• Title/Summary/Keyword: inhibiting factor

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Inhibitory Effect of Purple Corn 'Seakso 1' Husk and Cob Extracts on Lipid Accumulation in Oleic Acid- Induced Non-Alcoholic Fatty Liver Disease HepG2 Model (올레산 유도 비알코올성 지방간세포에서 자색옥수수 색소 1호 포엽과 속대 추출물의 지질 축적 억제 효과)

  • Lee, Ki Yeon;Kim, Tae hee;Kim, Jai Eun;Bae, Son wha;Park, A-Reum;Lee, Hyo Young;Choi, Sun jin;Park, Jong yeol;Kwon, Soon bae;Kim, Hee Yeon
    • Journal of Food Hygiene and Safety
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    • v.35 no.1
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    • pp.93-101
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    • 2020
  • Seakso 1, a maize hybrid, was developed in 2008 by Gangwon Agricultural Research and Extension Services in Korea and registered in 2011. It is single-cross hybrid, semi-flint, deep-purple variety of corn, variety of are yellow, while the husks and cobs are purple. Due to the sensitivity of Seakso 1 to excess moisture after seeding, water supply should be carefully managed, and it should be harvested at a suitable time to obtain the highest anthocyanin content. This study investigated the hepatoprotective effect of Saekso 1 corn husk and cob extracts (EHCS) in oleic acid-induced non-alcoholic fatty liver disease (NAFLD) in HepG2 cells. EHCS showed a high level of lipid accumulation inhibiting effect. EHCS also suppressed triglyceride accumulation and inhibited expression of lipid marker genes, such as sterol regulatory element binding protein-1c (SREBP-1c) and sterol regulatory element binding protein-1a (SREBP-1a). Analysis by western blot of the expression of p-AMPK, p-SREBP1, PPARα, and FAS proteins showed that the incidence of SREBP1 protein, a major factor involved in lipid metabolism in the liver, has decreased significantly after treatment with the extracts. Moreover, the protein-induced expression of FAS, a major enzyme involved in the biosynthetic pathways of fatty acids, was decreased significantly in all concentrations. These results suggest that EHCS is a potent agent for the treatment of NAFLD.

Comparison of Anti-inflammatory Activities among Ethanol Extracts of Sophora flavescens, Glycyrrhiza uralensis and Dictamnus dasycarpus, and their Mixtures in RAW 246.7 Murine Macrophages (RAW 246.7 대식세포 모델에서 고삼, 감초, 백선피 에탄올 추출물 및 추출복합물의 항염증 효능 비교)

  • Han, Min Ho;Lee, Moon Hee;Hong, Su Hyun;Choi, Yung Hyun;Moon, Ju Sung;Song, Myung Kyu;Kim, Min Ju;Shin, Su Jin;Hwang, Hye Jin
    • Journal of Life Science
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    • v.24 no.3
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    • pp.329-335
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    • 2014
  • Sophora flavescens, Glycyrrhiza uralensis and Dictamnus dasycarpus have been widely used in folk medicine for several inflammatory disorders in Korea and China. In this study, we compared the anti-inflammatory effects of the ethanol extracts of S. flavescens (EESF), G. uralensis (EEGU) and D. dasycarpus (EEDS), and their mixtures (medicinal herber mixtures, MHMIXs) on production of inflammatory mediators and cytokines in lipopolysaccharide (LPS)-stimulated RAW 264.7 murine macrophages. Our data indicated that treatment with EESF, EEGU and EEDD significantly inhibited the excessive production of pro-inflammatory mediators such as nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) in LPS-stimulated RAW 264.7 cells. The ethanol extracts and MHMIXs also attenuated the production of pro-inflammatory cytokines, including interleukin-$1{\beta}$ ($IL-1{\beta}$) and tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$) by suppressing their protein expression, respectively. Interestingly, MHMIX-1, which basic ingredients are EESF, EEGU and EEDS in the proportion 3:1:1, more safely and effectively inhibits the LPS-induced inflammatory status in LPS-stimulated RAW 264.7 macrophages compared to ethanol extracts of each medicinal herb and other MHMIXs without causing any cytotoxic effects. Our study provides scientific evidence to support that a berbal mixture, MHMIX-1 may be useful in the treatment of inflammatory diseases by inhibiting inflammatory regulator responses in activated macrophages.

