为探讨腺病毒36 型在人脂肪细胞分化过程中对PPARγ及CIDEC 基因表达的调节作用,利用腺病毒感染人脂肪源性间充质干细胞(hAMSC)、油红O 染色和RT-qPCR 鉴定Ad36 诱导hAMSC 分化为脂肪细胞模型;葡萄糖氧化酶法和甘油三酯终点法测定人类腺病毒36 型(Ad36)诱导hAMSC 分化为脂肪细胞的过程中培养基葡萄糖浓度及细胞甘油三酯含量;RT-qPCR、Western Blotting 方法检测Ad36 诱导的人脂肪细胞中,PPARγ和CIDEC 蛋白表达水平的变化;用PPARγ特异性抑制剂GW9662 抑制PPARγ表达后,Western Blotting 方法检测Ad36 诱导的人脂肪细胞中CIDEC 蛋白质的表达。Ad36 诱导的hAM-SC 定向分化成人脂肪细胞,分化过程中培养基葡萄糖含量较对照组显著降低(P<0.05),细胞内甘油三酯含量较对照组显著升高(P<0.05),PPARγ和CIDEC 基因表达水平较对照组显著升高(P<0.05),在诱导第6 天表达水平最高,在使用GW9662 抑制PPARγ蛋白质表达后,CIDEC 蛋白质表达水平较对照组显著降低(P<0.05)。从细胞水平证实,Ad36 诱导人脂肪细胞分化过程中,Ad36 通过PPARγ上调CIDEC 基因的表达水平。
[1] 张春雨, 王德生, 张国华, 等. 中国内蒙古自治区牧区蒙、汉民族55岁 以上居民超重和肥胖病患病率的流行病学调查[J]. 中华内分泌代谢 杂志, 2014, 30(6): 464-467. Zhang Chunyu, Wang Desheng, Zhang Guohua, et al. Epidemiological investigation on the prevalence of obesity and overweight in Mongolian and Han residents aged over 55 years in the pastoral area of Inner Mongolia, China[J]. Chinese Journal of Endocrinology and Metabolism, 2014, 30(6): 464-467.
[2] Dhurandhar N V, Israel B A, Kolesar J M, et al. Increased adiposity in animals due to a human virus[J]. International Journal of Obesity and Related Metabolic Disorders: Journal of the International Association for the Study of Obesity, 2000, 24(8): 989-996.
[3] Atkinson R L, Dhurandhar N V, Allison D B, et al. Human adenovirus-36 is associated with increased body weight and paradoxical reduction of serum lipids[J]. International Journal of Obesity, 2005, 29(3): 281-286.
[4] Rogers P M, Fusinski K A, Rathod M A, et al. Human adenovirus Ad-36 induces adipogenesis via its E4 orf-1 gene[J]. International Journal of Obesity, 2008, 32(3): 397-406.
[5] Pasarica M, Mashtalir N, Mcallister E J, et al. Adipogenic human adenovirus Ad-36 induces commitment, differentiation, and lipid accumulation in human adipose-derived stem cells[J]. Stem Cells, 2008, 26(4): 969-978.
[6] Wu C, Zhang Y, Sun Z, et al. Molecular evolution of Cide family proteins: novel domain formation in early vertebrates and the subsequent divergence[J]. BMC Evolutionary Biology, 2008, 8: 159-174.
[7] Jiao Y, Mao X, Chang X, et al. Adenovirus36 infection expresses cellular APMI and Visfatin genes in overweight Uygur individuals[J]. Diagnostic Pathology, 2014, 9: 83-89.
[8] 努尔比耶·努尔麦麦提, 焦谊, 伊力亚斯·艾萨, 等. 腺病毒36感染对 人脂肪源性间充质干细胞PPARγ和adiponectin 基因表达的影响[J]. 新 疆医科大学学报, 2015, 38(5): 558-563. Nuerbiye Nuermaimaiti, Jiao Yi, Yiliyasi Aisa, et al. The variation of PPARγ and adiponectin genes expression in adenouirus-36 infected human adipose-derived mesenchymal stem cell[J]. Journal of Xinjiang Medical University, 2015, 38(5): 558-563.
