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,PPARγ、FAS、ACC、AP2和CEBPα的mRNA表达量也极显著上调(P<0.01或P<0.001),ATGL和HSL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。
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 receptorγ(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 CEBPαwere 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 accumulatio
作者
谢红月
潘鹏
胡艳
张瑜
罗云彦
胡旭旭
甄锐
蒋钦杨
黄艳娜
XIE Hongyue;PAN Peng;HU Yan;ZHANG Yu;LUO Yunyan;HU Xuxu;ZHEN Rui;JIANG Qinyang;HUANG Yanna(College of Animal Science and Technology,Guangxi University,Nanning 530004,China)
出处
《动物营养学报》
CAS
CSCD
北大核心
2021年第6期3487-3496,共10页
CHINESE JOURNAL OF ANIMAL NUTRITION
基金
国家自然科学基金项目(31760672,31460606)。