Inhibitory Effects of Tenebrio molitor Larvae Ethanol Extract on RANKL-Induced Osteoclast Differentiation (갈색거저리 유충 에탄올 추출물이 RANKL에 의해 유도되는 파골세포 분화에 미치는 영향)

  • Seo, Minchul;Baek, Minhee;Lee, Hwa Jeong;Shin, Yong Pyo;Lee, Joon Ha;Kim, In-Woo;Kim, Mi-Ae;Hwang, Jae-Sam
    • Journal of Life Science
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    • v.30 no.11
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    • pp.983-989
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    • 2020
  • The balance between bone-resorbing osteoclasts and bone-forming osteoblasts is key to bone health. An imbalance between osteoclasts and osteoblasts leads to various bone-related disorders, such as osteoporosis, osteomalacia, and osteopetrosis. However, the bone-resorption inhibitor drugs that are currently used may cause side effects. Natural substances have recently received much attention as therapeutic drugs for the treatment of bone health. This study was designed to determine the effect of Tenebrio molitor larvae ethanol extract (TME) on receptor activator of nuclear factor-κB ligand (RANKL)-induced osteoclast differentiation. To measure the effect of TME on osteoclast differentiation, RAW264.7 cells were treated with RANKL with or without TME for 5 days. The tartrate-resistant acid phosphatase (TRAP) activity was significantly inhibited by treatment of TME without cytotoxicity up to 2 mg/ml. In addition, TME effectively suppressed expression of osteoclast differentiation-related marker genes and proteins such as TRAP, NFATc1, and c-Src. TME also significantly inhibited the p38 mitogen-activated protein kinase (MAPK) signaling pathway without affecting ERK and JNK signaling in RANKL-induced RAW264.7 cells. Consequently, we conclude that TME suppresses osteoclast differentiation by inhibiting RANKL-induced osteoclastogenic genes expression through the p38 MAPK signaling pathways. These results suggest that TME and its bioactive components are potential therapeutics for bone-related diseases such as osteoporosis.

Antioxidant and Inhibitory Effects of Korean Panax ginseng Extract on Pro-inflammatory Mediators in LPS-stimulated RAW264.7 Macrophages (산양삼(Korean Panax ginseng) 추출물의 항산화 효과 및 LPS로 염증이 활성화된 RAW 264.7 대식세포에서의 염증매개물질 억제효과)

  • Kim, Ye-Jin;Son, Dae-Yeul
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.10
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    • pp.1371-1377
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    • 2012
  • Biological activities of Korean Panax ginseng 55% ethanol extract (KPGE) were investigated. The measured total polyphenol content of KPGE was 357.45 mg/100 g. KPGE showed the highest ${\alpha},{\alpha}$-diphenyl-${\beta}$-picrylhydrazyl (DPPH) and 2,2'-azino-bis-3-ethylbenzo-thiazoline-6-sulfonic acid (ABTS) radical scavenging activities of 80% and 86% at 1,000 ${\mu}g/mL$, respectively. DPPH and ABTS radical scavenging activities significantly increased in a KPGE concentration-dependent manner. SOD-like activity of KPGE (1, 10, and 100 ${\mu}g/mL$) increased from 22% up to 33% at KPGE concentrations of 500 and 1,000 ${\mu}g/mL$. KPGE treatment significantly suppressed the generation of pro-inflammatory mediators, including nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$), and cytokines (tumor necrosis factor-alpha: TNF-${\alpha}$, interleukin-6: IL-6, interleukin-$1{\beta}$: IL-$1{\beta}$), in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. KPGE demonstrated strong anti-inflammatory activity that reduced NO and $PGE_2$ production in LPS-stimulated RAW 264.7 cells. Even low concentrations of KPGE (1 and 10 ${\mu}g/mL$) reduced $PGE_2$ and NO production in RAW 264.7 macrophages without LPS-stimulation, respectively. At concentrations of 100, 500, and 1,000 ${\mu}g/mL$, TNF-${\alpha}$, IL-$1{\beta}$ and IL-6 production were significantly suppressed. The results of our study suggest the potential of Korean Panax ginseng as an excellent antioxidant substance for inhibiting inflammatory mediators. Therefore, Korean Panax ginseng (KPGE) may be used as a therapeutic approach to various inflammatory diseases.

