MiR-495 regulates macrophage M1/M2 polarization and insulin resistance in high-fat diet-fed mice via targeting FTO
MicroRNA 495 (miR-495) has been discovered to be involved in the metabolism and immune response in human body. The purpose of this study was to investigate the effect of miR-495 on macrophage M1/M2 polarization and insulin resistance in type 2 diabetes (T2D). A T2D mouse model was established by feeding C57BL/6 mice with a high-fat diet (HFD). The expressions of M1/M2 polarization markers and miR-495 in peritoneal macrophages were determined by qRT-PCR or Western blot. Mouse insulin tolerance test (ITT) and glucose tolerance test (GTT) were performed, and the targeted binding effect between miR-495, fat mass, and obesity-associated gene (FTO) was verified by double luciferase gene reporter assay. The body weight, blood glucose content, and miR-495 expression in macrophages of the HFD group were remarkably higher than those of the normal diet (ND) group. Besides, miR-495 induced the transformation of macrophages into M1-type pro-inflammatory macrophages and enhanced the insulin resistance of T2D mice. More importantly, FTO was proved to be a direct target gene of miR-495 and silencing FTO could induce the transformation of macrophages into M1-type pro-inflammatory macrophages. These results demonstrated that miR-495 could promote the transformation of macrophages into M1-type pro-inflammatory macrophages by inhibiting the expression of its target gene FTO, and aggravate the insulin resistance and adipose tissue inflammation in T2D mice, which provided a certain theoretical basis for the targeted treatment of T2D.
KeywordsHigh-fat diet Type 2 diabetes miR-495 FTO Macrophage M1/M2 polarization
FH: study concepts, study design, literature research, experimental studies, manuscript preparation and editing; JKT: literature research experimental studies, manuscript preparation, and editing; BLD: experiments work, data acquisition, statistical analysis, and manuscript writing; JZ: data acquisition, statistical analysis, and clinical study; CZL: study concepts, definition of intellectual content, study design, data analysis, and manuscript review. All authors read and approved the final manuscript.
Compliance with ethical standards
Conflicts of interest
The authors declare that they have no conflict of interest.
Research involving Animals
Ethical approval was obtained from the Ethics Committee of Tianjin First Central Hospital. All procedures performed in studies involving animals were conducted in accordance with the animal care and guidelines of Tianjin First Central Hospital.
- 2.Benedict C, Axelsson T, Soderberg S, Larsson A, Ingelsson E, Lind L, Schioth HB (2014) Fat mass and obesity-associated gene (FTO) is linked to higher plasma levels of the hunger hormone ghrelin and lower serum levels of the satiety hormone leptin in older adults. Diabetes 63:3955–3959. https://doi.org/10.2337/db14-0470 CrossRefPubMedGoogle Scholar
- 3.Claussnitzer M, Dankel SN, Kim KH, Quon G, Meuleman W, Haugen C, Glunk V, Sousa IS, Beaudry JL, Puviindran V, Abdennur NA, Liu J, Svensson PA, Hsu YH, Drucker DJ, Mellgren G, Hui CC, Hauner H, Kellis M (2015) FTO obesity variant circuitry and adipocyte browning in humans. N Engl J Med 373:895–907. https://doi.org/10.1056/NEJMoa1502214 CrossRefPubMedPubMedCentralGoogle Scholar
