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the Differentiation-and Proliferation-Inhibitory Effects of Sporamin from Sweet Potato in 3t3-l1 preadipocytes

Agricultural Sciences in China 2009年第6期8卷 671-677页

作者： XIONG Zhi-dong lI Peng-gao MU tai-hua laboratory of Food Chemistry and Nutrition Science Institute of Agro-Food Science and Technology Chinese Academy of Agricultural Sciences Beijing 100193 P.R.China Department of Nutrition and Food Hygiene School of Public Health and Family Medicine Capital Medical University Beijing 100069 P.R. China

the aim of this study was to investigate the effect of different concentrations of sporamin on the differentiation and proliferation of 3t3-lI preadipocytes, providing the theoretical basis for the development of food to treat obesity and diabetes, the isolation and purification of sporamin from sweet potato species 55-2 were performed by ammonium sulphate precipitation in combination with ion-exchange and gel filtration chromatography. With berberine as a positive control, different concentrations ofsporamin （0.000, 0.125, 0.025, 0.250, 0.500, and 1.000 mg·ml^-1 were used to treat 3t3-l1 preadipocytes. Intracellular fat accumulation and the degree of adipogenesis were quantified using Oil Red O staining and colorimetry, preadipocytes differentiation was measured by 3（4,5-dimethylthiazolyl-2-yl）-2,5-diphenyltetrazolium bromide （Mtt） spectrophotometric assay. two sporamin proteins, which were separated into sporamin A （31 kD） and sporamin B （22 kD）, could be purified by ion-exchange and gel filtration chromatography. After being treated by different concentrations of sporamin, the differentiation of 3t3-l1 preadipocytes was significantly inhibited, compared with the positive control. When the sporamin solution concentration was 0.500 mg ml-1, the accumulation of lipid droplets within the cells was significantly decreased and the optical density （OD） value of the solution from destained Oil Red O reached to 0.35, which was the lowest value （P〈 0.05）. the proliferation of 3t3-l1 preadipocytes was significantly inhibited by treating at higher sporamin concentrations. In addition, the inhibitory effect was more obvious with the prolonged treatment time （P〈 0.05）. the differentiation and proliferation of 3t3-l1 preadipocytes could be inhibited significantly by the addition of higher concentration sporamin. It was, therefore, suggested that the sporamin was potentially effective for weight loss.

关键词： sweet potato sporamin 3t3-l1 preadipocytes

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Imipramine Inhibits Adipogenic Differentiation in Both 3t3-l1 Preadioocvtes and Mouse Marrow Stromal Cells

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Journal of Genetics and Genomics 2012年第4期39卷 173-180页

作者： Haifang li Chi-chun Fong Yao Chen Guoping Cai Mengsu Yang Department of Biology and Chemistry City University of Hong Kong83 Tat Chee AvenueKowloonHong Kong SARChina Department of Biological Sciences and Biotechnology Tsinghua UniversityBeijing 100084China Shenzhen Key laboratory of Biochip Research City University of Hong KongShenzhen 518057China

Imipramine （IM） has been widely used clinically for the treatment of mental disorders. Its actions on tissues or organs other than the nervous system also need to be understood for its proper clinical use. In this study, the effects of IM on adipogenic differentiation in both 3t3-l1 preadipocytes and mouse marrow stromal cells （MSCs） were investigated. the results showed that fewer adipocytic cells were developed from 3t3-l1 preadipocytes in the presence of 0.001 to 1 μmol/l of IM as compared to control. Similar inhibitory effect was also observed in mouse MSCs. the decrease in the formation of adipocytes was accompanied with significant down-regulation at mRNA expression of the early adipogenic transcription factor, peroxisome proliferator-activated receptor γ2 （PPARγ2）. Western blot analysis further revealed that the protein expression of PPARγ2 was reduced markedly in ceils treated with IM at concentrations of 0.01, 0.1 and 1 μmol/l, suggesting that the suppression on PPAR72 was involved in IM＇s inhibition on MSCs adipogenesis. Moreover, IM at the above concentrations could stimulate the mRNA expression of β2-adrenergic receptor （AR） and β3-AR, which implicated that the effect of IM on adipogenic differentiation was partially mediated by β-ARs. Our results demonstrated for the first time that the conventional antidepressive imipramine exerts accompanied inhibitory effect on adipocyte formation, which may have possible clinical implications.

关键词： Imipramine Adipogenic differentiation 3t3-l1 preadipocytes Marrow stromal cells (MSCs)

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Verapamil inhibits 3t3-l1 preadipocyte differentiation

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Journal of Nanjing Medical University 2009年第6期23卷 403-409页

作者： Nan Gu Shi liu Xirong Guo li Fei Xiaoqin Pan Mei Guo Ronghua Chen Department of Pediatrics Nanjing Maternal & Child Health Hospital of Nanjing Medical University Nanjing 210004 China Department of Pediatrics Huaian Maternal & Child Health Hospital Huaian 223002 China Pediatric Research Institute Nanjing Medical University Nanjing 210029 China

Objective： to investigate the effect of the calcium channel blocker verapamil on adipocyte differentiation and its mechanism of action. Methods： preadipocytes from 3t3-l1 strain mouse embryos were cultured and differentiated into matured adipocytes in vitro. Verapamil was added to the culture medium in the concentration of 30 μmol/l on Day 0. Cell differentiation was determined by Oil Red O staining and marker gene mRNA expression was evaluated and compared by Rt-PCR. the fluo-3/AM probe and laser scanning confocal microscopy were used to measure intracetlular calcium concentrations. Results：（1）the differentiation rate of 3t3-l1 preadipocytes exposed to verapamil was lower than that of untreated cells. （2）Verapamil promoted the retention of pref-1 gene expression. lipoprotein lipase expression in the verapamil group was significantly lower than that in the control group on Day 4, Day 6 and Day 8 （P 〈 0.05） and resistin expression was significantly lower than that in the control group on Day 6, Day 8 and Day 10 （P 〈 0.05）. Fatty acid synthase expression in the verapamil group was significantly lower than that in the control group from Day 2 （P 〈 0.05）. （3） Intracellular concentrations of calcium [Ca^2＋]i in the verapamil group were significantly decreased compared with those in the control group on Day 2, Day 4 and Day 6 （P 〈 0.05）, while there was no obvious difference between the two groups on Day 0 （P 〉 0.05）. Conclusion： In 3t3-l1 preadipocytes verapamil significantly reduced adipocyte differentiation, down-regulated the mRNA expression of three marker genes for adipocytes differentiation, and prolonged the mRNA expression of an inhibitor of differentiation. the inhibitory effect of verapamil on differentiation may involve its role as a blocker of calcium influx in adipocytes.

关键词： verapamil 3t3-l1 preadipocytes cell differentiation calcium

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