INFLUENCE OF BURN SERUM ON PROLIFERATION AND ADIPOSE DIFFERENTIATION OF 3T3-L1 PREADIPOCYTES_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

ZHUANGShubo ^1,2 ,  CHAIJiake ¹ , YUYonghui ¹ , DUANHongjie ¹ , LIULingying ¹ , FANJun ¹ , HOUYusen ² , WANGYihe ²

1. Department of Burn and Plastic Surgery, the First Affiliated Hospital, General Hospital of Chinese PLA, Beijing, 100048, P. R. China;
2. Chinese PLA Medical College;

Corresponding author：

CHAIJiake, Email: cjk304@126.com

Keywords：

Burn serum; 3T3-Ll preadipocytes; Adipose differentiation; Rat

DOI：

10.7507/1002-1892.20140169

Video：

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Objective To investigate the effect of burn on the fat metabolism by observing the effect of burn serum on the proliferation and adipose differentiation of 3T3-L1 preadipocytes. Methods Forty-eight male Sprague Dawley rats were randomly divided into sham burn group and burn at 1, 4, 7, 14, and 21 days groups, 8 rats in each group. The rats in burn groups were made the full-thickness thermal burns comprising 30% total body surface area. At 1, 4, 7, 14, and 21 days after burn, the serum of burn rats was collected. The rats in sham burn group were not treated as normal control. The proliferation activity of 3T3-Ll cells was detected using MTT method after treated by normal and burn serum. The burn serum having the highest proliferation inhibitory effect was chosen for subsequent study. The growth of 3T3-L1 cells in normal serum group (group A), burn serum group (group B), normal serum and adipogenic induction group (group C), burn serum and adipogenic induction group (group D) was observed using inverted microscope. After 7 days of treatment, the adipocytes was stained by oil red O and the absorbance (A) value was measured. The mRNA and protein levels of preoxisome proliferator-activated receptor γ (PPAR-γ) and lipoprotein lipase (LPL) were detected by real-time quantitative PCR and Western blot. Results The proliferation ability of 3T3-L1 cells was significantly reduced in the group treated by 4-or 7-day burn serum (P<0.05), especially 7-day burn serum treatment group (P<0.05). Under inverted microscope, the cell morphology in group A and group B had no obvious change, but a large number of fat cells were observed in group C and a few were observed in group D. The positive or weak positive oil red O staining was observed in group C or group D, respectively. The cell counting and A value were significantly higher in group A than in group B, and in group C than in group D (P<0.05). The mRNA level of PPAR-γ in group B was significantly reduced when compared with that in group A (P<0.05). No significant difference was found in LPL mRNA levels and protein levels of PPAR-γ and LPL between group A and group B (P>0.05). The mRNA and protein levels of PPAR-γ and LPL were significantly attenuated in group D when compared with those in group C (P<0.05). Conclusion The adipose differentiation of 3T3-L1 preadipocytes can be significantly reduced after treated by 7-day burn serum of rat.

Citation： ZHUANGShubo, CHAIJiake, YUYonghui, DUANHongjie, LIULingying, FANJun, HOUYusen, WANGYihe. INFLUENCE OF BURN SERUM ON PROLIFERATION AND ADIPOSE DIFFERENTIATION OF 3T3-L1 PREADIPOCYTES. Chinese Journal of Reparative and Reconstructive Surgery, 2014, 28(6): 763-767. doi: 10.7507/1002-1892.20140169 Copy

