• Journal of Life Science
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• v.22 no.1
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• pp.49-54
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• 2012

The effects of genistein on cell proliferation and adipogenesis were examined in mouse 3T3-L1 preadipocyte cells. Genistein decreased viability of 3T3-L1 pre-adipocytes in a dose-dependent manner. Oil Red O staining of these cells also indicated that adipogenesis was inhibited by 50 ${\mu}M$ genistein treatment. We investigated the molecular mechanisms involved in the decrease in cell viability in genistein-treated 3T3-L1 cells by conducting an oligo DNA microarray analysis. We selected the sirtuin-1 gene, one of the upregulated genes, for further experimentation because sirtuin-1 belongs to the sirtuin family, which is associated with anti-obesity and anti-inflammation activities. We found that four phytochemicals (resveratrol, capsaicin, daidzein, and genistein) could increase sirtuin-1 expression. Genistein was the strongest inducer of sirtuin-1 among the tested phytochemicals. The inhibition of adipogenesis by genistein was recovered by surtuin-1 siRNA transfection. Overall, these results may further our understanding of the molecular mechanisms underlying the inhibition of proliferation and adipogenesis by genistein in mouse 3T3-L1 cells.

• Korean Journal of Plant Resources
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• v.23 no.2
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• pp.165-171
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• 2010

Oxidative stress induced by reactive oxygen intermediates has been implicated in a variety of human diseases including neurodegenerative disorders, cancer, cardiovascular and respiratory diseases, and mode of action of environmental toxicants. Tert-butylhydroperoxide (t-BHP) is an organic lipid hydroperoxide analogue, which is commonly used as a pro-oxidant for evaluating mechanisms involving oxidative stress in cells and tissues. In this study, the underlying mechanisms involved in the protective effects of Hwabuk 94 kiwifruit (Actinidia deliciosa cv. 'Hwabuk 94'), which is cultivated in Jeju, on the t-BHP-induced cytotoxicity in PC12 cell. The pretreatment of rat pheochromocytoma cell line PC12 with Hwabuk 94 extract ($1-100\;{\mu}g/ml$) resulted in a significant recovery from t-BHP-induced cell death and increased Bcl-2 and procaspase-3 expression, whereas the expression of Bax and cleaved PARP were decreased in a dose-dependent manner compared to the control. Furthermore, Hwabuk 94 inhibited the t-BHP-induced p38 MAP kinase and extracellular signal-regulated kinase 1/2, but not c-Jun N-terminal kinase activations. Finally, these findings suggest that Hwabuk 94 kiwifruit might attenuate t-BHP-induced PC12 cell cytotoxicity, at least in part, through the inhibition of signaling pathways mediated by the ERK1/2 and p38 MAP kinase.

• Journal of Applied Biological Chemistry
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• v.66
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• pp.369-378
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• 2023

Smilax sieboldii is one of the Smilax species. A number of Smilax plants have multiple physiologically-active components and anti-inflammatory/anti-oxidant effects. Antiobesity effects induced by Smilax sieboldii have not been reported. In this study, we investigated the effects and molecular mechanisms of anti-obesity activity of 70% ethanol Smilax sieboldii extract (SSE). The anti-obesity effect of SSE was determined using 3T3-L1 adipocytes. We confirmed that SSE was not cytotoxic to murine 3T3-L1 preadipocytes, we evaluated SSE dose-dependently decreased the accumulation of lipids via an Oil Red O assay and triglyceride assay. These anti-obesity activities of SSE were mediated by the inhibition of adipogenesis-related marker genes (peroxisome proliferator activated receptor-γ, CCAAT-enhancer-binding protein α, and SREBP1c) and lipogenesis-related marker genes (fatty acid synthase and aP2). These results suggest that SSE has the potential to exert anti-obesity and anti-hyperlipidemia effects by regulating adipogenic transcription factors and inhibiting the expression of adipogenic markers.