- 5.Cuccarese MF, Dubach JM, Pfirschke C, Engblom C, Garris C, Miller MA, Pittet MJ, Weissleder R (2017) Heterogeneity of macrophage infiltration and therapeutic response in lung carcinoma revealed by 3D organ imaging. Nat Commun 8:14293. https://doi.org/10.1038/ncomms14293 CrossRefPubMedPubMedCentralGoogle Scholar
- 9.Du J, Paz K, Flynn R, Vulic A, Robinson TM, Lineburg KE, Alexander KA, Meng J, Roy S, Panoskaltsis-Mortari A, Loschi M, Hill GR, Serody JS, Maillard I, Miklos D, Koreth J, Cutler CS, Antin JH, Ritz J, MacDonald KP, Schacker TW, Luznik L, Blazar BR (2017) Pirfenidone ameliorates murine chronic GVHD through inhibition of macrophage infiltration and TGF-beta production. Blood 129:2570–2580. https://doi.org/10.1182/blood-2017-01-758854 CrossRefPubMedPubMedCentralGoogle Scholar
- 10.Dyson J, Jaques B, Chattopadyhay D, Lochan R, Graham J, Das D, Aslam T, Patanwala I, Gaggar S, Cole M, Sumpter K, Stewart S, Rose J, Hudson M, Manas D, Reeves HL (2014) Hepatocellular cancer: the impact of obesity, type 2 diabetes and a multidisciplinary team. J Hepatol 60:110–117. https://doi.org/10.1016/j.jhep.2013.08.011 CrossRefPubMedGoogle Scholar
- 12.Formosa A, Markert EK, Lena AM, Italiano D, Finazzi-Agro E, Levine AJ, Bernardini S, Garabadgiu AV, Melino G, Candi E (2014) MicroRNAs, miR-154, miR-299-5p, miR-376a, miR-376c, miR-377, miR-381, miR-487b, miR-485-3p, miR-495 and miR-654-3p, mapped to the 14q32.31 locus, regulate proliferation, apoptosis, migration and invasion in metastatic prostate cancer cells. Oncogene 33:5173–5182. https://doi.org/10.1038/onc.2013.451 CrossRefPubMedGoogle Scholar
- 13.Han Y-B, Tian M, Jin J, Zou G-L, Sui Y-B, Peng P, Liu L (2018) Berberine improves metabolic syndrome insulin resistance by inducing macrophage M2 polarization. Int J Clin Exp Med 11:11191–11197Google Scholar
- 15.Kawano Y, Nakae J, Watanabe N, Kikuchi T, Tateya S, Tamori Y, Kaneko M, Abe T, Onodera M, Itoh H (2016) Colonic pro-inflammatory macrophages cause insulin resistance in an intestinal Ccl2/Ccr2-dependent manner. Cell Metab 24:295–310. https://doi.org/10.1016/j.cmet.2016.07.009 CrossRefPubMedPubMedCentralGoogle Scholar
- 16.Khan IM, Perrard XY, Brunner G, Lui H, Sparks LM, Smith SR, Wang X, Shi ZZ, Lewis DE, Wu H, Ballantyne CM (2015) Intermuscular and perimuscular fat expansion in obesity correlates with skeletal muscle T cell and macrophage infiltration and insulin resistance. Int J Obes (Lond) 39:1607–1618. https://doi.org/10.1038/ijo.2015.104 CrossRefGoogle Scholar
- 24.Maan NS, Maan S, Belaganahalli M, Pullinger G, Montes AJA, Gasparini MR, Guimera M, Nomikou K, Mertens PP (2015) A quantitative real-time reverse transcription PCR (qRT-PCR) assay to detect genome segment 9 of all 26 bluetongue virus serotypes. Journal of virological methods 213:118–126CrossRefGoogle Scholar
- 25.Marso SP, Daniels GH, Brown-Frandsen K, Kristensen P, Mann JF, Nauck MA, Nissen SE, Pocock S, Poulter NR, Ravn LS, Steinberg WM, Stockner M, Zinman B, Bergenstal RM, Buse JB, Committee LS, Investigators LT (2016) Liraglutide and cardiovascular outcomes in type 2 diabetes. N Engl J Med 375:311–322. https://doi.org/10.1056/NEJMoa1603827 CrossRefPubMedPubMedCentralGoogle Scholar
- 26.Mingrone G, Panunzi S, De Gaetano A, Guidone C, Iaconelli A, Nanni G, Castagneto M, Bornstein S, Rubino F (2015) Bariatric-metabolic surgery versus conventional medical treatment in obese patients with type 2 diabetes: 5 year follow-up of an open-label, single-centre, randomised controlled trial. Lancet 386:964–973. https://doi.org/10.1016/S0140-6736(15)00075-6 CrossRefPubMedGoogle Scholar
