• Title/Summary/Keyword: mouse 3T3-L1

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Site-Specific Recombination by the Integrase MJ1 on Mammalian Cell (동물 세포 내에서 MJ1 인티그라제에 의한 부위 특이적 재조합)

  • Kim, Hye-Young;Yoon, Bo-Hyun;Chang, Hyo-Ihl
    • Microbiology and Biotechnology Letters
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    • v.39 no.4
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    • pp.337-344
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    • 2011
  • Integrase MJ1 from the bacteriophage ${\Phi}FC1$ carries out recombination between two DNA sequences (the phage attachment site, attP and the bacterial attachment site, attB) in NIH3T3 mouse cells. In this study, the integration vector containing attP, attB and the integrase gene MJ, was constructed. The integration mediated by integrase MJ1 in Escherichia coli led to excision of LacZ. Therefore, the frequency of integration was measured by the counting of the white colony, which is detectable on X-Gal plates. The extrachromosomal integration in NIH3T3 mouse cells was monitored by the expression of the green fluorescent protein (GFP) as a reporter. To demonstrate integration mediated integrase MJ1 in NIH3T3 cells, vectors containing attP and attB were co-transfected into NIH3T3 cells. The integration was confirmed by fluorescent microscopy. The expression of GFP was induced in NIH3T3 cells expressing MJ1 without accessory factors. By contrast, the excision mediated by the MJ1 between attR and attL had no effect on the expression of GFP. These results suggest that integrase MJ1 may enable a variety of genomic modifications for research and therapeutic purposes in higher living cells.

Artemisia scoparia Inhibits Adipogenesis in 3T3-L1 Pre-adipocytes by Downregulating the MAPK Pathway (비쑥 추출물이 3T3-L1 지방세포 분화 및 MAPK 신호 전달 경로에 미치는 영향)

  • Oh, Jung Hwan;Karadeniz, Fatih;Seo, Youngwan;Kong, Chang-Suk
    • Journal of Life Science
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    • v.28 no.9
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    • pp.999-1006
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    • 2018
  • Obesity is epidemic worldwide and has reportedly been linked to the progression of several metabolic and cardiovascular diseases. The natural products are decreasing the side effects of medicines used for obesity and also have health benefits dut to their numerous bioactive compounds. In this context, Artemisia scoparia is a widespread plant that has been suggested as possessing various types of bioactivity. In this study, the crude extract from A. scoparia (ASE) was tested for its ability to suppress adipogenesis in mouse 3T3-L1 pre-adipocytes. The molecular pathway by which ASE affects differentiation of 3T3-L1 cells was also investigated. The introduction of ASE to differentiating 3T3-L1 pre-adipocytes resulted in suppressed adipogenesis, as confirmed by decreased intracellular lipid accumulation. The differentiating cells treated with 10 and $100{\mu}g/ml$ of ASE showed 21.9 and 29.0% less lipid accumulation, respectively, than untreated adipocytes. In addition, the results indicated that ASE treatment lowered the expression of the adipogenesis-related factors $PPAR{\gamma}$, $C/EBP{\alpha}$, and SREBP-1c. Furthermore, treating with ASE notably decreased levels of phosphorylated p38, ERK, and JNK in 3T3-L1 adipocytes. These results indicate that ASE exhibits significant anti-adipogenesis activity by downregulating the MAPK and $PPAR{\gamma}$ pathways during the differentiation of 3T3-L1 pre-adipocytes. Therefore, A. scoparia may be a potential source of natural products against obesity.

Inhibitory Effects of Bogigambi-tang on the Obese-mouse Induced by High Fat Diet (보기감비탕(補氣減肥湯)이 고지방사료 식이로 비만이 유발된 생쥐에게 미치는 영향)

