| Orginal Article |
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| The Effect and Mechanism of Down-Regulated Fsp27 Gene Combined with Myricetin on Lipolysis of 3T3-L1 Adipocytes |
DONG Wei-peng,ZHANG Shao-hua,XU Xiang,YAN Jiong( ) |
| School of Public Health, Shanxi Medical University, Taiyuan 030001, China |
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Abstract Objective: Using RNAi technology to reduce Fsp27 expression combined with Myric and observe the effect of lipid metabolism on 3T3-L1 cells, and explore the regulation mechanism of lipid droplets, development and changes.Methods: 3T3-L1 preadipocytes were routinely cultured and induced differentiation by the hormonal cocktail. The shRNA recombinant vectors were transfected into the differentiated 3T3-L1 adipocytes with lipofectamine. The medium was treated with Myric concentration of 100 μmol/L for 72 h. The oil red O staining photographed the morphology and sizes of lipid droplets. The content level of adipocytes cellular TG and glycerol were tested through enzymatic method to evaluate the lipolysis. Western blot measured the expression levels of Fsp27, the adipose triglyceride lipase (ATGL), hormone sensitive lipase (HSL), p38MAPK, p-p38MAPK and PPARγ.Results: 1. After differentiation of 3T3-L1 cells, the morphology changed from fiber-like to round, accompanied by an increase in cell volume. 2. After combined intervention, compared to the control group, content of adipocytes cellular TG decreased with the increased glycerol in sh-Fsp27 group and Myric group (P＜0.05). Meanwhile, compared to the other three groups, the TG content in Myric+sh-Fsp27 group was lower, and the glycerol content increased (P＜0.05). 3. After combined intervention, compared to the control group, the expression levels of Fsp27 in the other three groups decreased significantly, and the expression levels of ATGL and PPARγ increased significantly (P＜0.05). In addition, the expression levels of HSL and p-p38MAPK/p38MAPK in Myric+sh-Fsp27 group and Myric group was higher than that in sh-Fsp27 group and control group (P＜0.05).Conclusions: 1. The combination of Fsp27 gene silencing and Myric could more obviously improve the efficiency of lipolysis than individual. 2. The regulating effect of Myric on lipolysis of 3T3-L1 adipocytes might be exerted through the MAPK pathway by up-regulating the level of ATGL and HSL. The sh-Fsp27 vectors could promote lipolysis by blocking Fsp27 gene and increasing the expression level of ATGL.
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Received: 09 August 2018
Published: 10 January 2019
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Corresponding Authors:
Jiong YAN
E-mail: 13593131989@126.com
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| [1] |
Esser N, Legrand-Poels S, Piette J , et al. Inflammation as a link between obesity, metabolic syndrome and type 2 diabetes. Diabetes Res Clin Pract, 2014,105(2):141-150.
|
|
|
| [2] |
蒲静, 张雪琳, 刘平生 . 脂滴与线粒体相互作用的研究进展. 生物物理学报, 2012,28(02):83-92.
|
|
|
| [2] |
Pu J, Zhang X L, Liu P S . Progress in studies on the interaction between lipid droplets and mitochondria. Biophysics Reports, 2012,28(02):83-92.
|
|
|
| [3] |
Allott E H, Hursting S D . Obesity and cancer: Mechanistic insights from transdisciplinary studies. Endocr Relat Cancer, 2015,22(6):R365-R386.
|
|
|
| [4] |
于进海, 周林康, 吴丽贞 , 等. 肥胖的基础研究. 生命科学, 2015,27(3):268-279.
|
|
|
| [4] |
Yu J H, Zhou L K, Wu L Z , et al. Basic research in obesity. Chinese Bulletin of Life Sciences, 2015,27(3):268-279.
|
|
|
| [5] |
Matsusue K . A physiological role for fat specific protein 27/cell death-inducing DFF45-like effector C in adipose and liver. Biol Pharm Bull, 2010,33(3):346-350.
|
|
|
| [6] |
王谦 . 沙棘籽渣中五种黄酮类化合物对脂代谢的调控及机理探究. 上海: 华东师范大学, 2014.
