• Title/Summary/Keyword: 3T3-L1 cell proliferation

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Fermented Colostrum Whey Upregulates Aquaporin-3 Expression in, and Proliferation of, Keratinocytes via p38/c-Jun N-Terminal Kinase Activation

  • Seo, Sang-Ah;Park, Hyun-Jung;Han, Min-Gi;Lee, Ran;Kim, Ji-Soo;Park, Ji-Hoo;Lee, Won-Young;Song, Hyuk
    • Food Science of Animal Resources
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    • v.41 no.5
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    • pp.749-762
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    • 2021
  • Colostrum, which contains various immune and growth factors, aids wound healing by promoting keratinocyte proliferation. Aquaporins (AQPs) are small, hydrophobic membrane proteins that regulate cellular water retention. However, few studies have examined the effect of processed colostrum whey on AQP-3 expression in human skin cells. Here, we investigated the effect of milk, colostrum, fermented milk, and fermented colostrum whey on AQP-3 expression in keratinocyte HaCaT cells. Concentrations of 100-400 ㎍/mL of fermented colostrum whey were found to induce HaCaT cell proliferation. AQP-3 was found to be expressed exclusively in HaCaT cells. AQP-3 expression was significantly increased in 100 ㎍/mL fermented colostrum whey-treated cells compared with that in controls. Moreover, fermented colostrum increased p38 mitogen-activated protein kinase (MAPK) and c-Jun N-terminal kinase (JNK) phosphorylation, but not ERK1/2 phosphorylation. Thus, our results suggest that fermented colostrum whey increased AQP-3 expression in, and the proliferation of, keratinocytes via JNK and p38 MAPK activation.

Effect of Keratinocyte Derived Exosome on Proliferation and Migration on Human Skin Keratinocyte (각질형성세포 유래 엑소좀이 피부각질형성세포의 증식과 이주에 미치는 영향)

  • Kim, Do Yoon;Yu, Ho Jin;Hwang, Dae Il;Jang, Sang Hee;Lee, Hwan Myung
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.42 no.4
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    • pp.359-366
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    • 2016
  • Exosome, a small vesicle secreted from cells, has diverse functions depending on cell origins and tissue types and plays a important role in cell viability and intercellular communication. Recently, many researchers have demonstrated the use of exosomes for the treatment of cancers and immune diseases, and the development of diagnostic biomarker. However, the secretion mechanism of exosome from skin cell and its physiological functions in skin remain unclear. Thus, this study aimed to explore whether keratinocyte-derived exosome affects proliferation and migration in HaCaTs. Exosomes were isolated from HaCaTs by ExoQuick-TC and then boiled or unbolied. Boiled and unboiled exosome induced proliferation in HaCaTs in a dose-dependant manner ($0.1{\sim}20{\mu}g/mL$), respectively. Boiled and unboiled exosome at concentration of $20{\mu}g/mL$ increased proliferation level in HaCaTs by $186.96{\pm}3.87%$ and $193.48{\pm}10.48%$ compared with control group. Unboiled exosome stimulated migration in HaCaTs in a dose-dependent manner ($0.1{\sim}20{\mu}g/mL$), which reached a maxium at concentration of $20{\mu}g/mL$ ($179.39{\pm}4.89%$ of control), but boiled exosome did not affect HaCaT migration. In addition, unboiled exosome ($0.1{\sim}20{\mu}g/mL$) dose-dependently stimulated sprout outgrowth in HaCats. These results demonstrate that in exosome from HaCaTs, heat-stable components such as lipid may induce HaCaT proliferation and heat-unstable components such as protein may stimulate migration and sprout outgrowth in HaCaTs, thereby leading to reepithelialization and skin-wound healing activities. It is concluded that exosomes from HaCaTs may be used as cosmetic materials.

Histone H3K4 Methyltransferase SET1A Stimulates the Adipogenesis of 3T3-L1 Preadipocytes (히스톤 H3K4 메칠화효소 SET1A에 의한 지방세포 분화 촉진)

