摘要
目的通过检测3T3-L1脂肪细胞内糖酵解关键酶的表达情况,探讨课题组前期制备的脂联素球状结构域(gAd)对3T3-L1脂肪细胞葡萄糖分解代谢的影响。方法用课题组前期制备的gAd(质量浓度依次为10、50、100、300、1 000 ng/mL)干预分化成熟的3T3-L1脂肪细胞,干预结束后以实时荧光定量聚合酶链反应(RT-PCR)法检测各组细胞糖酵解关键酶包括己糖激酶、6-磷酸果糖激酶-1及丙酮酸激酶转录表达情况,以Western blot法检测各组细胞丙酮酸激酶翻译表达情况。结果gAd干预组(5个浓度)己糖激酶的转录表达[(2.02±0.16)、(3.47±0.29)、(4.22±0.33)、(5.83±0.45)、(6.65±0.51)倍]显著高于对照组(1.00±0.00)(F=125.789,P<0.001),6-磷酸果糖激酶-1和丙酮酸激酶的转录表达也显著高于对照组(F分别为85.399和113.661,P均<0.001),同时丙酮酸激酶的翻译表达(12.430%±0.800%、1.700%±3.010%、26.570%±1.114%、52.600%±2.910%、71.130%±10.600%)也显著高于对照组(8.000%±1.610%)(F=161.007,P<0.01)。结论 gAd促进3T3-L1脂肪细胞摄取胞外葡萄糖的同时,激活了细胞内葡萄糖的分解供能代谢途径。
Objective To investigate the effect of globular domain of adiponectin（gAd） on glycolysis in 3T3-L1 adipocytes by detecting expression of the key enzymes in the process of glycolysis.Methods Matured 3T3-L1 adipocytes were intervened with pre-prepared gAd（mass concentrations of 10,50,100,300,and 1 000 ng/mL,respectively） and the expression of several key enzymes for glycolysis such as hexokinase,6-phosphofructokinase 1 and pyruvate kinase were determined by real-time polymerase chain reaction（RT-PCR）.Pyruvate kinase protein level was also determined by Western blot.Results The level of hexokinase mRNA in the gAd intervention group was significantly higher than those of the control group[（2.02 ± 0.16）,（3.47 ± 0.29）,（4.22 ± 0.33）,（5.83 ± 0.45） and（6.65 ± 0.51） times of the control]（F = 125.789,P 0.001）.The mRNA levels of 6-phosphofructokinase-1 and pyruvate kinase were also significantly higher than those of the control group（F = 85.399 and F = 113.661,P 0.001）.Consistent with the mRNA expression,the level of pyruvate kinase protein（12.430 % ± 0.800 %,1.700 % ± 3.010 %,26.570 % ± 1.114 %,52.600 % ± 2.910 %,71.130 % ± 10.600 %） was significantly higher than those of the control group（8.000 % ± 1.610 %）（F = 161.007,P 0.01）.Conclusion It is demonstrated that gAd could promote 3T3-L1 cells in taking in glucose from outside of the cells,and in the meantime,activate the glycolysis pathway in 3T3-L1 adipocytes.
出处
《生物医学工程与临床》
CAS
2013年第1期77-81,共5页
Biomedical Engineering and Clinical Medicine
基金
山西省科技攻关项目(20090321104)
山西医科大学第一医院院基金