1.	柴家科, 申传安, 盛志勇. 严重烧伤脓毒症患者骨骼肌蛋白分解代谢的临床研究. 中华医学杂志, 2005, 85(41):2895-2898.
2.	Rabinovici R, Gautschy S, Coimbra R. Trends in trauma surgery:analysis of the American Association for the Surgery of Trauma program 1939-2012. J Trauma Acute Care Surg, 2014, 76(3):672-680.
3.	Carter EA, Paul C, Bonab AA, et al. Effect of exercise on burn-induced changes in tissue-specific glucose metabolism. J Burn Care Res, 2014.[Epub ahead of print].
4.	Heath V. Metabolism:Feel the burn-muscle metabolite couples exercise to heat production. Nat Rev Endocrinol, 2014, 10(4):188.
5.	Rousseau AF, Ledoux D, Damas P, et al. Hypovitaminosis D and burn injury. Rev Med Liege, 2013, 68(11):574-578.
6.	Burmistrova AL, Filippova IuIu. Soluble form of trigger receptor expressed on myeloid cells-1 as a marker of burn wound mixed infection. Zh Mikrobiol Epidemiol Immunobiol, 2013, (6):63-68.
7.	Duan H, Chai J, Sheng Z, et al. Effect of burn injury on apoptosis and expression of apoptosis-related genes/proteins in skeletal muscles of rats. Apoptosis, 2009, 14(1):52-65.
8.	蔡建华, 柴家科, 申传安, 等. 烧伤、冲击伤及烧冲复合伤后大鼠早期血清中性粒细胞弹性蛋白酶的变化及意义. 中华医学杂志, 2010, 90(24):1707-1710.
9.	庄淑波, 刘毅, 李晓东. 大鼠骨髓间充质干细胞与PLGA构建组织工程脂肪的实验研究. 中华损伤与修复杂志(电子版), 2007, 2(5):329-335.
10.	Ruan H, Hacohen N, Golub TR, et al. Tumor necrosis factor-alpha suppresses adipocyte-specific genes and activates expression of preadipocyte genes in 3T3-L1 adipocytes:nuclear factor-kappaB activation by TNF-alpha is obligatory. Diabetes, 2002, 51(5):1319-1336.
11.	Lee MJ, Fried SK. The glucocorticoid receptor, not the mineralocorticoid receptor, plays the dominant role in adipogenesis and adipokine production in human adipocytes. Int J Obes (Lond), 2014.[Epub ahead of print].
12.	Bae CR, Park YK, Cha YS. Quercetin-rich onion peel extract suppresses adipogenesis by down-regulating adipogenic transcription factors and gene expression in 3T3-L1 adipocytes. J Sci Food Agric, 2014.[Epub ahead of print].
13.	Yan H, Kermouni A, Abdel-Hafez M, et al. Role of cyclooxygenases COX-1 and COX-2 in modulating adipogenesis in 3T3-L1 cells. J Lipid Res, 2003, 44(2):424-429.
14.	Cinel I, Opal SM. Molecular biology of inflammation and sepsis:a primer. Crit Care Med, 2009, 37(1):291-304.
15.	Dehne MG, Sablotzki A, Hoffmann A, et al. Alterations of acute phase reaction and cytokine production in patients following severe burn injury. Burns, 2002, 28(6):535-542.
16.	Liao FH, Liou TH, Chiu WC, et al. Differential effects of high MUFA with high or low P/S ratio (polyunsaturated to saturated fatty acids) on improving hepatic lipolytic enzymes and mediating PPARγ related with lipoprotein lipase and hormon-sensitive lipase of white adipose tissue in diet-induced obese hamster. Int J Obes (Lond), 2010, 34(11):1608-1617.
17.	Chen L, Yang G. PPARs Integrate the Mammalian Clock and Energy Metabolism. PPAR Res, 2014, 2014:653017.
18.	Park M, Lee JH, Choi JK, et al. 18β-glycyrrhetinic acid attenuates nandamide-induced adiposity and high-fat diet induced obesity. Mol Nutr Food Res, 2014.[Epub ahead of print].
19.	Bartelt A, Heeren J. Adipose tissue browning and metabolic health. Nat Rev Endocrinol, 2014, 10(1):24-36.
20.	Palou M, Priego T, Romero M, et al. Moderate calorie restriction during gestation programs offspring for lower BAT thermogenic capacity driven by thyroid and sympathetic signaling. Int J Obes (Lond), 2014.[Epub ahead of print].
21.	Daoud MS, Ataya FS, Fouad D, et al. Associations of three lipoprotein lipase gene polymorphisms, lipid profiles and coronary artery disease. Biomed Rep, 2013, 1(4):573-582.
22.	Kim JH, Meyers MS, Khuder SS, et al. Tissue-selective estrogen complexes with bazedoxifene prevent metabolic dysfunction in female mice. Mol Metab, 2014, 3(2):177-190.
23.	Cha SC, Qin H, Sakamaki I, et al. Cloning variable region genes of clonal lymphoma immunoglobulin for generating patient-specific idiotype DNA vaccine. Methods Mol Biol, 2014, 1139:289-303.
24.	Wang Y, Pei-Ling Chiu A, Neumaier K, et al. Endothelial cell heparanase taken up by cardiomyocytes regulates lipoprotein lipase transfer to the coronary lumen following diabetes. Diabetes, 2014.[Epub ahead of print].
25.	Cougnard-Grégoire A, Delyfer MN, Korobelnik JF, et al. Elevated high-density lipoprotein cholesterol and age-related macular degeneration:the Alienor study. PLoS One, 2014, 9(3):e90973.
26.	Bentley AR, Chen G, Shriner D, et al. Gene-based sequencing identifies lipid-influencing variants with ethnicity-specific effects in african americans. PLoS Genet, 2014, 10(3):e1004190.