  • Kim, Seon Hoo;Jung, Myeong Ho
    • Journal of Life Science
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    • v.27 no.10
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    • pp.1104-1110
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    • 2017
  • SET1A is a histone H3K4 methyltransferase that catalyzes di- and trimethylation of histone H3 at lysine 4 (H3K4). Mono-, di-, and trimethylations on H3K4 (H3K4me1, H3K4me2, and H3K4me3, respectively) are generally correlated with gene activation. Although H3K4 methylation is associated with the stimulation of adipogenesis of 3T3-L1 preadipocytes, it remains unknown whether SET1A plays a role in the regulation of adipogenesis of 3T3-L1 preadipocytes. Here, we investigated whether SET1A regulates 3T3-L1 preadipocytes' adipogenesis and characterized the mechanism involved in this regulation. SET1A expression increased during 3T3-L1 preadipocytes' adipogenesis. Consistent with the increased SET1A expression, the global H3K4me3 level had also increased on day 2 after the induction of adipogenesis in 3T3-L1 adipocytes. SET1A knockdown using siRNA in 3T3-L1 preadipocytes inhibited 3T3-L1 preadipocytes' adipogenesis, as assessed by Oil Red O staining and the expression of adipogenic genes, indicating that SET1A stimulates the adipogenesis of 3T3-L1 preadipocytes. SET1A knockdown inhibited the cell proliferation of 3T3-L1 cells during mitotic clonal expansion (MCE) via down-regulation of the cell cycle gene cyclin E1, as well as the DNA synthesis gene, dihydrofolate reductase. Furthermore, SET1A knockdown repressed peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$) expression during the late stage of adipogenesis. These results indicate that SET1A stimulates MCE and $PPAR{\gamma}$ expression, which leads to the promotion of 3T3-L1 preadipocytes' adipogenesis.

Isolation of Pediococcus Strain from Nuruk and Anti-Lipid Accumulation Effect of Ornithine-Containing Makgeolli on 3T3-L1 Cells (누룩으로부터 오르니틴 생성능을 갖는 Pediococcus 속 균주의 분리 및 오르니틴 함유 막걸리의 3T3-L1 세포의 중성지질 축적 억제 효과)

  • Yook, Jin-Seon;Oh, Suk-Heung;Kim, Su-Gon;Lee, Jo-Seph;Mun, Eun-Gyung;Cha, Youn-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.9
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    • pp.1264-1269
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    • 2015
  • To evaluate the functional effect of ornithine produced by isolated lactic acid bacteria, we examined the anti-lipid accumulation effect of ornithine produced by isolate lactic acid bacteria on 3T3-L1 cells. Lactic acid bacteria (Pediococcus strain) were isolated from nuruk, which is made from wheat, rice, and barley (whole grain, grits, or flour) by fermenting microorganisms (Aspergillus, Rhizopus, and yeasts). Pediococcus strain was identified by 16S rDNA sequencing analyses, and cells were collected by centrifugation and developed as an ornithine starter. makgeolli, an ornithine-containing Korean traditional alcoholic beverage, was made with isolated lactic acid bacteria and arginine. makgeolli was made with the help of ornithine starter using a makgeolli making kit. We evaluated the anti-proliferation effect of ornithine makgeolli on 3T3-L1 cells. To determine the anti-proliferation effect of ornithine makgeolli on preadipocytes, lipid droplets were quantified and stained with Oil Red O. makgeolli made with ornithine starter and arginine showed a 3-fold higher concentration of ornithine compared to makgeolli without starter and arginine. In the results of 3T3-L1 cell line experiment, lipid accumulation was significantly reduced by adding 0.05 mg/mL of ornithine makgeolli compare to the control (adipocyte without sample). In conclusion, ornithine makgeolli containing ornithine starter isolated from nuruk showed an anti-lipid accumulation effect with increased ornithine content without toxicity.

Thaumatin Isolated from Katemfe Fruit of Thaumatococcus daiellii Inhibits 3T3 L1 Adipocytes Differenciation (Thaumatococcus daiellii 열매 유래 토마틴의 3T3-L1 지방전구세포 분화 억제에 의한 항비만 효과)

  • Cha, Jae-Young;Jeong, Jae-Jun;Yang, Hyun-Ju;Park, Jun-Seok;Kim, Hyun-Woo;Kim, Su-Hyun;Jung, Hae-Jung
    • Journal of Life Science
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    • v.21 no.6
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    • pp.783-787
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    • 2011
  • The effects of thaumatin isolated from katemfe fruit of Thaumatococcus daiellii Benth on 3T3-L1 preadipocyte differentiation was investigated in vitro. 3T3-L1 adipocytes were treated with various concentrations of thaumatin ranging in 0-5 ${\mu}M$. Thaumatin reduced fat accumulation in differentiated 3T3-L1 adipocytes in a dose-dependent manner. 3T3-L1 cell proliferation was 97.0 and 88.3% at 1 and 3 ${\mu}M$ after 8 days of thaumatin treatment, respectively. Thaumatin showed a potent inhibitory effect on stained lipid droplets at a concentration of 3 ${\mu}M$. Thaumatin reduced triglyceride accumulation in differentiated 3T3-L1 cells in a dose-dependent manner, compared with positive control cells. This study provides basic information on the anti-obesity activity of thaumatin.

