利用不同方法将小分子NADPH转染进入细胞的比较

Application of different methods used to transfect small molecule NADPH into cells

目的利用非代谢途径直接将外源小分子烟酰胺腺嘌呤二核苷酸磷酸(NADPH)转染进入细胞内的方法。方法对比3种不同转染试剂(X-tremeGENE TM HP DNA、Lipofectamine TM RNAiMAX和Lipofectamine TM 2000)将NADPH转染到人骨肉瘤细胞系U2OS和小鼠胚胎成纤维细胞系3T3L1中的效果,并通过油红O染色比较它们对脂肪细胞分化的影响。结果用X-tremeGENE HP DNA转染试剂转染NADPH可以有效提高细胞内NADPH水平(P<0.001)。随着NADPH转染浓度(10μmol/L NADPH与10μL转染试剂)的增加,细胞中的NADPH水平呈剂量依赖性增加。此外使用3种转染试剂在3T3L1前脂肪细胞中转染NADPH,只有使用X-tremeGENE HP DNA转染试剂转染NADPH的脂肪细胞分化更明显(P<0.001)。结论X-tremeGENE HP DNA转染试剂能够成功地将外源NADPH转染进入细胞内,并促进3T3L1脂肪细胞的分化和脂质积累。

Objective To compare the three different methods for direct transfection of small exogenous nicotinamide adenine dinucleotide phosphate(NADPH)into cells by a non-metabolic pathway.Methods Three different transfection reagents(X-tremeGENE TM HP DNA,Lipofectamine TM RNAiMAX and Lipofectamine TM 2000)were used to find the transfection efficiency of NADPH into human osteosarcoma cell line U2OS and mouse embryonic fibroblast cell line 3T3L1.Their effects on adipocyte differentiation were compared by Oil Red O staining.Results Transfection of NADPH with X-tremeGENE HP DNA transfection reagent effectively increased intracellular NADPH level(P<0.001).With the increase of NADPH transfection concentration(10μmol/L NADPH and 10μL transfection reagent),the level of NADPH in cells increased in a dose-dependent manner.In addition,three transfection reagents were used to transfect NADPH in 3T3L1 preadipocytes.Only cells transfected with NADPH using X-tremeGENE HP DNA transfection reagent contained more lipids than control cells(P<0.001).Conclusions Transfection of NADPH with X-tremeGENE HP DNA transfection reagent effectively increases the level of NADPH in cells.