[9] Allison D B, Downey M, Atkinson R L, et al. Obesity as a disease: A white paper on evidence and arguments commissioned by the Council of the Obesity Society[J]. Obesity, 2008, 16(6): 1161-1177.
[10] Yeh W C, Cao Z, Classon M, et al. Cascade regulation of terminal adipocyte differentiation by three members of the C/EBP family of leucine zipper proteins[J]. Genes & Development, 1995, 9(2): 168-181.
[11] Hamm J K, El Jack A K, Pilch P F, et al. Role of PPAR gamma in regulating adipocyte differentiation and insulin-responsive glucose uptake[J]. Annals of the New York Academy of Sciences, 1999, 892 (1): 134-145.
[12] Wu Z, Rosen E D, Brun R, et al. Cross-regulation of C/EBP alpha and PPAR gamma controls the transcriptional pathway of adipogenesis and insulin sensitivity[J]. Molecular Cell, 1999, 3(2): 151-158.
[13] Kawada T. Lipid metabolism related nuclear receptor-The structure, function, expression and classification of peroxisome proliferationactivated receptor (PPAR) [J]. Nihon Rinsho. Japanese Journal of Clinical Medicine, 1998, 56(7): 1722-1728.
[14] Takeyama K, Kodera Y, Suzawa M, et al. Peroxisome proliferatoractivated receptor (PPAR)-structure, function, tissue distribution, gene expression[J]. Nihon Rinsho. Japanese Journal of Clinical Medicine, 2000, 58(2): 357-363.
[15] Fajas L, Fruchart J C, Auwerx J. PPARgamma3 mRNA: A distinct PPARgamma mRNA subtype transcribed from an independent promoter[J]. FEBS Letters, 1998, 438(1-2): 55-60.
[16] Elbrecht A, Chen Y, Cullinan C A, et al. Molecular cloning, expression and characterization of human peroxisome proliferator activated receptors gamma 1 and gamma 2[J]. Biochemical and unications, 1996, 224(2): 431-437.
[17] Ren D, Collingwood T N, Rebar E J, et al. PPARgamma knockdown by engineered transcription factors: Exogenous PPARgamma2 but not PPARgamma1 reactivates adipogenesis [J]. Genes & Development, 2002, 16(1): 27-32.
[18] Li J Z, Li P. Cide proteins and the development of obesity[J]. Novartis Foundation Symposium, 2007, 286(155-159): 196-203.
[19] Boren J, Taskinen M R, Olofsson S O, et al. Ectopic lipid storage and insulin resistance: A harmful relationship[J]. Journal of Internal Medicine, 2013, 274(1): 25-40.
[20] Saraf N, Sharma P K, Mondal S C, et al. Role of PPARg2 transcription factor in thiazolidinedione-induced insulin sensitization[J]. The Journal of Pharmacy and Pharmacology, 2012, 64(2): 161-171.
[21] Seale P, Bjork B, Yang W, et al. PRDM16 controls a brown fat/skeletal muscle switch[J]. Nature, 2008, 454(7207): 961-967.
[22] Puri V, Konda S, Ranjit S, et al. Fat-specific protein 27, a novel lipid droplet protein that enhances triglyceride storage[J]. The Journal of Biological Chemistry, 2007, 282(47): 34213-34218.
[23] Keller P, Petrie J T, De Rose P, et al. Fat-specific protein 27 regulates storage of triacylglycerol[J]. The Journal of Biological Chemistry, 2008,283(21): 14355-14365.
[24] Nian Z, Sun Z, Yu L, et al. Fat-specific protein 27 undergoes ubiquitin-dependent degradation regulated by triacylglycerol synthesis and lipid droplet formation[J]. The Journal of Biological Chemistry, 2010, 285(13): 9604-9615.
[25] Matsusue K, Kusakabe T, Noguchi T, et al. Hepatic steatosis in leptindeficient mice is promoted by the PPARgamma target gene Fsp27 [J]. Cell Metabolism, 2008, 7(4): 302-311.
[26] Kim Y J, Cho S Y, Yun C H, et al. Transcriptional activation of Cidec by PPARgamma2 in adipocyte [J]. Biochemical and Biophysical Rsearch Communications, 2008, 377(1): 297-302.