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.2
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• pp.278-286
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• 2004

The objective of the current study was to determine the effects of arabinoxylane and PSP on mouse splenocytes, T cells, B cells and macrophages in vitro. Arabinoxylane and PSP directly induced the proliferation of spleen cells in a dose-dependent manner and increased IFN-${\gamma}$ synthesis. Especially, PSP induced IL-2, IL-4 and IL-10 production, Both arabinoxylane and PSP increased PFC (plaque forming cell) and RFC (rosette forming cell) formation. Arabinoxylane was not induced the proliferation of T cells, but PSP directly induced the proliferation of T cells in a high dose. Arabinoxylane and PSP increased the proliferation of B cells and the phagocytic effects of macrophage. When arabinoxylane and PSP were used in macrophage cell line stimulation, there was a marked induction of NO synthesis in a dose-dependent and an increased TNF-$\alpha$ and IL-6 synthesis. Especially, PSP also induced IL-1$\beta$ production. When arabinoxylane and PSP treated in macrophage cell line, there was induction of MHC class II expression. These results suggest that the capacity of arabinoxylane andPSP seem to act as a potent immunomodulator causing augmentation of immune cell activity, and with the absence of notable side-effects, arabinoxylane and PSP could be used as a biological response modifier having possible therapeutic effects against immunological disorders.

• Clinical and Experimental Reproductive Medicine
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• v.30 no.4
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• pp.317-324
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• 2003

목 적: 본 실험의 목적은 자궁내막세포를 분리 및 배양법 확립과 함께 불멸화 시키는 것이다. 방 법: 자궁내막에서 상피세포(epithelial cells)와 기질세포(stromal cells)의 분리는 Satyawaroop 등(1979)의 방법에 기초를 두었다. 자궁내막에서 상피세포와 기질세포의 순수 분리도를 확인하고, 불멸화된 기질세포에서 SV40 large T antigen을 확인하기 위하여 면역형광 염색(immunocytochemistry)과 Western blot 기법을 이용하였다. 정상 기질세포의 경우 subconfluence (60%) 상태에서 transfection을 진행하였다. 순수 분리된 plasmid DNA와 Qiagen 사의 superfect를 이용하여 transfection을 실시하였다. 결 과: 본 연구에서 우리는 두 가지 형태의 자궁내막 세포의 분리 및 배양에 성공하였다. 상피세포는 다면체의 형태를 띠며, 선(grandular)조직의 조각으로부터 나선형으로 자란다.기질 세포는 길쭉한 형태를 띠며, 상피세포에 비해 오래 살고, 빠르게 증식하여 나란한 형태로 배열된 세포 다발(cell bundle)을 형성한다. 이렇게 분리된 세포들은 95%의 균질성을 보였으며, 면역형광염색과 western blot을 통해 확인 하였다. 한편 SV40(Simian Virus 40) large T 항원을 암호화 하고 있는 염기 서열을 포함한 플라스미드 벡터로 안정적인 트랜스펙션을 시킴으로써 불멸화 된 자궁내막의 기질 세포주를 확립하였다. 불멸화 된 세포는 그 세포가 유래한 정상의 세포와 동일한 표현형을 가지고 있었다. 결 론: 본 연구에서, 우리는 자궁내막에서 상피세포(epithelial cells)과 기질세포(stromal cells)를 분리하여 배양법을 확립하였다. 동시에 SV40 large T antigen을 이용하여 불멸화된 세포주를 확립하였다. 이렇게 확립된 세포주는 자궁의 생리작용 연구 및 자궁내막증(Endometriosis)과 자궁암(Endometrial cancer) 등과 같은 여러 자궁관련 질병 연구에 많은 도움이 될 것으로 사료된다.

• Journal of Life Science
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• v.22 no.8
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• pp.1052-1056
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• 2012

Undaria pinnatifada has been used as a natural diet food with few calories and as a source of iodine. Even though U. pinnatifida has been regarded as a diet food, the mechanisms of its inhibitory effects on adipocyte differentiation and the accumulation of fat in adipocytes are poorly understood. In this study, the effect and mechanism of U. pinnatifida ethanol extract on 3T3-L1 differentiation into adipocytes were investigated. The effects of U. pinnatifida ethanol extract on cell viability and the anti-adipogenic effect were investigated via MTT assay, Oil red O staining, RT-PCR, and western blot. The U. pinnatifida ethanol extract did not show toxicity up to a concentration of 50 ${\mu}g/ml$. The addition of U. pinnatifida ethanol extract decreased triglyceride contents by 40% when 50 ${\mu}g/ml$ of U. pinnatifida ethanol extract was added during 3T3-L1 differentiation and adipocyte triglyceride formation. The transcription and expression of peroxisome proliferator-activated receptor ${\gamma}$ ($PPAR{\gamma}$), leptin, and hormone-sensitive lipase (HSL) as adipocyte-specific proteins were determined by RT-PCR and western blot. The overexpression of $PPAR{\gamma}$ could accelerate adipocyte differentiation. Also, leptin was secreted for triglyceride accumulation in the adipocytes and the increase of adipocyte cell size. Thus, $PPAR{\gamma}$ and leptin were used as indicators of obesity. $PPAR{\gamma}$ and leptin were repressed by the increased addition of U. pinnatifida ethanol extract. This indicates that U. pinnatifida was effective as an anti-obesity agent by repressing the differentiation of 3T3-L1 into adipocytes and inhibiting triglyceride formation in adipocytes.