Ethanol Extract of Glycyrrhiza uralensis Protects Against Oxidative Stress-induced DNA Damage and Apoptosis in Retinal Pigment Epithelial Cells (망막색소상피세포에서 감초 추출물의 산화적 스트레스에 의한 DNA 손상 및 apoptosis 유발의 차단 효과)

  • Kim, So Young;Kim, Jeong-Hwan;Kim, Sung Ok;Park, Seh-Kwang;Jeong, Ji-Won;Kim, Mi-Young;Lee, Hyesook;Cheong, JaeHun;Choi, Yung Hyun
    • Journal of Life Science
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    • v.29 no.11
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    • pp.1273-1280
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    • 2019
  • Age-related macular degeneration (AMD) is one of the leading causes of blindness in the elderly population, and damage to retinal pigment epithelial (RPE) cells due to oxidative stress contributes to the development of AMD. Glycyrrhiza uralensis Fischer is one of the most widely used herbal medicines for the treatment of various diseases in Asian countries. Although recent studies indicated that treatment with G. uralensis can protect cells from oxidative stress, its mechanisms in RPE cells remain unknown. We evaluated the effect of a G. uralensis ethanol extract (GU) on $H_2O_2$-induced oxidative injury in ARPE-19 RPE cells. The GU pretreatment attenuated reactive oxygen species (ROS) generation induced by $H_2O_2$, which was associated with induced expression of nuclear factor erythroid-derived-2-like 2 (Nrf2) and heme oxygenase-1 (HO-1). GU also suppressed $H_2O_2$-induced DNA damage and mitochondrial dysfunction. The inhibitory effect of GU on $H_2O_2$-induced apoptosis was associated with the protection of caspase-3 activation. Overall, GU appeared to protect RPE cells from oxidative injury by inhibiting DNA damage and reducing apoptosis. Further studies are needed to determine the regulation of Nrf2-mediated HO-1 expression, but our results suggest the possibility of using GU to reduce the risk of AMD.

Evaluation Antioxidant and Anti-inflammatory Activity of Ethanolic Extracts of Myriophyllum spicatum L. in Lipopolysaccharide-stimulated RAW 264.7 Cells (이삭물수세미(Myriophyllum spicatum L.) 에탄올 추출물의 항산화와 항염증 효과)

  • Chul Hwan Kim;Young-Kyung Lee;Min Jin Kim;Ji Su Choi;Buyng Su Hwang;Pyo Yun Cho;Young Jun Kim;Yong Tae Jeong
    • Korean Journal of Plant Resources
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    • v.36 no.1
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    • pp.15-25
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    • 2023
  • Myriophyllum spicatum L. has been used as an ornamental in ponds and aquariums, and as a folk remedy for inflammation and pus. Nevertheless, the biological activity and underlying mechanisms of anti-inflammatory effects are unclear. This study is aimed at investigating the antioxidative and anti-inflammatory activities of ethanol extract of Myriophyllum spicatum L. (EMS) in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. Antioxidant activity of EMS was assessed by radical-scavenging effects on ferric reducing antioxidant power (FRAP) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radicals. As inflammatory response parameters produced by LPS-stimulated RAW 264.7 cells were quantified to assess the anti-inflammatory activity of EMS. Our results showed that EMS increased FRAP and DPPH radical-scavenging activity. In EMS-treated RAW 264.7 cells, the production of NO, PGE2, TNF-α and IL-1β was significantly inhibited at the non-cytotoxic concentration. In addition, EMS significantly attenuated LPS-stimulated the toll-like receptor (TLR) 4/myeloid differentiation protein (MyD) 88 signaling pathway, and inhibited nuclear translocation of nuclear factor-kappa B(NF-κB). Positive correlations were noted between anti-inflammatory activity and antioxidant activity. In conclusion, it was indicated that EMS suppresses the transcription of inflammatory factors by inhibiting the TLR4/MyD88/NF-κB signaling pathway, thereby suppressing LPS-stimulated inflammation in RAW 264.7 cells. This study highlights the potential role of EMS against inflammation and associated diseases.