  • Choi, Jung-Sik;Oh, Min-Seok
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.21 no.3
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    • pp.634-641
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    • 2007
  • In order to investigate the effects of Bogigambi-tang(here in after referred to BGGBT) on the obese gene and obese inhibitory, C57BL/6 mice were induced by high fat diet. C57BL/6 mice were divided into three groups(normal, high fat diet with control, high fat diet with BGGBT extract) and fed for 15 weeks. Items of this experimental study are as follows. Body weight change, final inclose of body weight, the weight change of the adipocytes in body, the level change of ALT, AST, total cholesterol, LDL-Cholesterol, triglyceride, glucose, free fatty acid and creatinine, the expression of ${\beta}$3AR and leptin gene in primary adipocytes, the production change of leptin in primary adipocytes, the expression of ${\beta}$3AR and leptin in adipocytes tissue. The following results have been obtained All experimental group have shown that the weight and the final increase of weight have decreased considerably. All experimental group have shown that the amount of the adipocyte in weight has decreased considerably. All experimental group have shown that the amount of leptin has decreased considerably. All experimental group have shown that the revelation of ${\beta}$3AR in primary adipose cell and 3T3-L1 cell has increased considerably, and that the revelation of leptin in primary adipose cell and 3T3-L1 cell has decreased considerably, All experimental group have shown that the size of adipocyte in adipocytes tissue has decreased. The high density group have shown that the adipose vacuoles in liver tissue has decreased considerably, and that the cell nucleuses has similar with normal group.

Isolation and Structure Elucidation, Molecular Docking Studies of Screlotiumol from Soil Borne Fungi Screlotium rolfsii and their Reversal of Multidrug Resistance in Mouse Lymphoma Cells

  • Ahmad, Bashir;Rizwan, Muhammad;Rauf, Abdur;Raza, Muslim;Azam, Sadiq;Bashir, Shumaila;Molnar, Joseph;Csonka, Akos;Szabo, Diana
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.4
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    • pp.2083-2087
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    • 2016
  • A new compound namely (13-(3,3-dihydroxypropyl)-1,6-dihydroxy-3,4-dihydro-1H-isochromen-8(5H)-one (1) was isolated from an ethyl acetate extract of the borne fungi Screlotium rolfsii. Its chemical structure was elucidated by spectroscopic analysis. Screlotiumol 1 were evaluated for their effects on the reversion of multidrug resistant (MDR) mediated by P-glycoprotein (P-gp) of the soil borne fungi. The multidrug resistant P-glycoprotein is a target for chemotherapeutic drugs in cancer cells. In the present study rhodamine-123 exclusion screening test on human mdr1 gene transfected mouse gene transfected L5178 and L5178Y mouse T-cell lymphoma which showed excellent MDR reversing effect in a dose dependent manner against mouse T-lymphoma cell line. Moreover, molecular docking studies of compound-1 also showed better results as compared with the standard. Therefore the preliminary results obtained from this study suggest that screlotiumol 1 could be used as a potential agent for the treatment of cancer.

Peanut sprout ethanol extract inhibits the adipocyte proliferation, differentiation, and matrix metalloproteinases activities in mouse fibroblast 3T3-L1 preadipocytes

  • Kim, Woo Kyoung;Kang, Nam E;Kim, Myung Hwan;Ha, Ae Wha
    • Nutrition Research and Practice
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    • v.7 no.3
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    • pp.160-165
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    • 2013
  • 3T3-L1 preadipocyte were differentiated to adipocytes, and then treated with 0, 10, 20, and $40{\mu}g/mL$ of peanut sprout ethanol extract (PSEE). The main component of PSEE is resveratrol which contained 5.55 mg/mL of resveratrol. The MTT assay, Oil-Red O staining, glycerol-3-phosphate dehydrogenase (GPDH) activity, and the triglyceride concentration were determined in 3T3-L1 cells. MMP-2 and MMP-9 activities as well as mRNA expressions of C/EBP ${\beta}$ and C/EBP ${\alpha}$ were also investigated. As the concentration of PSEE in adipocytes increased, the cell proliferation was decreased in a dose-dependent manner from 4 days of incubation (P < 0.05). The GDPH activity (P < 0.05) and the triglyceride concentration (P < 0.05) were decreased as the PSEE treatment concentration increased. The mRNA expression of C/EBP${\beta}$ in 3T3-L1 cells was significantly low in groups of PSEE-treated, compared with control group (P < 0.05). The MMP-9 (P < 0.05) and MMP-2 (P < 0.05) activities were decreased in a dose-dependent manner as the PSEE concentration increased from $20{\mu}g/mL$. In conclusion, it was found that PSEE has an effect on restricting proliferation and differentiation of adipocytes.

