血清剥夺对原代培养大鼠脂肪细胞脂代谢相关基因转录表达的影响  

Effect of Serum-Deprivation on Transcription Expression of Lipid Metabolic-Related Gene in Primary Cultured Rat Adipocytes

作　　者：卢建雄[1] 杨公社[1] 陈粉粉[1] 

机构地区：[1]西北农林科技大学动物脂肪沉积和肌肉发育实验室

出　　处：《畜牧兽医学报》2007年第1期46-52,共7页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家重点基础研究发展计划"973"项目(2004CB117506);国家自然科学基金项目(30471239)

摘　　要：利用血清剥夺/血清培养实验模型,采用油红O染色提取法、甘油试剂盒和RT-PCR,诱导和测定了大鼠附睾脂肪垫分离和培养的脂肪细胞生脂和脂解及其相关酶和转录因子mRNA水平的变化,探讨脂肪细胞脂代谢通路中关键酶和转录因子基因转录表达的规律。结果显示,依血清剥夺时间延长,脂肪细胞激素敏感脂酶(HSL)mRNA表达水平显著提高(P<0.05),恢复血清培养后,表达水平显著降低(P<0.05),与脂解活性的变化一致;脂肪生成及生脂相关基因脂肪酸合酶(FAS)、乙酰辅酶A羧化酶(ACC1)和碳水化合物反应元件结合蛋白(ChREBP)mRNA表达水平依血清剥夺持续时间明显下降(P<0.05),恢复血清培养,随培养时间延长显著提高(P<0.05);血清剥夺显著降低固醇调控元件结合蛋白(SREBP)-1c mRNA水平,但血清剥夺72 h与36 h无明显差异(P>0.05);血清剥夺72 h后恢复血清培养36 h和72 h,SREBP-1c mRNA的表达水平没有显著性变化(P>0.05)。结果表明,生脂及FAS和ACC1表达与ChREBP mRNA水平一致,但与SREBP-1c不一致,提示ChREBP与成熟脂肪细胞生脂基因转录激活可能有更密切的关系,但尚待在蛋白水平进一步证实。Rat preadipocytes were isolated by collagenase treatment of epididymal adipose tissue from male SD rats （20-day-old）, and differentiated into adipocytes. After induced with experiment model of serum-deprivation/re-serum culture, the lipogenesis, lipolysis and relative levels of lipid metabolic-related gene mRNAs in mature adipocytes were measured through Oil Red O staining, glycerol reagent kit and semiquantitative RT-PCR, respectively. The results showed that mRNA levels of hormone-sensitive lipase （HSL） were increased according to lasting time of serum-deprivation （P〈 0. 05）, and significantly decreased with re-serum culture （P 〈 0. 05）, which coincided with the changes of lipolytic activity in adipocytes. Lipogenesis and mRNA levels of lipogenic genes such as fatty acid synthase （FAS）, acetyl-CoA carboxylase α（ACC1） and carbohydrate response element binding protein （ChREBP） were coordinately reduced according to lasting time of serum-deprivation （P〈0.05）, and increasingly increased during re-serum medium culture （P〈0.05）. Sterol regulatory element binding protein （SREBP）-1c mRNA levels were reduced by serum-deprivation （P〈0.05）, but did not have significant difference between 36 h and 72 h of serum-deprivation （P〈0.05）. SREBP-1c mRNA levels were not changed by re-serum medium culture 36 -72 h after 72 h serum-deprivation （P〉0.05）. It can be concluded that lipogenesis and lipogenic genes mRNA levels coincide with mRNA levels of ChREBP, but do not with that of SREBP-1c, suggesting that ChREBP could be relative to transactivation of FAS and ACC1 in adipocytes, which needs to study further in translation level.

关 键 词：脂肪细胞 脂肪生成 脂解 转录 

分 类 号：Q753[生物学—分子生物学]