Effects of Evaporative Water-loss from Cultural Pots on Growth of Pot-grown Ornamental Plants (화분(花盆)의 수분증발(水分蒸發)이 분식화훼류(盆植花卉類)의 생육(生育)에 미치는 영향(影響))

  • Suh, Youn-gkyo
    • Korean Journal of Agricultural Science
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    • v.4 no.2
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    • pp.317-343
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    • 1977
  • This study was carried out to obtain the informations about evaporation from pot, soil temperature and soil atmosphere composition in pot, and the effect on the growth of nine ornamental species using seven different containers. The investigated containers were clay pot(CP), clay pot painted in green(CP-P), varnished clay pot(CP-V), polyethylene film inserting in clay pot(CP-PI), clay pot mulched with black polyethylene film(CP-PM), porcelain pot(POP), and plastic pot(PLP). Nine ornamental species were balsam(Impatiens balsamina), chrysanthemum(Chrysanthemum morifolium), cosmos(Cosmos bipinatus), English ivy(Hedera helix), geranium(Pelargonium zonale), kochia(Kochia scoparia var. trichophila), marigold(Tagetes patula), ornamental kale(Brassica oleraceae var. acephala), and salvia (Salvia splendens). The results obtained are summarized as follows: 1. Dry weight of all tested species grown in PLP, POP, CP-P, CP-V and CP-PI was heavier than that of CP. 2. Plant height in nine tested species grown in PLP, POP, CP-P, CP-V, and CP-PI was taller than that of CP. 3. Geranium grown in PLP, POP, CP-P, and CP-V gave more number of leaf than that of CP. 4. The number of flower in balsam grown in PLP, POP, CP-P, CP-V and CP-PI was more than that of CP. The result from marigold was very similar to this tendency. Spike length and floret number in salvia gave the same tendency, but its spike number was not different among containers used. 5. The average diurnal evaporation from PLP and POP was about 43% of that of CP. About two third of total evaporation from CP was through pot wall. 6. The evaporation rate from the slowest to the highest was PLP, POP, CP-P, CP-V, CP-PI, CP-PM and CP. Containers inhibiting evaporation through pot wall hold more soil moisture than CP from one day after water supply. 7. The more evaporative water-loss from containers gave the lower soil temperature. The variation of soil temperature among containers was higher during the day than the night. 8. The $O_2$ concentration of soil atmosphere in CP was higher than that of nonporous containers, and the difference between them was 0.40-1.12%. The range of the $O_2$ concentration 17.95~19.62%. The $CO_2$ concentration of soil atmosphere in CP was lower than that of nonporous containers, and its range was 0.59-1.76%. This deviation in soil atmosphere composition did not influenced on the growth of plants. 9. There was a possitive correlation between the amount of soil water and the plant growth. 10. Plant grown on CP gave more total nitrogen content in top growth than that on PLP. C/N ratio was somewhat low in plant on CP. From the above results, $O_2$ and $CO_2$ concentration in soil atmosphere did not gave enough deviation to the extent which affect the plant growth. The effect of soil moisture on the plant growth using different containers was the far-most significant factor from this investigation. Therefore, it was obious that the utilization of the nonporous container might save the cost for water supply and reduce the production cost of the pot-grown ornamental plant in Korea eventually.

