• Title/Summary/Keyword: Water Expression

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Functional Expression of a Dipeptide Transporter Obtained from Intestinal HT-29 Cells Using Xenopus Oocytes (장관세포인 HT-29에 존재하는 디펩티드수송체의 Xenopus oocyte에서의 발현)

  • Oh, Doo-Man;Yang, Chae-Ha
    • Journal of Pharmaceutical Investigation
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    • v.25 no.4
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    • pp.299-305
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    • 1995
  • Cloning the gene encoding a dipeptide transporter is necessary for understanding the absorption mechanism of peptides and peptide-like drugs in the gastrointestinal tract. Functional expression of a dipeptide transporter after microinjection into Xenopus laevis oocytes was performed using the mRNA purified from human intestinal HT-29 cells. Fifty nanoliters of purified mRNA (1 mg/mL) were microinjected into healthy oocytes followed by incubation for 4 days in order to express a dipeptide transporter. Functional expression was determined by a uptake assay using 10 Ci/mL $[^3H]-glycylsarcosine$, a dipeptide substate of the transporter. Seasonal variability and batch-to-batch variability were greater in summer. The usage of beveled micropipettes improves viability of oocytes at 4 days after microinjection. Expression of a dipeptide transporter in oocytes after microinjection of mRNA obtained from HT-29 cells was significantly larger than those after microinjection of water or mRNA obtained from the rabbit intestine.

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Skin Hydration Effect of Jeju Lava Sea Water (제주용암해수의 피부 보습 효과 연구)

  • Lee, Sung Hoon;Bae, Il-Hong;Min, Dae Jin;Kim, Hyoung-June;Park, Nok Hyun;Choi, Ji Hae;Shin, Jin Seob;Kim, Eun Ju;Lee, Hae Kwang
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.42 no.4
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    • pp.343-349
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    • 2016
  • Many minerals and nutrient salts are abundant in Jeju lava sea water. The objective of this study was to evaluate the skin hydration effects of Jeju lava sea water. The skin barrier serves as a protective barrier that prevents the loss of moisture. The water holding capacity and water transport of the epidermis have been proposed to be important determinants of skin hydration. Jeju lava sea water increased the mRNA expression of filaggrin and caspase-14 which is related to natural moisturizing factor (NMF) formation. Aquaporins 3 (AQP3) are proteins that facilitate the transport of water across cell membranes. Jeju lava sea water increased the mRNA expression and protein expression of AQP3. We employed a skin equivalent model to assess the efficacy of Jeju lava sea water. In a skin equivalent model, Jeju lava sea water increased the CD44 (hyaluronic acid receptor) which is related to skin hydration. From these results, we found out Jeju lava sea water maybe help to skin hydration.

The Effects of DoDamTanghapChongMungTang(Daotantanghecongmingtang) on LPS induced-Microglia and Memory Deficit Mice Model (도담탕합총명탕(導痰湯合聰明湯)이 LPS로 처리된 microglia 및 기억력 감퇴 생쥐 모델에 미치는 영향)

  • Park, Dae-Myung;Lee, Sang-Ryong;Jung, In-Chul
    • Journal of Oriental Neuropsychiatry
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    • v.22 no.2
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    • pp.107-128
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    • 2011
  • Objectives : This experiment was designed to investigate the efficacy of DDTCMT hot water extract & ultra-fine powder on Alzheimer's Disease Model. Methods : The effects of the DDTCMT hot water extract on expression of IL-$1{\beta}$, IL-6, TNF-${\alpha}$, COX-2, NOS-II, IL-10, IL-1 receptor antagonist mRNA and production of IL-$1{\beta}$, IL-6, TNF-${\alpha}$ in BV2 microglial cell line treated by lipopolysacchaide(LPS) were investigated. Expression of NO, ROS in BV2 microglial cell line treated by LPS and AChE activity in PC-12 cell treated by NGF were investigated. anti-AChE was observed through Western blot analysis. The effects of the DDTCMT hot water extract & ultra-fine powder on the behavior of the memory deficit mice induced by scopolamine were investigated. Results : 1. The DDTCMT hot water extract significantly decreased the production of mIL-6, mNOS-II, mTNF-${\alpha}$, and increased the production of mIL-10, mIL-1 receptor antagonist. 2. The DDTCMT hot water extract significantly suppressed the production of IL-$1{\beta}$, IL-6, TNF-${\alpha}$ in BV2 microglial cell line treated by LPS. 3. The DDTCMT hot water extract significantly suppressed the NO and ROS production in BV2 microglial cell line treated by LPS. 4. The DDTCMT hot water extract groups showed inhibition of AChE activity in NGF treated PC-12 cell line. 5. The DDTCMT hot water extract suppressed anti-AChE expression in NGF treated PC-12 cell line was observed by Western blot analysis. 6. The DDTCMT hot water extract & ultra-fine powder groups showed significantly inhibitory effect on the scopolamine -induced impairment of memory in the experiment of Morris water maze. Conclusions : These results suggest that the DDTCMT hot water extract & ultra-fine powder may be effective for the prevention and treatment of Alzheimer's disease.