1. 柴家科, 申传安, 盛志勇. 严重烧伤脓毒症患者骨骼肌蛋白分解代谢的临床研究. 中华医学杂志, 2005, 85(41):2895-2898.
2. Rabinovici R, Gautschy S, Coimbra R. Trends in trauma surgery:analysis of the American Association for the Surgery of Trauma program 1939-2012. J Trauma Acute Care Surg, 2014, 76(3):672-680.
3. Carter EA, Paul C, Bonab AA, et al. Effect of exercise on burn-induced changes in tissue-specific glucose metabolism. J Burn Care Res, 2014.[Epub ahead of print].
4. Heath V. Metabolism:Feel the burn-muscle metabolite couples exercise to heat production. Nat Rev Endocrinol, 2014, 10(4):188.
5. Rousseau AF, Ledoux D, Damas P, et al. Hypovitaminosis D and burn injury. Rev Med Liege, 2013, 68(11):574-578.
6. Burmistrova AL, Filippova IuIu. Soluble form of trigger receptor expressed on myeloid cells-1 as a marker of burn wound mixed infection. Zh Mikrobiol Epidemiol Immunobiol, 2013, (6):63-68.
7. Duan H, Chai J, Sheng Z, et al. Effect of burn injury on apoptosis and expression of apoptosis-related genes/proteins in skeletal muscles of rats. Apoptosis, 2009, 14(1):52-65.
8. 蔡建华, 柴家科, 申传安, 等. 烧伤、冲击伤及烧冲复合伤后大鼠早期血清中性粒细胞弹性蛋白酶的变化及意义. 中华医学杂志, 2010, 90(24):1707-1710.
9. 庄淑波, 刘毅, 李晓东. 大鼠骨髓间充质干细胞与PLGA构建组织工程脂肪的实验研究. 中华损伤与修复杂志(电子版), 2007, 2(5):329-335.
10. Ruan H, Hacohen N, Golub TR, et al. Tumor necrosis factor-alpha suppresses adipocyte-specific genes and activates expression of preadipocyte genes in 3T3-L1 adipocytes:nuclear factor-kappaB activation by TNF-alpha is obligatory. Diabetes, 2002, 51(5):1319-1336.
11. Lee MJ, Fried SK. The glucocorticoid receptor, not the mineralocorticoid receptor, plays the dominant role in adipogenesis and adipokine production in human adipocytes. Int J Obes (Lond), 2014.[Epub ahead of print].
12. Bae CR, Park YK, Cha YS. Quercetin-rich onion peel extract suppresses adipogenesis by down-regulating adipogenic transcription factors and gene expression in 3T3-L1 adipocytes. J Sci Food Agric, 2014.[Epub ahead of print].
13. Yan H, Kermouni A, Abdel-Hafez M, et al. Role of cyclooxygenases COX-1 and COX-2 in modulating adipogenesis in 3T3-L1 cells. J Lipid Res, 2003, 44(2):424-429.
14. Cinel I, Opal SM. Molecular biology of inflammation and sepsis:a primer. Crit Care Med, 2009, 37(1):291-304.
15. Dehne MG, Sablotzki A, Hoffmann A, et al. Alterations of acute phase reaction and cytokine production in patients following severe burn injury. Burns, 2002, 28(6):535-542.
16. Liao FH, Liou TH, Chiu WC, et al. Differential effects of high MUFA with high or low P/S ratio (polyunsaturated to saturated fatty acids) on improving hepatic lipolytic enzymes and mediating PPARγ related with lipoprotein lipase and hormon-sensitive lipase of white adipose tissue in diet-induced obese hamster. Int J Obes (Lond), 2010, 34(11):1608-1617.
17. Chen L, Yang G. PPARs Integrate the Mammalian Clock and Energy Metabolism. PPAR Res, 2014, 2014:653017.
18. Park M, Lee JH, Choi JK, et al. 18β-glycyrrhetinic acid attenuates nandamide-induced adiposity and high-fat diet induced obesity. Mol Nutr Food Res, 2014.[Epub ahead of print].
19. Bartelt A, Heeren J. Adipose tissue browning and metabolic health. Nat Rev Endocrinol, 2014, 10(1):24-36.
20. Palou M, Priego T, Romero M, et al. Moderate calorie restriction during gestation programs offspring for lower BAT thermogenic capacity driven by thyroid and sympathetic signaling. Int J Obes (Lond), 2014.[Epub ahead of print].
21. Daoud MS, Ataya FS, Fouad D, et al. Associations of three lipoprotein lipase gene polymorphisms, lipid profiles and coronary artery disease. Biomed Rep, 2013, 1(4):573-582.
22. Kim JH, Meyers MS, Khuder SS, et al. Tissue-selective estrogen complexes with bazedoxifene prevent metabolic dysfunction in female mice. Mol Metab, 2014, 3(2):177-190.
23. Cha SC, Qin H, Sakamaki I, et al. Cloning variable region genes of clonal lymphoma immunoglobulin for generating patient-specific idiotype DNA vaccine. Methods Mol Biol, 2014, 1139:289-303.
24. Wang Y, Pei-Ling Chiu A, Neumaier K, et al. Endothelial cell heparanase taken up by cardiomyocytes regulates lipoprotein lipase transfer to the coronary lumen following diabetes. Diabetes, 2014.[Epub ahead of print].
25. Cougnard-Grégoire A, Delyfer MN, Korobelnik JF, et al. Elevated high-density lipoprotein cholesterol and age-related macular degeneration:the Alienor study. PLoS One, 2014, 9(3):e90973.
26. Bentley AR, Chen G, Shriner D, et al. Gene-based sequencing identifies lipid-influencing variants with ethnicity-specific effects in african americans. PLoS Genet, 2014, 10(3):e1004190.

Chinese Journal of Reparative and Reconstructive Surgery

INFLUENCE OF BURN SERUM ON PROLIFERATION AND ADIPOSE DIFFERENTIATION OF 3T3-L1 PREADIPOCYTES

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