Effects of Tarak, Korean Traditional Fermented Milk, on Proliferation of Immune Cells and Melanin Biosynthesis (전통발효유 타락의 면역세포 증식 및 멜라닌 생성에의 효과 - 연구노트 -)

  • Kim, Soyoung;Choi, Yumi;Lee, Heera;Park, Jisoo;Han, Young-Sook;Ko, Seong-Hee;Jang, Sung-Sik;Kim, Soo-A;Shim, Jae-Hun;Yoon, Hyungeun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.11
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    • pp.1759-1762
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    • 2015
  • Tarak is a Korean traditional fermented milk product that is fermented by adding rice wine to milk. Tarak was produced with Lactobacillus paracasei ssp. paracasei M13-65-3 isolated from rice wine, and its effects on immune cell proliferation and melanin biosynthesis were investigated. Tarak extract significantly increased proliferation of T lymphocyte Jurkat clone E6-1 cells at concentrations from 10 to $100{\mu}g/mL$. Tarak inhibited activities of tyrosinase and ${\alpha}$-melanocyte-stimulating hormone-induced melanin biosynthesis in mouse skin B16-F10 cells at a concentration of $100{\mu}g/mL$. These results suggest that tarak might have functionalities for enhancing the immune system by increasing immune cell proliferation and regulating melanin biosynthesis.

Experimental studies of Glycine max Merr. (black bean), Triticum aestivum L. (wheat) and Oryza sativa L. (rice bran) extracts on the effects of hair growth activity and physical properties (검은콩, 밀, 쌀겨 추출물이 모발의 성장과 물리적 특성에 미치는 효과)

  • Park, Hye-Yoon;Kim, Su-Na;Kang, Byung-Ha;Lee, John-Hwan
    • Korean Journal of Oriental Medicine
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    • v.16 no.3
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    • pp.167-173
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    • 2010
  • Objects : Glycine max Merr. (black bean), Triticum aestivum L. (wheat) and Oryza sativa L. (rice bran) have been widely used for treatment of relaxion of smooth muscle, gastrointestinal hemorrhage and alopecia in Korean Traditional Medicine. In this research, we examined the effect of the extracts, obtained from EtOH extracts of 3 kinds of traditional plants, on hair growing activity of the DP6 and C3H10T1 cell and physical properties. Materials and Methods : On the basis of previous studies, three traditional plants were selected and we extracted them with ethanol. We evaluated their hairy dermal papillar cell proliferation activity and mouse mesenchymal stem cell in vitro model. Also, 3 herbal extracts were added to the normal shampoo formulation in ranges of 0.1% and we validated tensile properties and physical changes using aged hair. In this research, we compared the tensile strength, shine and color appearance between the hair (general formulation) and the hair after applying shampoo with natural extracts. To analyze the luster and color image, we use the SAMBA hardware and software made by Bossa Nova Technologies. Results : In the comparative test for tensile characteristic between the hair treated general formulation(control) and the hair applying special formulation including 3 kinds of extracts, tensile distance and energy of the latter are larger than control on average. The shine and color appearance were also increased after using shampoo including natural extracts(shine : 10.9%, color appearance: 24.12%). We observed the enhancement of hair growth activity in the DP6 and C3H10T1 cell. Especially black bean extracts had the most powerful effect in the dermal papillar cell proliferation. Conclusion : These experiments suggest that extracts of Glycine max Merr. (black bean), Triticum aestivum L. (wheat) and Oryza sativa L. (rice bran) stimulate the hair growth activity and can improve physical activities of aged hair. Shampoo product, which contains 3 kinds of natural extracts, would be used for the treatment for aged hair.

