利用不同方法将小分子NADPH 转染进入细胞的比较

doi:10.16352/j.issn.1001-6325.2024.05.0613

基础医学与临床 ›› 2024, Vol. 44 ›› Issue (5): 613-618.doi: 10.16352/j.issn.1001-6325.2024.05.0613

利用不同方法将小分子NADPH 转染进入细胞的比较

李皓玥, 杜文静, 李薇^*

中国医学科学院基础医学研究所北京协和医学院基础学院细胞生物学系重大疾病共性机制研究全国重点实验室,北京 100005

收稿日期:2024-01-16 修回日期:2024-03-19 出版日期:2024-05-05 发布日期:2024-04-23
通讯作者: ^*liwei@ibms.pumc.edu.cn
基金资助:
国家重点研发计划(2022YFA0806302,2019YFA0802600);细胞生态海河实验室项目(22HHXBSS0011);中国医学科学院医学与健康科技创新工程(2021-I2M-1-016)

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

LI Haoyue, DU Wenjing, LI Wei^*

Department of Cell Biology, State Key Laboratory of Common Mechanism Research for Major Diseases, Institute of Basic Medical Sciences CAMS, School of Basic Medicine PUMC, Beijing 100005, China

Received:2024-01-16 Revised:2024-03-19 Online:2024-05-05 Published:2024-04-23
Contact: ^*liwei@ibms.pumc.edu.cn

摘要/Abstract

摘要： 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(10 μmol/L NADPH与10 μL 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。

关键词: 酰胺腺嘌呤二核苷酸磷酸(NADPH), 转染, 细胞培养, 脂肪细胞分化

Abstract: 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 (10 μmol/L NADPH and 10 μL 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.

Key words: nicotinamide adenine dinucleotide phosphate(NADPH), transfection, cell culture, adipocyte differentiation

中图分类号:

李皓玥, 杜文静, 李薇. 利用不同方法将小分子NADPH 转染进入细胞的比较[J]. 基础医学与临床, 2024, 44(5): 613-618.

LI Haoyue, DU Wenjing, LI Wei. Application of different methods used to transfect small molecule NADPH into cells[J]. Basic & Clinical Medicine, 2024, 44(5): 613-618.

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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

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