Effects of Crataegi Fructus Hot-Water Extract on Hair Growth and Growth Factor Gene Expression of Human Dermal Papilla Cells (산사 열수추출물의 모발 성장과 모유두세포의 성장인자 유전자 발현에 대한 영향)

  • Kim, Hyeong-Key;Kim, Cheol-Hong;Yoon, Hyun-Min;Kang, Kyung-Hwa;Song, Choon-Ho
    • Korean Journal of Acupuncture
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    • v.34 no.3
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    • pp.146-155
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    • 2017
  • Objectives : This study was carried out to investigate the effects of Crataegi Fructus water extract(CFWE) on hair growth in an alopecia model of C57BL/6N mice and human dermal papilla cells(hDPCs). Methods : Six-week old mice were depilated and separated in 3 groups ; CON, MXD(2% Minoxidil), and CFWE. The treatments were applied twice a day for 18 days. The hair growth was determined photographically. The hair density, thickness and length were identified by Folliscope and the weights of body were measured. In dorsal skin tissue, the expression of hair growth-related protein was analyzed by Western blot. In hDPCs with/without $IFN-{\gamma}$, cell proliferation and the expression of hair growth-related genes were analyzed. Results : We observed that CFWE promoted hair growth compared to CON. CFWE improved the hair density, thickness and length compared to CON. CFWE increased the $Wnt/{\beta}$-catenin signaling in dorsal skin. In hDPCs, CFWE accelerated the cell proliferation and inhibited $IFN-{\gamma}$-induced hDPCs degeneration. CFWE increased the mRNA expression of ${\beta}$-catenin, Axin-2, BMP-4, FGF-7, FGF-10, and ALP compared to CON and $IFN-{\gamma}$ treated cells. Conclusions : These results suggest that CFWE has a hair regrowth activity via $Wnt/{\beta}$-catenin signaling and can be useful for the treatment of alopecia.

Studies on Immunoregulatory Effects of Bowon-tang in the Immune Cells (保元湯의 免疫調節 作用에 관한 硏究)

  • Hwang, Joo-Min;Jung, Myung;Cho, Jung-Hoon;Lim, Kyu-Sang;Yun, Young-Gab
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.28 no.4
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    • pp.92-110
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    • 2015
  • Objectives : The water extract of Bowon-tang composited with thePanax, AstragalusandGlycyrrhiza Radixhas been traditionally used for treatment of a sickly child and smallpox in oriental medicine. However, little is known about the regulatory effects of Bowon-tang on the production, expression and activity of immune mediators [nitric oxide, prostaglandin E2, inducible nitric oxide synthetase, cyclooxygenase-2], the macrophage activation factor production, the proliferation, subset expression, the killing activity, and the capping in immune cells.Methods : In this study, we investigated the effects of water extracts from Bowon-tang,Panax, AstragalusorGRin mouse immune cells or human Jurkat T cells. Each extract (25-200 ㎍/㎖)perse had no cytotoxic effect in unstimulated macrophages, but concentration-dependently regulated NO and PGE2production, iNOS expression, and COX-2 activity in mouse peritoneal macrophages with MAF stimulation. These regulatory effects were synergistically increased by their combination (Bowon-tang).Results : The extract of Bowon-tang concentration-dependently regulated T cell proliferation, CD4+and CD8+expression, and NK killing activity in mouse splenocytes and capping in Jurkat T cells.Conclusions : These results suggest that the water extract of Bowon-tang composited with thePanax, AstragalusandGRmay be useful for therapeutic drugs against a sickly constitution and immune diseases, probably by regulating the production of immune mediators.

