不同茶多糖对3T3-L1前脂肪细胞分化的抑制作用比较

doi:10.7506/spkx1002-6630-201721030

食品科学 ›› 2017, Vol. 38 ›› Issue (21): 187-194.doi: 10.7506/spkx1002-6630-201721030

不同茶多糖对3T3-L1前脂肪细胞分化的抑制作用比较

李娟，刘锐，吴涛，郭郁，张民,

（1.天津科技大学?食品营养与安全教育部重点实验室，天津 300457；2.天津食品安全低碳制造协同创新中心，天津 300457）

出版日期:2017-11-15 发布日期:2017-11-01
基金资助:
“十二五”国家科技支撑计划项目（2012BAD33B08）；天津市应用基础与前沿技术研究计划重点项目（14JCZDJC34800）；天津市应用基础与前沿技术研究计划（青年基金项目）（15JCQNJC14900）

Comparative Study of the Anti-Obesity Effects of Green, Black and Oolong Tea Polysaccharides in 3T3-L1 Preadipocytes

LI Juan, LIU Rui, WU Tao, GUO Yu, ZHANG Min,

(1. Key Laboratory of Food Nutrition and Safety, Ministry of Education, Tianjin University of Science and Technology, Tianjin 300457, China; 2. Tianjin Food Safety and Low Carbon Manufacturing Collaborative Innovation Center, Tianjin 300457, China)

Online:2017-11-15 Published:2017-11-01

摘要/Abstract

摘要： 本实验利用小鼠3T3-L1前脂肪细胞对绿茶多糖（green tea polysaccharides，GTP）、红茶多糖（black tea polysaccharides，BTP）和乌龙茶多糖（oolong tea polysaccharides，OTP）的减肥作用进行评价。使用气相色谱对GTP、BTP、OTP的单糖组成进行分析。采用噻唑蓝染色法测定3?种茶多糖（tea polysaccharides，TPs）对3T3-L1前脂肪细胞增殖活力的影响。使用流式细胞仪检测3?种TPs对3T3-L1前脂肪细胞细胞周期的影响。采用传统“鸡尾酒”法诱导3T3-L1细胞分化成脂后，测吸光度并计算其分化率。采用甘油磷酸氧化酶-过氧化物酶（glycerol phosphate oxidase-peroxidase，GPO-PAP）法测定细胞中甘油三酯（triglyceride，TG）含量的变化。采用实时荧光定量聚合酶链式反应（real-time polymerase chain reaction，RT-PCR）技术测定与脂质代谢相关基因的表达量。结果显示3?种TPs均能显著抑制3T3-L1前脂肪细胞的增殖与分化（P＜0.05）。添加100?μg/mL的TPs能使细胞分化率显著下降至（62.00±6.61）%（GTP）、（82.95±4.25）%（BTP）、（97.24±5.80）%（OTP）。另外，在细胞培养至第5天检测表明，这3 种TPs显著促进G0/G1期细胞数量的累积，细胞比例分别为（68.52±2.28）%（GTP）、（67.11±1.68）%（BTP）、（59.69±1.35）%（OTP）。RT-PCR分析结果显示TPs可以调控相关脂肪细胞因子的表达。在本研究中，在3T3-L1前脂肪细胞中GTP的减肥作用强于BTP和OTP。TPs的加入上调脂联素的表达从而激活腺苷酸活化蛋白激酶信号通路调控相关脂肪细胞因子的表达，并最终抑制TG的合成与3T3-L1前脂肪细胞的分化。

关键词: 茶多糖, 减肥作用, 3T3-L1前脂肪细胞, 分化, 基因表达

Abstract: This study aimed to evaluate the anti-obesity effects of green tea polysaccharides (GTP), black tea polysaccharides (BTP) and oolong tea polysaccharides (OTP) in 3T3-L1 preadipocytes. The monosaccharide compositions of GTP, BTP and OTP were analyzed using gas chromatography, and the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay was used to determine the effects of three tea polysaccharides (TPs) on the proliferation of 3T3-L1 preadipocytes. The cell cycle was determined by flow cytometry. The preadipocytes were differentiated into adipocytes using the traditional “cocktail” method and the differentiation rate was measured based on absorbance. The triglyceride (TG) content was measured to analyze lipid accumulation in adipocytes using the phosphoric acid oxidase peroxidase (GPO-PAP) method. The expression of transcription factors related to adipocyte differentiation was analyzed by real-time polymerase chain reaction (RT-PCR). The inhibition of the proliferation and differentiation of 3T3-L1 preadipocytes by three TPs were significantly different (P < 0.05). With the addition of 100 μg/mL TPs, the differentiation rates were (62.00 ± 6.61)% (GTP), (82.95 ± 4.25)% (BTP) and (97.24 ± 5.80)% (OTP). In addition, GTP, BTP and OTP could regulate the cell cycle, and the proportions of G0/G1 phase cells after 5 days were (68.52 ± 2.28)%, (67.11 ± 1.68)% and (59.69 ± 1.35)%, respectively. Further investigation of the total RNA extracted from the differentiated preadipocytes revealed that the addition of TPs (100 μg/mL) could regulate the expression of related cytokines. In this study, the inhibitory effect of GTP was more significant than those of BTP and OTP. Up-regulation of of adiponectin (ADP) expression activated the adenosine monophosphate-activated protein kinase (AMPK) signaling pathway, which regulates the expression of related adipocyte factors, and ultimately leading to inhibition of TG synthesis and 3T3-L1 preadipocyte differentiation.

Key words: tea polysaccharides (TPs), anti-obesity activity, 3T3-L1 preadipocytes, differentiation, gene expression

中图分类号:

TS201.4

李娟，刘锐，吴涛，郭郁，张民,. 不同茶多糖对3T3-L1前脂肪细胞分化的抑制作用比较[J]. 食品科学, 2017, 38(21): 187-194.

LI Juan, LIU Rui, WU Tao, GUO Yu, ZHANG Min,. Comparative Study of the Anti-Obesity Effects of Green, Black and Oolong Tea Polysaccharides in 3T3-L1 Preadipocytes[J]. FOOD SCIENCE, 2017, 38(21): 187-194.

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不同茶多糖对3T3-L1前脂肪细胞分化的抑制作用比较

Comparative Study of the Anti-Obesity Effects of Green, Black and Oolong Tea Polysaccharides in 3T3-L1 Preadipocytes

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