分子与细胞营养 MOLECULAR AND CELLULAR NUTRITION

高温对3T3-L1前体脂肪细胞凋亡和脂肪沉积的影响

  • 谢红月 ,
  • 潘鹏 ,
  • 胡艳 ,
  • 张瑜 ,
  • 罗云彦 ,
  • 胡旭旭 ,
  • 甄锐 ,
  • 蒋钦杨 ,
  • 黄艳娜
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  • 广西大学动物科学技术学院, 南宁 530004
谢红月(1995-),女,辽宁建平人,硕士研究生,研究方向为动物营养与肉质调控。E-mail:2996970767@qq.com

收稿日期: 2020-11-16

  网络出版日期: 2021-06-10

基金资助

国家自然科学基金项目(31760672,31460606)

Effects of High Temperature on Apoptosis and Fat Deposition of 3T3-L1 Preadipocytes

  • XIE Hongyue ,
  • PAN Peng ,
  • HU Yan ,
  • ZHANG Yu ,
  • LUO Yunyan ,
  • HU Xuxu ,
  • ZHEN Rui ,
  • JIANG Qinyang ,
  • HUANG Yanna
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  • College of Animal Science and Technology, Guangxi University, Nanning 530004, China

Received date: 2020-11-16

  Online published: 2021-06-10

摘要

本试验旨在研究高温对3T3-L1前体脂肪细胞凋亡和脂肪沉积的影响。选择3T3-L1前体脂肪细胞为研究材料,分别以37.0和41.5℃处理细胞,4 d后观察细胞凋亡情况,分化处理6 d后,油红O染色观察脂滴的数量,采用实时荧光定量PCR(qRT-PCR)技术测定热休克蛋白家族基因热休克蛋白60(HSP60)、热休克蛋白70(HSP70)、热休克蛋白90(HSP90),细胞凋亡相关基因半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)和半胱氨酸天冬氨酸蛋白酶-9(Caspase-9)、B细胞淋巴瘤-2(Bcl-2)和Bcl-2相关X蛋白(Bax),脂肪合成相关基因过氧化物酶体增殖剂激活受体γ(PPARγ)、脂肪酸合成酶(FAS)、乙酰辅酶A羧化酶(ACC)、脂肪酸结合蛋白2(AP2)、CCAAT增强子结合蛋α(CEBPα)和脂肪分解相关基因激素敏感脂酶(HSL)、脂肪甘油三酯脂酶(ATGL)的mRNA表达量。结果显示:高温处理后,HSP60、HSP70、HSP90的mRNA表达量极显著上调(P<0.01或P<0.001),同时Caspase-3、Caspase-9 mRNA表达量也极显著上调(P<0.01),而Bcl-2/Bax mRNA比值极显著降低(P<0.001)。细胞内脂滴大量聚集,油红O染色脂滴着色明显,进一步的研究显示,PPARγFASACCAP2和CEBPα的mRNA表达量也极显著上调(P<0.01或P<0.001),ATGLHSL的mRNA表达量极显著下调(P<0.001)。上述结果说明高温会引起3T3-L1前体脂肪细胞的凋亡,促进脂肪酸的合成,抑制脂肪酸的分解,进而增加3T3-L1前体脂肪细胞的脂质累积。

本文引用格式

谢红月 , 潘鹏 , 胡艳 , 张瑜 , 罗云彦 , 胡旭旭 , 甄锐 , 蒋钦杨 , 黄艳娜 . 高温对3T3-L1前体脂肪细胞凋亡和脂肪沉积的影响[J]. 动物营养学报, 2021 , 33(6) : 3487 -3496 . DOI: 10.3969/j.issn.1006-267x.2021.06.050

Abstract

The aim of this study was to investigate the effects of high temperature on the apoptosis and fat deposition of 3T3-L1 preadipocytes. 3T3-L1 preadipocytes were selected as research materials. The cells were treated at 37.0 and 41.5℃ respectively, apoptosis was observed 4 days later, and the number of lipid droplets was observed by oil red O staining after 6 days of differentiation. The mRNA expression levels of heat shock protein family genes involved heat shock protein 60 (HSP60), heat shock protein 70 (HSP70), heat shock protein 90 (HSP90), apoptosis related genes involved cysteine aspartate protease-3 (Caspase-3) and cysteine aspartate protease-9 (Caspase-9), B lymphocytoma-2 (Bcl-2) and Bcl-2-associated X protein (Bax), fat synthesis related genes involved peroxisome proliferators-activated receptor γ (PPARγ), fatty acid synthase (FAS), acetyl CoA carboxylase (ACC), fatty acid-binding protein 2 (AP2), CCAAT/enhancer-binding proteinα (CEBPα), and fat decomposition related genes involved hormone-sensitive triglayceride lipase (HSL) and adipose triglyceride lipase (ATGL) were determined by real-time quantitative PCR (qRT-PCR). The results showed that the mRNA expression levels of HSP60, HSP70 and HSP90 were extremely significantly up-regulated after high temperature treatment (P<0.01 or P<0.001). The mRNA expression levels of Caspase-3 and Caspase-9 were extremely significantly up-regulated (P<0.01), while the Bcl-2/Bax mRNA ratio was extremely significantly decreased (P<0.001). A large number of intracellular lipid droplets were clustered, and the staining of lipid droplets with oil red O staining was obvious. Further studies showed that the mRNA expression levels of PPARγ, FAS, ACC, AP2 and CEBPα were also extremely significantly up-regulated (P<0.01 or P<0.001), while the mRNA expression levels of ATGL and HSL were extremely significantly down-regulated (P<0.001). In conclusion, high temperature can induce apoptosis of 3T3-L1 preadipocytes, promote the synthesis of fatty acids, inhibit the decomposition of fatty acids, and then increase the lipid accumulation of 3T3-L1 preadipocytes.

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