The Effect of Guibi-tang Water Extract on Osteoclast Differentiation and Osteoblast Proliferation (귀비탕(歸脾湯)이 파골세포 분화와 조골세포 활성에 미치는 영향)

  • Choi, Kyung-Hee;Yoo, Dong-Youl
    • The Journal of Korean Obstetrics and Gynecology
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    • v.27 no.3
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    • pp.12-27
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    • 2014
  • Objectives: This study was performed to evaluate the effect of Guibi-tang water extract (GB) on osteoporosis. Methods: We examined the effect of GB on osteoclast differentiation using murine pre-osteoclastic RAW 264.7 cells treated with receptor activator of nuclear factor kappa-B ligand (RANKL). The effect of GB on osteoclast was measured by counting TRAP (+) multinucleated cells and measuring TRAP activity. The mRNA expressions of osteoclastogenesis-related genes (Cathepsin K, MMP-9, TRAP, NFATc1, MITF, TNF-${\alpha}$, IL-6, COX-2) were measured by real-time PCR. We examined the effect of GB on osteoblast proliferation, ALP activity, bone matrix protein synthesis and collagen synthesis using murine calvarial cell. Results: GB decreased the number of TRAP (+) multinucleated cells and inhibited TRAP activity in RANKL-stimulated RAW 264.7 cell. GB decreased the expression of genes related osteoclastogenesis such as Cathepsin K, MMP-9, TRAP, NFATc1, MITF, COX-2 in RANKL-stimulated RAW 264.7 cell. But GB did not decrease the expression of iNOS and increased the expression of TNF-${\alpha}$, IL-6 in RANKL-stimulated RAW 264.7 cell. These genes (iNOS, TNF-${\alpha}$, IL-6) are thought to be related with the inflammatory bone destruction. GB increased cell proliferation of rat calvarial cell and also increased ALP activity in rat calvarial cell. GB did not increase bone matrix protein synthesis but increased collagen synthesis in rat calvarial cell. Conclusions: This study suggests that GB may be effective in treating osteoporosis by inhibiting osteoclast differentiation and its related gene expression and by increasing osteoblast proliferation.

Effects of Fragaria Orientalis Water extract on Adipogenesis and Cell Differentiation in 3T3-L1 Cells (3T3-L1 지방 전구세포의 분화 및 지방 생성에 미치는 Fragaria Orientalis L. 물 추출물의 영향)

  • Moon-Yeol Choi;Mi Hyung Kim;Mi Ryeo Kim
    • The Korea Journal of Herbology
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    • v.39 no.3
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    • pp.49-56
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    • 2024
  • Objective : Obesity, which has recently been rapidly increasing in the obese population, is caused by an imbalance in energy intake and consumption. The reason why we need to manage obesity well is that the prevalence of complications such as diabetes, atherosclerosis, insulin resistance, and cardiovascular disease increases. In this study, the effect of FO (Fragaria orientalis) water extract on fat metabolism in 3T3-L1 cells was observed to develop a new anti-obesity material based on Mongolian medical books. Methods : The effect of FO extract on adipogenesis in 3T3-L1 cells was observed using DPPH scavenging, pancreatic lipase inhibitory activity, MTT analysis and Oil-red-O staining method. And the expression of proteins related to lipid metabolism was analyzed by Western blot. Results : The FO group significantly increased the DPPH radical scavenging activity at 5 mg/ml compared to the positive control BHA at 0.1 mg/ml. In oil red O staining at a safe concentration without cytotoxicity, lipid accumulation was significantly inhibited by less than 80% compared to the control group at all concentrations. Moreover, treatment of FO significantly increased the expression of proteins related to lipid metabolism, such as p-AMPK and p-ACC, in 3T3-L1 cells, and the expression of CPT-1 tended to increase in a dose-dependent manner. However, the expression of PPAR-γ was significantly decreased in a dose-dependent manner. Conclusion : These results suggest that FO water extract has a potential anti-obesity effect and are expected to be utilized in the development of materials for obesity prevention and treatment.