The effect of TakliSodoksan extract on anti-tumor action and immune-function (托裡消毒散이 抗腫瘍 및 免疫作用에 미치는 效果)

  • Choi, Woong;Choi, Jung-hwa
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.12 no.1
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    • pp.79-98
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    • 1999
  • The purpose of this Study was to investigate effect of TakliSodokSan(TSS) on the anti-tumor, immunocytes and nitric oxide(NO) production from mice peritoneal macrophages. This Study estimated the proliferation of L1210 cell lines, A431 cell lines, Hep-G2 cell lines, K562 cell lines, 3T3 cell lines, mouse thymocytes and mouse splenocytes and NO production from pcritoneal macrophages in vitro, and estimated the proliferation of L1210 cells, thymocytes and splenocytcs, NO production from peritoneal macrophages and body weight in L1210 cells-transplanted mice in vivo. The results were obtained as follows; 1. TSS inhibited significantly the proliferation of L1210, A431, Hep-G2, K562 cell lines in vitro. 2. TSS accelerated the proliferation of mice thymocytes and splenocytes in vitro. 3. TSS was not increased the nitric oxide production from mice peritoneal macrophages in vitro. 4. TSS inhibited significantly the proliferation of L1210 cells in Ll210 cells∼transplanted mice. 5. TSS accelerated the proliferation of mice thymocytes and splenocytes In L1210 cells-transplanted mice. 6. TSS was increased significantly the nitric oxide production from mice peritoneal macrophages in L1210 cells-transplanted mice. 7. TSS was increased the body weight as comparing with control group in Ll210 cells-transplanted mice.

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The Effects of Herbal Extract Mixture on Anti-Obesity (항비만에 대한 생약복합물의 효과)

  • Chin, Hwi-Seung;Pack, Keum-Ju;Pack, Seung-Hee;Kim, Jae-Ki
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.1
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    • pp.32-38
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    • 2009
  • In this study, we investigated the anti-obesity activity of the herbal extract mixture (HEM). The inhibitory effect of HEM on triglyceride accumulation of 3T3-L1 preadipocyte was examined by Oil-Red O staining. HEM inhibited the triglyceride accumulation of 3T3-L1 preadipocyte cell and reduced glycerol-3-phosphate dehydrogenase (GPDH) activity. We further investigated the effect of HEM in prevention of obesity in male ICR mouse for 5 weeks. Experimental groups were divided into high fat diet group (HFD), HFD supplemented with 100 mg/kg HEM group (HEM1) and HFD supplemented with 200 mg/kg HEM group (HEM2). Body weight and food efficiency ration of HEM1 and HEM2 was decreased by 52% and 50% and by 45% and 50%, respectively. The amount of adipocyte in body weight was decreased. Blood triglyceride and total cholesterol of HEM1 was significantly decreased. These results indicate that HEM may be useful in preventing obesity.

Propranolol attenuates calorie restriction- and high calorie diet-induced bone marrow adiposity

  • Baek, Kyunghwa;Park, Hyun-Jung;Hwang, Hyo Rin;Baek, Jeong-Hwa
    • BMB Reports
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    • v.47 no.10
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    • pp.587-592
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    • 2014
  • We investigated the effects of ${\beta}$-adrenergic activation on bone marrow adiposity and on adipogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). C57BL/6 mice were subjected to a control (CON), high calorie (HIGH) or low calorie (LOW) diet for 12 weeks. In each group, mice were treated with vehicle (VEH) or propranolol. The number of adipocytes per area bone marrow was increased in LOWVEH and HIGHVEH mice compared with CONVEH mice, which was attenuated by propranolol. Isoproterenol increased lipid droplet accumulation and adipogenic marker gene expression in 3T3-L1 preadipocytes and mouse BMSCs, which were blocked by propranolol. Conditioned medium obtained from MC3T3-E1 osteoblasts suppressed adipogenic differentiation of 3T3-L1 cells, which was significantly attenuated by treatment of MC3T3-E1 cells with isoproterenol. These data suggest that ${\beta}$-adrenergic activation enhances bone marrow adipogenesis via direct stimulation of BMSCs adipogenesis and indirect inhibition of osteoblast anti-adipogenic potential.