Effect of Greater Celandine (Chelidonium majus L.) on Rat Hepatic Stellate Cells with Liver Fibrosis (백굴채(白屈菜)가 간섬유화 과정 중 간성상세포에 미치는 영향)

  • Lee, Hong-Il;Kim, Young-Chul
    • The Journal of Internal Korean Medicine
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    • v.32 no.4
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    • pp.504-518
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    • 2011
  • Objectives : This study was performed to investigate the anti-fibrogenic effect of greater celandine on cultured rat hepatic stellate cells. Materials and Methods : Hepatic stellate cells (HSC-T6) were treated with various concentrations of greater celandine extract for 24, 48, and 72 hours. The extraction was done with distilled water. After the treatment, cell viability, proliferation, mRNA of the ${\alpha}SMA$, TIMP-1, TIMP-2, collagen I ${\alpha}$ 1, MMP-2, IL-6, TGF-${\beta}1$, PDGFr-${\beta}1$, Bcl-2, Bax, Bcl-xl, caspase-3, caspase-9 and the activities of SOD and catalase were measured by using MTT assay, BrdU assay, real-time PCR, superoxide dismutase assay and catalase assay. Results : The viability, proliferation, mRNA expression and synthesis of collagen of the hepatic stellate cells were inhibited as the concentration increased, which indicates the herb has an inhibitory effect on fibrogenesis of the liver by regulating the fibrosis associated genes in transcription. Conclusions : These results suggest that greater celandine would be beneficial in the treatment of fibrotic patients as well as for patients with chronic hepatitis.

Effect of the Ethanol Extract from Citrus Peels on Oxidative Damage in Alloxan-induced HIT-T15 Cell (진피 에탄올 추출물이 Alloxan에 의해 유도된 HIT-T15 세포의 산화적 손상에 미치는 영향)

  • Jung, Hee-Kyoung;Jeong, Yoo-Seok;Park, Chi-Deok;Park, Chang-Ho;Hong, Joo-Heon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.8
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    • pp.1102-1106
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    • 2010
  • This study was carried out to investigate the effect of ethanol extract from citrus peels (CP-Et) against the alloxan-induced oxidative damage on HIT-T15, Hamster pancreatic $\beta$-cell. Total polyphenol and flavonoid contents in CP-Et were $57.00{\pm}2.91\;mg/g$ and $8.11{\pm}2.83\;mg/g$, respectively. Cell toxicity on HIT-T15 by CP-Et (0.125~0.75 mg/mL) was not observed. CP-Et (0.125 mg/mL) increased cell proliferation rate of HIT-T15, which was treated alloxan ($IC_{50}=11.58\;mM$) (cell viability=$80.52{\pm}3.29%$ of normal cell, p<0.05). In comparison with insulin secretion of oxidative damaged HIT-T15, 1.5 fold ($116.93{\pm}2.11\;{\mu}g/mg$ protein) was increased by treatment CP-Et treatment (0.125 mg/mL) in HIT-T15 (p<0.05). These results showed that CP-Et contribute to repairing cells and improvement of insulin expression on oxidative stress pancreatic $\beta$-cell, and also suggested application of CP-Et as a functional food material for type 2 diabetes.

Characteristics of Purinergic Receptor Expressed in 3T3-L1 Preadipocytes

  • Lee, Hyung-Joo;Baik, Joon-Heum;Kim, Min-Jeong;Kim, Na-Hyun;Kong, In-Deok
    • Biomedical Science Letters
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    • v.15 no.4
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    • pp.319-326
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    • 2009
  • Extracellular ATP elicits diverse physiological effects by binding to the G-protein-coupled P2Y receptors on the plasma membrane. In addition to the short-term effects of extracellular nucleotides on cell functions, there is evidence that such purinergic signalling can have long-term effects on cell proliferation, differentiation and death. The 3T3-L1 cell line derived from mouse embryo is a well-established and commonly utilized in vitro model for adipocytes differentiation and function. However, the distributions and roles of P2Y subtypes are still unknown in the preadipocyte. In this study, we identified the distributions and roles of P2Y subtypes in preadipocyte using $Ca^{2+}$ imaging and realtime PCR. ATP increased the $[Ca^{2+}]_i$ in a concentration-dependent manner. ATP increased $Ca^{2+}$ in absence and/or presence of extracellular $Ca^{2+}$. Suramin, non-selective P2Y blocker, largely blocked the ATP-induced $Ca^{2+}$ response. U73122, a PLC inhibitor, completely inhibited $Ca^{2+}$ mobilization in 3T3-L1 cells. The mRNA expression by realtime PCR of P2Y subtypes was $P2Y_2:P2Y_5:P2Y_6=1.0:12.5:0.3$. In conclusion, we showed that $P2Y_5$ receptor is a dominant purinergic receptor in preadipocytes, and multiple P2Y receptors could involve in differentiation and migration via regulating of intracellular calcium concentration.

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