Inhibitory effects of Capsicum annuum L. water extracts on lipoprotein lipase activity in 3T3-L1 cells

  • Baek, Jongmi;Lee, Jaesung;Kim, Kyoungkon;Kim, Taewoo;Kim, Daejung;Kim, Cheonan;Tsutomu, Kanazawa;Ochir, Sarangowa;Lee, Kooyeon;Park, Cheol Ho;Lee, Yong-Jik;Choe, Myeon
    • Nutrition Research and Practice
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    • v.7 no.2
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    • pp.96-102
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    • 2013
  • Obesity, an intractable metabolic disease, currently has no medical treatment without side effects, so studies have been actively carried out to find natural compounds that have anti-obesity activity with minimum side effects. In this study, the anti-obesity effects of water extracts of seven Capsicum annuum L. varieties being Putgochu (Pca), Oyee gochu (Oca), Kwari putgochu (Kca), Green pepper (Gca), Yellow paprika (Yca), Red paprika (Rca) and Cheongyang gochu (Cca), were examined through the evaluation of lipoprotein lipase (LPL) mRNA expression level in 3T3-L1 cells (mouse pre-adipocytes). After capsaicin elimination by chloroform defatting, freeze-dried powder of Cca was treated to 3T3-L1 cells and anti-obesity effects were examined by determining the LPL mRNA level using the RT-PCR method. Of the primary fractions, only proven fractions underwent secondary and tertiary refractionating to determine anti-obesity effects. From seven different Capsicum annuum L., there was a significant decrease of the LPL mRNA expression level of 50.9% in Cca treatment compared to the control group. A significant decrease of the LPL mRNA expression level was shown in primary fractions (Fr) 5 (36.2% decrease) and 6 (30.5% decrease) of the Cca water extracts. Due to the impurities checked by UPLC chromatography, Fr 5 and 6 were refractionated to determine the LPL mRNA expression level. Treatment of Fr 6-6 (35.8% decrease) and Fr 5-6 (35.3% decrease) showed a significant decrease in the LPL mRNA expression level. When analyzed using UPLC, major compounds of Fr 6-6 and Fr 5-6 were very similar. Subsequently, we refractionated Fr 6-6 and Fr 5-6 to isolate the major peak for structure elucidation. Treatment of Fr 5-6-1 (26.6% decrease) and Fr 6-6-1 (29.7% decrease) showed a significant decrease in the LPL mRNA expression level. Consequently, the fractions may have a possibility to ameliorate obesity through the decrease of the LPL mRNA expression level.

Effect of Thyroid Hormone on the Gene Expression of Myostatin in Rat Skeletal Muscle

  • Ma, Yi;Chen, Xiaoqiang;Li, Qing;An, Xiaorong;Chen, Yongfu
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.2
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    • pp.275-281
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    • 2009
  • Modification of thyroid hormone levels has a profound effect on skeletal muscle differentiation, predominantly through direct regulation involving thyroid hormone receptors. Nevertheless, little is known about the regulation of myostatin gene expression in skeletal muscle due to altered concentrations of thyroid hormone. Thus, the goal of our study was to find out whether altered thyroid states could change the gene expression of myostatin, the most powerful inhibitor of skeletal muscle development. A hyperthyroid state was induced in rats by daily injections of L-thyroxine 20 mg/100 g body weight for 14 days, while a hypothyroid state was induced in another group of rats by administering methimazole (0.04%) in drinking water for 14 days. After a period of 14 days of L-thyroxine treatment we observed a significant increase of myostatin expression both in mRNA and protein level. However, decreased expression of myostatin mRNA and protein were observed in hypothyroid rats. Furthermore, our studies demonstrated that the upregulation of myostatin gene expression might be responsible for the loss of body weight induced by altered thyroid hormone levels. We concluded that myostatin played a role in a metabolic process in muscle that was regulated by thyroid hormone.