外源性添加磷酸二羟基丙酮对小鼠脂肪细胞三酰甘油及关键代谢酶含量的影响

Effects of Exogenously Added Dihydroxyacetone Phosphate on the Contents of Triglycerides and Key Metabolic Enzymes in Mouse Adipocytes

旨在研究磷酸二羟基丙酮对小鼠脂肪细胞三酰甘油及其关键代谢酶的影响。将小鼠3T3-L1前脂细胞诱导成成熟的脂肪细胞,在诱导培养第14天,分别加入含0μmol·L^(-1)(ck组)、20μmol·L^(-1)(试验Ⅰ组)、50μmol·L^(-1)(试验Ⅱ组)、200μmol·L^(-1)(试验Ⅲ组)磷酸二羟基丙酮的完全培养基培养细胞,于0、36、72h收集细胞并进行ELISA及后续检测;利用双抗体夹心法检测三酰甘油(TG)代谢中关键酶含量,如乙酰辅酶A羧化酶(ACC)、脂肪酸合成酶(FAS)、激素敏感脂肪酶(HSL)等。结果表明:1)在36和72h,试验Ⅰ、Ⅱ、Ⅲ组合成的TG量极显著高于对照组(ck组)(P<0.01);试验各组葡萄糖-6-磷酸脱氢酶(G6PD)、二硫辛酰胺转乙酰基酶(DLAT)、二硫辛酸脱氢酶(DLD)和脂肪酸合成酶(FAS)含量均极显著高于对照组(P<0.01);2)在36和72h,试验各组激素敏感脂肪酶(HSL)、肉碱脂酰转移酶1(CPT1)和游离脂肪酸(FFA)含量均极显著低于对照组(P<0.01)。磷酸二羟基丙酮能够促进小鼠脂肪细胞三酰甘油的合成和沉积;磷酸二羟基丙酮还能极显著提高TG合成代谢中G6PD、DLAT、DLD、FAS的含量;能够极显著降低TG分解代谢中HSL、CPT1的含量。

The purpose of this study was to investigate the effect of exogenously added dihydroxyacetone phosphate on the contents of triglyceride and its key metabolic enzymes in mouse adipocytes.The inoculated mouse 3 T3-L1 cell into mature adipocytes were the research object,the cells were cultured in dihydroxyacetone phosphate complete medium containing 0μmol·L^-1（ck group）,20μmol·L^-1（group Ⅰ）,50μmol·L^-1（group Ⅱ）and 200μmol·L^-1（group Ⅲ）on the 14 th day,and the cells were collected at 0,36 and 72 hfor the detection of ELISA and other substances.The contents of key enzymes in triglyceride metabolism were detected by using double antibody sandwich technique,such as acetyl-CoA carboxylase（ACC）,fatty acid synthase（FAS）,hormone-sensitive lipase（HSL）and other enzymes.The results showed that：1）The contents of triglyceride in the group Ⅰ,Ⅱ andⅢ were significantly higher than that in ck group（P〈0.01）at 36 and 72 h;The contents of glucose-6-phosphate dehydrogenase（G6 PD）,dihydrolipoyl transacetylase（DLAT）,dihydrolipoyl dehydrogenase（DLD）and FAS in the group Ⅰ,Ⅱ and Ⅲwere significantly higher than that in ck group（P〈0.01）at 36 and 72 h;2）The contents of HSL,carnitine acyltransferase1（CPT1）and free fatty acid（FFA）in the group Ⅰ,Ⅱ and Ⅲwere significantly lower than that in ck group（P〈0.01）at 36 and 72 h.The dihydroxyacetone phosphate can promote the synthesis and deposition of triglyceride in mouse adipocytes;The dihydroxyacetone phosphate can also significantly increase the contents of G6 PD,DLAT,DLD and FAS in triglyceride synthesis,and can significantly reduce the contents of HSL and CPT1 in triglycerides catabolism.