Effect of Eriobotrya folium on Local Fat via Regulation of Lipase Secretion (비파엽추출물의 지방분해효소 조절을 통한 국소 지방분해 효능)

  • Lee, Woonkyeu;Choi, You Yeon;Yang, Woong Mo
    • Journal of Korean Medicine for Obesity Research
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    • v.17 no.2
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    • pp.101-110
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    • 2017
  • Objectives: In this study, the lipolytic effects of Eriobotrya folium extract (EFE) on local fat was investigated in high fat diet (HFD)-induced obesity mouse and 3T3-L1 adipocytes. Methods: C57BL/6J mice (5 weeks) were fed HFD for 6 weeks to induce obesity. EFE (20 mg/ml, $100{\mu}l$) or saline ($100{\mu}l$) as a normal control was injected into left inguinal fat pad region, 3 times per a week for last 2 weeks. After sacrifice, body weight, and histological changes of the inguinal fat pad were evaluated. The expressions of hormone-sensitive lipase (HSL) and adipose triglyceride lipase (ATGL) in inguinal fat pad were analyzed by Western blotting. Also, lipid accumulation and lipases release were determined in 3T3-L1 adipocytes by oil red o staining. Results: EFE significantly reduced the weight of inguinal fat pad and the size of adipocytes in HFD-induced obesity mice compared to control. The treatment of EFE up-regulated the expressions of HSL and ATGL in inguinal fat pads of obesity mice, as well as 3T3-L1 adipocytes. In addition, EFE inhibited the lipid accumulation in 3T3-L1 adipocytes in a dose dependent manner. Conclusions: EFE showed lipolytic effect on local fat of HFD-induced obesity mice by up-regulation of the lipases secretion. This suggests that EFE could be considered as anti-obese substance with lipolytic property on local fat.

Korean Red Ginseng extract and ginsenoside Rg3 have anti-pruritic effects on chloroquine-induced itch by inhibition of MrgprA3/TRPA1-mediated pathway

  • Lee, Wook-Joo;Kim, Young-Sik;Shim, Won-Sik
    • Journal of Ginseng Research
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    • v.42 no.4
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    • pp.470-475
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    • 2018
  • Background: It was previously found that Korean Red Ginseng water extract (KRGE) inhibits the histamine-induced itch signaling pathway in peripheral sensory neurons. Thus, in the present study, we investigated whether KRGE inhibited another distinctive itch pathway induced by chloroquine (CQ); a representative histamine-independent pathway mediated by MrgprA3 and TRPA1. Methods: Intracellular calcium changes were measured by the calcium imaging technique in the HEK293T cells transfected with both MrgprA3 and TRPA1 ("MrgprA3/TRPA1"), and in primary culture of mouse dorsal root ganglia (DRGs). Mouse scratching behavior tests were performed to verify proposed antipruritic effects of KRGE and ginsenoside Rg3. Results: CQ-induced $Ca^{2+}$ influx was strongly inhibited by KRGE ($10{\mu}g/mL$) in MrgprA3/TRPA1, and notably ginsenoside Rg3 dose-dependently suppressed CQ-induced $Ca^{2+}$ influx in MrgprA3/TRPA1. Moreover, both KRGE ($10{\mu}g/mL$) and Rg3 ($100{\mu}M$) suppressed CQ-induced $Ca^{2+}$ influx in primary culture of mouse DRGs, indicating that the inhibitory effect of KRGE was functional in peripheral sensory neurons. In vivo tests revealed that not only KRGE (100 mg) suppressed CQ-induced scratching in mice [bouts of scratching: $274.0{\pm}51.47$ (control) vs. $104.7{\pm}17.39$ (KRGE)], but also Rg3 (1.5 mg) oral administration significantly reduced CQ-induced scratching as well [bouts of scratching: $216.8{\pm}33.73$ (control) vs.$115.7{\pm}20.94$ (Rg3)]. Conclusion: The present study verified that KRGE and Rg3 have a strong antipruritic effect against CQ-induced itch. Thus, KRGE is as a promising antipruritic agent that blocks both histamine-dependent and -independent itch at peripheral sensory neuronal levels.