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Distortion of the Dose Profile in a Three-dimensional Moving Phantom to Simulate Tumor Motion during Image-guided Radiosurgery (방사선수술에서 종양 움직임을 재현시킨 움직이는 팬텀을 이용하여 선량 분포의 왜곡에 대한 연구)

  • Kim, Mi-Sook;Ha, Seong-Hwan;Lee, Dong-Han;Ji, Young-Hoon;Yoo, Seong-Yul;Cho, Chul-Koo;Yang, Kwang-Mo;Yoo, Hyung-Jun;Seo, Young-Seok;Park, Chan-Il;Kim, Il-Han;Ye, Seong-Jun;Park, Jae-Hong;Kim, Kum-Bae
    • Radiation Oncology Journal
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    • v.25 no.4
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    • pp.268-277
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    • 2007
  • Purpose: Respiratory motion is a considerable inhibiting factor for precise treatment with stereotactic radiosurgery using the CyberKnife (CK). In this study, we developed a moving phantom to simulate three-dimensional breathing movement and investigated the distortion of dose profiles between the use of a moving phantom and a static phantom. Materials and Methods: The phantom consisted of four pieces of polyethylene; two sheets of Gafchromic film were inserted for dosimetry. Treatment was planned to deliver 30 Gy to virtual tumors of 20, 30, 40, and 50 mm diameters using 104 beams and a single center mode. A specially designed robot produced three-dimensional motion in the right-left, anterior-posterior, and craniocaudal directions of 5, 10 and 20 mm, respectively. Using the optical density of the films as a function of dose, the dose profiles of both static and moving phantoms were measured. Results: The prescribed isodose to cover the virtual tumors on the static phantom were 80% for 20 mm, 84% for 30 mm, 83% for 40 mm and 80% for 50 mm tumors. However, to compensate for the respiratory motion, the minimum isodose levels to cover the moving target were 70% for the $30{\sim}50$ mm diameter tumors and 60% for a 20 mm tumor. For the 20 mm tumor, the gaps between the isodose curves for the static and moving phantoms were 3.2, 3.3, 3.5 and 1.1 mm for the cranial, caudal, right, and left direction, respectively. In the case of the 30 mm tumor, the gaps were 3.9, 4.2, 2.8, 0 mm, respectively. In the case of the 40 mm tumor, the gaps were 4.0, 4.8, 1.1, and 0 mm, respectively. In the case of the 50 mm diameter tumor, the gaps were 3.9, 3.9, 0 and 0 mm, respectively. Conclusion: For a tumor of a 20 mm diameter, the 80% isodose curve can be planned to cover the tumor; a 60% isodose curve will have to be chosen due to the tumor motion. The gap between these 80% and 60% curves is 5 mm. In tumors with diameters of 30, 40 and 50 mm, the whole tumor will be covered if an isodose curve of about 70% is selected, equivalent of placing a respiratory margin of below 5 mm. It was confirmed that during CK treatment for a moving tumor, the range of distortion produced by motion was less than the range of motion itself.

Clinical Analysis According to $p21^{Waf1/Cip1}\;and\;p27^{kip1}$ Expression in Gastric Cancer (위암에서의 $p21^{Waf1/Cip1}\;and\;p27^{kip1}$ 단백 발현)

