论文部分内容阅读
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 preadipted adiponectin globulin (gAd) on glucose catabolism in 3T3-L1 adipocytes by detecting the expression of key glycolytic enzymes in 3T3-L1 adipocytes. Methods The 3T3-L1 adipocytes differentiated from mature 3T3-L1 adipocytes were pretreated with gAd (10,50,100,300 and 1, 000 ng / mL, respectively) prepared in the previous study. After the intervention, real-time fluorescent quantitative polymerase chain reaction The expression of key enzymes of glycolysis, including hexokinase, 6-phosphofructokinase-1 and pyruvate kinase, in each group was detected by -PCR method. The translational expression of pyruvate kinase in each group was detected by Western blot. Results The transcriptional expression of hexokinase in the gAd-treated group (5 concentrations) was significantly higher than that of hexokinase [(2.02 ± 0.16), (3.47 ± 0.29), (4.22 ± 0.33), (5.83 ± 0.45), (6.65 ± 0.51) The transcripts of 6-phosphofructokinase-1 and pyruvate kinase were also significantly higher in the control group (F = 125.789, P <0.001) (F = 85.399 and 113.661, respectively, P <0.001) , While translational expression of pyruvate kinase (12.430% ± 0.800%, 1.700% ± 3.010%, 26.570% ± 1.114%, 52.600% ± 2.910%, 71.130% ± 10.600%) was also significantly higher than that of the control group (8.000% ± 1.610 %) (F = 161.007, P <0.01). Conclusions gAd promotes the uptake of extracellular glucose by 3T3-L1 adipocytes and activates the breakdown of intracellular glucose for metabolic pathway.