• Journal of Life Science
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• v.13 no.4
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• pp.541-550
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• 2003

E2 glycoprotein of hepatitis C virus (HCV) comprises a surface of viral particle together with E1 glycoprotein, and is thought to be involved in the attachment of HCV viral particle to receptor (s) on the permissible cells including hepatocytes, B cells, T cells, and monocytes. We constructed a phage library expressing cellular proteins of hepatocytes on the phage surface, which turned out to be 8.8${\times}$$10^5$ cfu of diversity and carried inserts in 95% of library. We screened both cDNA phage library and 12-mer peptide library to identify the cellular proteins binding to E2 protein. Some intracellular proteins including tensin and membrane band 4.1 which are involved in signal transduction of survival and cytoskeleton organization, were selected from cDNA phage library through several rounds of panning and screening. On the contrary, membrane proteins such as CCR7, CKR-L2, and insulin-like growth factor-1 receptor were identified through screening of peptide library. Phages expressing peptides corresponding to those membrane proteins were bound to E2 protein specifically as determined by neutralization of binding assay. Since it is well known that HCV can infect T cells as well as hepatocytes, we examined to see if E2 protein can bind to CCR7, a member of C-protein coupled receptor family expressed on T cells, using CCR7 transfected tells. Human CCR7 cDNA was cloned into pcDNA3.1(-) vector and transfected into human embryonic kidney cell, 293T, and expressed on the surface of the cell as shown by flow cytometer. Binding assay of E2 protein using CCR7 transfected cells indicated that E2 protein bound to CCR7 by dose-dependent mode, giving rise to the possibility that CCR7 might be a putative cellular receptor for HCV.

• Food Science and Preservation
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• v.22 no.5
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• pp.744-750
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• 2015

The present study was designed to investigate the effects of hot water and ethanol extracts of Nelumbo nucifera Gaertner flower on lipid accumulation and reactive oxygen species (ROS) production during adipogenesis in 3T3-L1 cells. 3T3-L1 preadipocytes were treated with both hot water and ethanol extracts for up to 8 days following standard induction of differentiation. Regarding anti-adipogenic activity, compared with the control, the hot water and ethanol extracts significantly inhibited lipid accumulation (37.4 and 66.6%, respectively) and ROS production (46.4 and 46.8%, respectively) during adipogenesis in 3T3-L1 cells. Treatment with hot water and ethanol extracts significantly inhibited mRNA expression of peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$) and CCAAT/enhancer-binding protein alpha ($C/EBP{\alpha}$), thereby reducing the mRNA expression of adipocyte-specific fatty acid binding protein (aP2). Moreover, both the extracts significantly inhibited mRNA expression of NADPH oxidase (NOX4). Overall, our research suggests that N. nucifera Gaertner flower extracts might be a valuable source of bioactive compounds that exhibit anti-adipogenic activity and could have applications in the field of medicine and food industry.

• 대한세포병리학회:학술대회논문집
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• 1993.06a
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• pp.27-27
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• 1993

• Applied Biological Chemistry
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• v.44 no.1
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• pp.1-6
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• 2001

To elucidate the apoptosis mechanism of Trichoplusia ni cell, fundamental studies for apoptosis induction and suppression were performed. Hygromycin B, a known inducer of apoptosis, started the inhibition of T. ni cell growth at $200\;{\mu}/ml$ concentration. Furthermore, at $400\;{\mu}/ml$ concentration, DNA fragmentation was detected on day 2 of incubation. Although both dexamethasone and sodium butyrate inhibited T. ni cell growth, DNA fragmentation was not detected by both treatments. Also, when apoptosis induced T. ni cells with $200\;{\mu}/ml$ hygromycin B were treated with caspase inhibitor (Ac-DEVD-CHO), the apoptotsis was suppressed by 36%. In addition, N-acetylcysteine, another apoptosis repressor, also inhibited the apoptosis of T. ni cells. In order to express the anti-apoptosis gene (bcl-2), T. ni cells were transiently transformed with bcl-2 and its expression was confirmed by western blot analysis. These results showed the potential of developing new insect cell lines with suppressed apoptosis.