|
|
|
| [6] |
Wang Q . Effects of five flavonoid compounds from seed residues of hippophae rhamnoides L. on lipid metabolism and their mechanisms. Shanghai: East China Normal University, 2014.
|
|
|
| [7] |
Wang S, Moustaid-Moussa N, Chen L , et al. Novel insights of dietary polyphenols and obesity. J Nutr Biochem, 2014,25(1):1-18.
|
|
|
| [8] |
Wang Q, Wang S T, Yang X , et al. Myricetin suppresses differentiation of 3 T3-L1 preadipocytes and enhances lipolysis in adipocytes. Nutr Res, 2015,35(4):317-327.
|
|
|
| [9] |
Ambesajir A, Kaushik A, Kaushik J J , et al. RNA interference: A futuristic tool and its therapeutic applications. Saudi J Biol Sci, 2012,19(4):395-403.
|
|
|
| [10] |
师磊 . 沉默MCHR1对小鼠3T3-L1细胞脂肪合成的影响及机制研究. 山西: 山西医科大学, 2014.
|
|
|
| [10] |
Shi L . The effect and mechanism of MCHR1 silencing on fat synthesis of mouse 3T3-L1 cells. Shanxi: Shanxi Medical University, 2014.
|
|
|
| [11] |
王君实 . 沉默PLIN1基因与杨梅素联合作用对3T3-L1细胞脂解的影响及机制探究. 山西: 山西医科大学, 2017.
|
|
|
| [11] |
Wang J S . The combined effect and mechanism of PLIN1 gene silencing and myricetin on lipolysis of 3T3-L1 adipocytes. Shanxi: Shanxi Medical University, 2017.
|
|
|
| [12] |
Karbowska J, Kochan Z . Intermittent fasting up-regulates Fsp27/Cidec gene expression in white adipose tissue. Nutrition, 2012,28(3):294-299.
|
|
|
| [13] |
De La Rosa Rodriguez M A, Kersten S . Regulation of lipid droplet-associated proteins by peroxisome proliferator-activated receptors. Biochim Biophys Acta, 2017,1862(10 Pt B):1212-1220.
|
|
|
| [14] |
Xu X, Park J G, So J S , et al. Transcriptional activation of Fsp27 by the liver-enriched transcription factor CREBH promotes lipid droplet growth and hepatic steatosis. Hepatology, 2015,61(3):857-869.
|
|
|
| [15] |
Flach R J, Qin H, Zhang L , et al. Loss of mitogen-activated protein kinase phosphatase-1 protects from hepatic steatosis by repression of cell death-inducing DNA fragmentation factor A (DFFA)-like effector C (CIDEC)/fat-specific protein 27. J Biol Chem, 2011,286(25):22195-22202.
|
|
|
| [16] |
Xiong Y, Collins Q F, An J , et al. p38 mitogen-activated protein kinase plays an inhibitory role in hepatic lipogenesis. J Biol Chem, 2007,282(7):4975-4982.
|
|
|
| [17] |
Lampidonis A D, Rogdakis E, Voutsinas G E , et al. The resurgence of Hormone-Sensitive Lipase (HSL) in mammalian lipolysis. Gene, 2011,477(1-2):1-11.
|
|
|
| [18] |
Girousse A, Langin D . Adipocyte lipases and lipid droplet-associated proteins: insight from transgenic mouse models. Int J Obes (Lond), 2012,36(4):581-594.
|
|
|
| [19] |
Singh M, Kaur R, Lee M J , et al. Fat-specific protein 27 inhibits lipolysis by facilitating the inhibitory effect of transcription factor Egr1 on transcription of adipose triglyceride lipase. J Biol Chem, 2014,289(21):14481-14487.
|
|
|
| [20] |
Yang X, Heckmann B L, Zhang X , et al. Distinct mechanisms regulate ATGL-mediated adipocyte lipolysis by lipid droplet coat proteins. Mol Endocrinol, 2013,27(1):116-126.
|
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