目的:探讨小鼠脂肪前体细胞成棕色脂肪能力与增龄的相关性及其分子机制。方法:体外培养不同年龄段小鼠的脂肪前体细胞,采用油红O染色鉴定前体细胞成棕脂能力;采用实时定量PCR检测年轻组和年老组小鼠脂肪前体细胞中微小RNA(micro RNA,miR)-146b-3p表达情况;通过转染miR-146b-3p模拟物或抑制物,使其过表达或抑制其表达,实时定量PCR法检测相关的脂联素、脂滴包被蛋白、脂肪酸结合蛋白4、解偶联蛋白-1基因的表达情况。结果:年老组小鼠的脂肪前体细胞成棕脂分化能力较年轻组小鼠明显减弱,同时年老组小鼠的脂肪前体细胞miR-146b-3p表达也下降。过表达miR-146b-3p可改善年老组小鼠脂肪前体细胞的成棕脂能力,而年轻组小鼠脂肪前体细胞的成脂能力随miR-146b-3p表达下降而降低。结论:脂肪前体细胞向棕色脂肪细胞分化的能力随着年龄的增加而降低,miR-146b-3p可能是其中的一个作用靶点。
Objective: To investigate correlation between brown adipogeneic capacity of preadipocytes and ageing and the related mechanism in mice. Methods: Preadipocytes derived from various age mice were cultured in vitro, and oil red O staining was used to identify the brown adipogenic capacity. The expression of miR-146b-3p in young and old preadipocytes was detected by real-time quantitative PCR. Then miR-146b-3p inhibitors were transfected into young preadipocytes, while miR-146b-3p mimics were transfected into the old, and the relative mRNA levels of PLIN, FABP4, UCP-1, APN were measured by real-time quantitative PCR. Results: Preadipocytes of aged mice showed a weaker capacity of brown adipogenic capacity than that of the young mice, and the expression of miR-146b-3p in the preadipocytes of the aged group was also decreased. Overexpression of miR-146b-3p can improve the brown adipogenic capacity of preadipocytes in aged mice. However, the brown adipogenic capacity of the young declined with the decrease of miR-146b-3p expression. Conclusions: The brown adipogenic capacity of preadipocytes decreases with age, and miR-146b-3p might be one of the targets.
[1] Peirce V, Carobbio S, Vidal-Puig A.The different shades of fat[J]. Nature,2014,510(7503):76-83.
[2] Cannon B, Nedergaard J.Brown adipose tissue: function and physiological significance[J]. Physiol Rev, 2004,84(1):277-359.
[3] Villarroya F, Cereijo R, Villarroya J, et al.Brown adipose tissue as a secretory organ[J]. Nat Rev Endocrinol,2017, 13(1):26-35.
[4] Stanford KI, Middelbeek RJ, Townsend KL, et al.Brown adipose tissue regulates glucose homeostasis and insulin sensitivity[J]. J Clin Invest,2013,123(1):215-223.
[5] Zingaretti M1, Crosta F, Vitali A, et al. The presence of UCP1 demonstrates that metabolically active adipose tissue in the neck of adult humans truly represents brown adipose tissue[J]. FASEB J, 2009, 23(9):3113-3120.
[6] Fernandes-Santos C, Carneiro RE, de Souza Mendonca L, et al. Pan-PPAR agonist beneficial effects in overweight mice fed a high-fat high-sucrose diet[J]. Nutrition,2009, 25(7-8):818-827.
[7] Harms M, Seale P.Brown and beige fat: development, function and therapeutic potential[J]. Nat Med,2013,19(10):1252-1263.
[8] Price NL, Fernándezhernando C .miRNA regulation of white and brown adipose tissue differentiation and function[J]. Biochim Biophys Acta, 2016,1861(12PtB):2104-2110.
[9] Kim SY, Kim AY, Lee HW,et al.miR-27a is a negative regulator of adipocyte differentiation via suppressing PPARgamma expression[J]. Biochem Biophys Res Commun,2010,392(3):323-328.
[10] Karbiener M, Fischer C, Nowitsch S, et al.microRNA miR-27b impairs human adipocyte differentiation and targets PPARgamma[J]. Biochem Biophys Res Commun,2009,390(2):247-251.
[11] Lin Q, Gao Z, Alarcon RM, et al.A role of miR-27 in the regulation of adipogenesis[J]. FEBS J,2009,276(8):2348-2358.
[12] Lee EK, Lee MJ, Abdelmohsen K, et al.miR-130 suppresses adipogenesis by inhibiting peroxisome proliferator-activated receptor gamma expression[J]. Mol Cell Biol,2011,31(4):626-638.
[13] Kim YJ, Hwang SJ, Bae YC, et al.MiR-21 regulates adipogenic differentiation through the modulation of TGF-beta signaling in mesenchymal stem cells derived from human adipose tissue[J]. Stem Cells, 2009,27(12):3093-3102.
[14] Yi C, Xie WD, Li F, et al.MiR-143 enhances adipogenic differentiation of 3T3-L1 cells through targeting the co-ding region of mouse pleiotrophin[J]. FEBS Lett,2011, 585(20):3303-3309.
[15] Villarreal-Molina MT, Antuna-Puente B.Adiponectin: anti-inflammatory and cardioprotective effects[J]. Biochimie,2012,94(10):2143-2149.
[16] Guerre-Millo M.Adiponectin: an update[J]. Diabetes Metab,2008,34(1):12-18.
[17] Awazawa M, Ueki K, Inabe K, et al.Adiponectin enhances insulin sensitivity by increasing hepatic IRS-2 expression via a macrophage-derived IL-6-dependent pathway[J]. Cell Metab,2011,13(4):401-412.
[18] Kadowaki T, Yamauchi T, Kubota N, et al.Adiponectin and adiponectin receptors in insulin resistance, diabetes, and the metabolic syndrome[J]. J Clin Invest,2006,116(7):1784-1792.
[19] Frühbeck G, Catalán V, Rodríguez A, et al.Adiponectin-leptin ratio: A promising index to estimate adipose tissue dysfunction. Relation with obesity-associated cardiometabolic risk[J]. Adipocyte,2017:1-6.
[20] Bhaumik D, Scott GK, Schokrpur S, et al.Expression of microRNA-146 suppresses NF-kappaB activity with reduction of metastatic potential in breast cancer cells[J]. Oncogene,2008,27(42):5643-5647.
[21] Hulsmans M, van Dooren E, Mathieu C, et al. Decrease of miR-146b-5p in monocytes during obesity is associa-ted with loss of the anti-inflammatory but not insulin signaling action of adiponectin[J]. PLoS One,2012,7(2):e32794.