- 27.Nilsson E, Jansson PA, Perfilyev A, Volkov P, Pedersen M, Svensson MK, Poulsen P, Ribel-Madsen R, Pedersen NL, Almgren P (2014) Altered DNA methylation and differential expression of genes influencing metabolism and inflammation in adipose tissue from subjects with type 2 diabetes. Diabetes 63:2962–2976CrossRefGoogle Scholar
- 30.Olivo G, Wiemerslage L, Nilsson EK, Solstrand Dahlberg L, Larsen AL, Olaya Bucaro M, Gustafsson VP, Titova OE, Bandstein M, Larsson EM, Benedict C, Brooks SJ, Schioth HB (2016) Resting-state brain and the FTO obesity risk allele: default mode, sensorimotor, and salience network connectivity underlying different somatosensory integration and reward processing between genotypes. Front Hum Neurosci 10:52. https://doi.org/10.3389/fnhum.2016.00052 CrossRefPubMedPubMedCentralGoogle Scholar
- 32.Pan Y, Hui X, Hoo RLC, Ye D, Chan CYC, Feng T, Wang Y, Lam KSL, Xu A (2019) Adipocyte-secreted exosomal microRNA-34a inhibits M2 macrophage polarization to promote obesity-induced adipose inflammation. J Clin Invest 129:834–849. https://doi.org/10.1172/JCI123069 CrossRefPubMedPubMedCentralGoogle Scholar
- 36.Ronkainen J, Huusko TJ, Soininen R, Mondini E, Cinti F, Makela KA, Kovalainen M, Herzig KH, Jarvelin MR, Sebert S, Savolainen MJ, Salonurmi T (2015) Fat mass- and obesity-associated gene Fto affects the dietary response in mouse white adipose tissue. Sci Rep 5:9233. https://doi.org/10.1038/srep09233 CrossRefPubMedPubMedCentralGoogle Scholar
- 40.Suzuki T, Gao J, Ishigaki Y, Kondo K, Sawada S, Izumi T, Uno K, Kaneko K, Tsukita S, Takahashi K, Asao A, Ishii N, Imai J, Yamada T, Oyadomari S, Katagiri H (2017) ER stress protein CHOP mediates insulin resistance by modulating adipose tissue macrophage polarity. Cell Rep 18:2045–2057. https://doi.org/10.1016/j.celrep.2017.01.076 CrossRefPubMedGoogle Scholar
- 43.van Diepen JA, Robben JH, Hooiveld GJ, Carmone C, Alsady M, Boutens L, Bekkenkamp-Grovenstein M, Hijmans A, Engelke UFH, Wevers RA, Netea MG, Tack CJ, Stienstra R, Deen PMT (2017) SUCNR1-mediated chemotaxis of macrophages aggravates obesity-induced inflammation and diabetes. Diabetologia 60:1304–1313. https://doi.org/10.1007/s00125-017-4261-z CrossRefPubMedPubMedCentralGoogle Scholar
- 45.Xu L, Nagata N, Nagashimada M, Zhuge F, Ni Y, Chen G, Mayoux E, Kaneko S, Ota T (2017) SGLT2 inhibition by empagliflozin promotes fat utilization and browning and attenuates inflammation and insulin resistance by polarizing M2 macrophages in diet-induced obese mice. EBioMedicine 20:137–149CrossRefGoogle Scholar
- 46.Zhuang G, Meng C, Guo X, Cheruku PS, Shi L, Xu H, Li H, Wang G, Evans AR, Safe S, Wu C, Zhou B (2012) A novel regulator of macrophage activation: miR-223 in obesity-associated adipose tissue inflammation. Circulation 125:2892–2903. https://doi.org/10.1161/CIRCULATIONAHA.111.087817 CrossRefPubMedGoogle Scholar
- 48.Zinman B, Wanner C, Lachin JM, Fitchett D, Bluhmki E, Hantel S, Mattheus M, Devins T, Johansen OE, Woerle HJ, Broedl UC, Inzucchi SE, Investigators E-RO (2015) Empagliflozin, cardiovascular outcomes, and mortality in type 2 diabetes. N Engl J Med 373:2117–2128. https://doi.org/10.1056/NEJMoa1504720 CrossRefPubMedPubMedCentralGoogle Scholar