摘要
为研究溶酶体组织蛋白酶B(cathepsin B,CTSB)对脂肪细胞分化的影响,本实验构建了Ctsb重组腺病毒超表达载体,包装并侵染体外培养的猪前体脂肪细胞,采用油红O染色,油红O提取比色法检测猪前体脂肪细胞分化的情况,并通过real-time PCR法检测成脂关键基因mRNA水平的变化.结果显示,重组腺病毒Ctsb载体构建成功,转染猪前体脂肪细胞后,使Ctsb的mRNA和蛋白质表达量分别提高了约16倍和12倍.CTSB超表达能促进脂肪细胞的分化和脂质积累,成脂关键基因过氧化物酶体增殖物激活受体γ(peroxisome proliferator-activated receptor gamma,PPARγ)、脂肪酸结合蛋白2(adipocyte protein 2,aP2)的表达量均有显著升高.研究表明,提高Ctsb的表达能促进猪前体脂肪细胞分化,揭示了Ctsb在猪前体脂肪细胞分化过程中可能发挥关键作用.研究结果为进一步研究其作用机制奠定了基础.
To investigate the function of lysosomal cathepsin B （CTSB） on adipocyte differentiation, weconstructed a recombinant retroviral vector of Ctsb and used to infect the porcine preadipocytes. Oil red Ostaining and oil red O extraction were used to detect the degree of preadipocyte differentiation. ThemRNA and protein expression of key adipogenic genes were examined by real-time PCR and Westernblotting. The overexpression of CTSB as observed by 12 - 16 folds promoted adipocyte differentiation andlipid accumulation. The expression of peroxisome proliferator-activated receptor gamma （PPARγ） andadipocyte protein2 （ap2） were markedly increased. The results indicated that Ctsb overexpressionfacilitated porcine preadipocyte differentiation.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2014年第7期678-684,共7页
Chinese Journal of Biochemistry and Molecular Biology
基金
高繁殖力猪转基因新品种培育(No.2013ZX08006-005)
国家生猪产业技术体系(No.CARS-36-04)
国家自然科学基金(No.31101685)资助~~
