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学科主题: 基础医学
题名:
Effect of dexamethasone on peroxisome proliferator activated receptor-gamma mRNA expression in 3T3-L1 adipocytes with the human recombinant adiponectin
作者: She Qi-mei; Zhao Jing; Wang Xia-lian; Zhou Chang-man; Shi Xian-zhong
关键词: adiponectin ; transfection ; 3T3-L1 adipocytes ; peroxisome proliferator activated receptor-gamma ; dexamethasone
刊名: CHINESE MEDICAL JOURNAL
发表日期: 2007-01-20
卷: 120, 期:2, 页:155-158
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology
类目[WOS]: Medicine, General & Internal
研究领域[WOS]: General & Internal Medicine
关键词[WOS]: INDUCED INSULIN-RESISTANCE ; FATTY-ACID OXIDATION ; PPAR-GAMMA ; PROTEIN ; ADIPOSE ; MICE ; OBESITY ; PLASMA ; MUSCLE
英文摘要:

Background The fat derived protein adiponectin plays an important role in the regulation of glucose metabolism. The aim of this study was to provide the experimental basis for further investigating on adiponectin (ADPN) function. Its eukaryotic recombinant was constructed and expressed in precursor cells of 3T3-L1 adipocytes. The effects of dexamethasone on peroxisome proliferator activated receptor-gamma (PPAR-gamma) mRNA expression in 3T3-L1 cells with human recombinant adiponectin were assessed.

Methods The recombinant plasmid pMD18-T-hADPN and eukaryotic expression vector pcDNA3.1(+) were digested by two restrictive endonucleases and adiponectin and linear pcDNA3.1(+) were obtained. Then, they were ligated and translated into JM109. The recombinant pcDNA3.1(+)-hADPN so obtained was identified by digestion by restrictive endonuclease and nucleotide sequencing. The 3T3-L1 precursor cells were transfected using SuperFect Transfection Reagent (Qiagen). Furthermore, 3T3-L1 cells with human recombinant adiponectin incubated with dexamethasone (0.5 mmol/L) for 24 hours, cells were collected and total RNA was extracted. The PPAR-gamma mRNA expression was quantified by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR).

Results After eukaryotic recombinant was digested by Hind III and EcoR I, fragments of 800 bp and 5.4 kb were identified by nucleotide sequence scanning and consistent with theoretical values. Electrophoretogram of RT-PCR in 3T3-L1 precursors showed only one band in front of 250 bp, which was consistent with theoretical value 234 bp. In the 3T3-L1 cells, 3T3-L1 cells with plasmid and 3T3-L1 cells human recombinant adiponectin, treatment with dexamethasone (0.5 mmol/L) decreased PPAR-gamma mRNA expression compared to untreated controls (P < 0.01). Effect of dexamethasone on PPAR-gamma mRNA expression in 3T3-L1 cells was reversed by stably transfected human recombinant adiponectin.

Conclusion The 3T3-L1 cells stably transfected human recombinant adiponectin had increased PPAR-gamma mRNA expression. Dexamethasone suppressed PPAR-gamma mRNA expression in the 3T3-L1 cells. Effect of dexamethasone on PPAR-gamma mRNA expression in 3T3-L1 cells was reversed by stably transfected human recombinant adiponectin.

语种: 英语
WOS记录号: WOS:000243781100014
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/59615
Appears in Collections:基础医学院_解剖学与组织胚胎学系_期刊论文

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作者单位: 1.Peking Univ, Hlth Sci Ctr, Dept Anat, Beijing 100083, Peoples R China
2.Shijingshan Hosp Beijing, Dept Endocrinol, Beijing 100043, Peoples R China

Recommended Citation:
She Qi-mei,Zhao Jing,Wang Xia-lian,et al. Effect of dexamethasone on peroxisome proliferator activated receptor-gamma mRNA expression in 3T3-L1 adipocytes with the human recombinant adiponectin[J]. CHINESE MEDICAL JOURNAL,2007,120(2):155-158.
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