates hepatic glycolipid metabolism disorder in type 2 diabetes mellitus through targeting of STAT3. Theranostics. 8:5593–5609. 2018.PubMed/NCBI View Article : Google Scholar
37	Yang Z, Huang W, Zhang J, Xie M and Wang X: Baicalein improves glucose metabolism in insulin resistant HepG2 cells. Eur J Pharmacol. 854:187–193. 2019.PubMed/NCBI View Article : Google Scholar
38	Cui X, Qian DW, Jiang S, Shang EX, Zhu ZH and Duan JA: Scutellariae radix and coptidis rhizoma improve glucose and lipid metabolism in T2DM rats via regulation of the metabolic profiling and MAPK/PI3K/Akt signaling pathway. Int J Mol Sci. 19(3634)2018.PubMed/NCBI View Article : Google Scholar
39	Ma H, Yuan J, Ma J, Ding J, Lin W, Wang X, Zhang M, Sun Y, Wu R, Liu C, et al: BMP7 improves insulin signal transduction in the liver via inhibition of mitogen-activated protein kinases. J Endocrinol. 243:97–110. 2019.PubMed/NCBI View Article : Google Scholar
40	Han M, You L, Wu Y, Gu N, Wang Y, Feng X, Xiang L, Chen Y, Zeng Y and Zhong T: RNA-sequencing analysis reveals the potential contribution of lncRNAs in palmitic acid-induced insulin resistance of skeletal muscle cells. Biosci Rep. 40(BSR20192523)2020.PubMed/NCBI View Article : Google Scholar
41	Chen DL, Shen DY, Han CK and Tian Y: LncRNA MEG3 aggravates palmitate-induced insulin resistance by regulating miR-185-5p/Egr2 axis in hepatic cells. Eur Rev Med Pharmacol Sci. 23:5456–5467. 2019.PubMed/NCBI View Article : Google Scholar
42	Fan CN, Ma L and Liu N: Systematic analysis of lncRNA-miRNA-mRNA competing endogenous RNA network identifies four-lncRNA signature as a prognostic biomarker for breast cancer. J Transl Med. 16(264)2018.PubMed/NCBI View Article : Google Scholar
43	Zhu X, Li H, Wu Y, Zhou J, Yang G and Wang W: lncRNA MEG3 promotes hepatic insulin resistance by serving as a competing endogenous RNA of miR-214 to regulate ATF4 expression. Int J Mol Med. 43:345–357. 2019.PubMed/NCBI View Article : Google Scholar