  • Kim, Sin-Sun;Park, Yong-Geun;Jun, Kyong-Hwa;Jung, Hun;Song, Gyo-Young;Kim, Jin-Joo;Chin, Hyung-Min;Kim, Wook;Park, Cho-Hyun;Park, Seung-Man;Lim, Keun-Woo;Kim, Seung-Nam;Jeon, Hae-Myung
    • Journal of Gastric Cancer
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    • v.6 no.1
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    • pp.36-42
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    • 2006
  • Purpose: The $p21^{Waf1/Cip1}$ protein Inhibits the cell cycle by Inhibiting the phosphorylation at the $G1{\rightarrow}S$ check point, and the $p27^{kip1}$ protein similarly performs the suppressor function by controlling the p27-mediated G1 arrest. In this study, we analysed the clinical status and survival rates in correlations with p21 and p27 expression patterns in gastric cancer. Materials and Methods: Between 1993 and 1997, 192 patients who underwent surgeries in Catholic Medical Center were analysed retrospectively in this study. Immunohistochemical staining was performed and if the nuclei of the tumor cells were stained, we assumed those as positive results. Statistical analysis was based on clinicopathological findings and differences in survival rates. Results: The expression rate of p27 was 28.1% and 15.6% in p21 each. The ratio of T1-2(80.0%) was significantly high in p21 (+), but the ratio of T3-4 (50.6%) was slightly high in p21 (-). There was no statistical significance regarding other factors. The results in p27 was not much different from expression rate of p21 in T-stage. In addition, p27 expression in diffuse type (91.3%) was higher than in intestinal type (62.7%) by Lauren's classification (P<0.05). Also, there was no statistical significance in other factors. In the correlation of p21 and p27, p27 was positive when p21 was positive (53.5%). Conversely, p27 was negative when p21 was negative (76.5%, p<0.05). In the p21 and p27 combination test, there was higher rate of T1-2 (87.5%) in p21 (+)/p27 (+), and higher rate of T3-4 (58.1%) in p21 (-)/p27 (-) (P<0.05). Results showed higher rate of intestinal type (100%) in p21 (+)/p27 (+), and diffuse type (87.0%) was dominant in p21 (-)/p27 (-) (P<0.05) by Lauren's classification. Moreover, there was no statistical significance in the 5-year survival rate in the expression of p21 and p27, and the 5-year survival rate was highest in the case of p21 (+)/p27 (+) without statistical significance. Conclusion: In our study, $p21^{Waf1/Cip1}\;and\;p27^{kip1}$ expressed similar patterns. The expression of $p21^{Waf1/Cip1}\;and\;p27^{kip1}$ affected the degree of invasiveness of the tumor, and. Combined examination result revealed the correlation of $p21^{Waf1/Cip1}\;and\;p27^{kip1}$ with Lauren's classification and depth of invasion of the tumor. However, we assumed that little difference between the survival rates depending on expression of $p21^{Waf1/Cip1}\;and\;p27^{kip1}$ has limited their value as predictable prognostic indicators.

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Lipopolysaccharide-induced Synthesis of IL-1beta, IL-6, TNF-alpha and TGF-beta by Peripheral Blood Mononuclear Cells (내독소에 의한 말초혈액 단핵구의 IL-1beta, IL-6, TNF-alpha와 TGF-beta 생성에 관한 연구)

  • Jung, Sung-Hwan;Park, Choon-Sik;Kim, Mi-Ho;Kim, Eun-Young;Chang, Hun-Soo;Ki, Shin-Young;Uh, Soo-Taek;Moon, Seung-Hyuk;Kim, Yang-Hoon;Lee, Hi-Bal
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.4
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    • pp.846-860
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    • 1998
  • Background: Endotoxin (LPS : lipopolysaccharide), a potent activator of immune system, can induce acute and chronic inflammation through the production of cytokines by a variety of cells, such as monocytes, endothelial cells, lymphocytes, eosinophils, neutrophils and fibroblasts. LPS stimulate the mononucelar cells by two different pathway, the CD14 dependent and independent way, of which the former has been well documented, but not the latter. LPS binds to the LPS-binding protein (LBP), in serum, to make the LPS-LBP complex which interacts with CD14 molecules on the mononuclear cell surface in peripheral blood or is transported to the tissues. In case of high concentration of LPS, LPS can stimulate directly the macrophages without LBP. We investigated to detect the generation of proinflammatory cytokines such as interleukin 1 (IL-1), IL-6 and TNF-$\alpha$ and fibrogenic cytokine, TGF-$\beta$, by peripheral blood mononuclear cells (PBMC) after LPS stimulation under serum-free conditions, which lacks LBPs. Methods : PBMC were obtained by centrifugation on Ficoll Hypaque solution of peripheral venous bloods from healthy normal subjects, then stimulated in the presence of LPS (0.1 ${\mu}g/mL$ to 100 ${\mu}g/mL$ ). The activities of IL-1, IL-6, TNF, and TGF-$\beta$ were measured by bioassaies using cytokines - dependent proliferating or inhibiting cell lines. The cellular sources producing the cytokines was investigated by immunohistochemical stains and in situ hybridization. Results : PBMC started to produce IL-6, TNF-$\alpha$ and TGF-$\beta$ in 1 hr, 4 hrs and 8hrs, respectively, after LPS stimulation. The production of IL-6, TNF-$\alpha$ and TGF-$\beta$ continuously increased 96 hrs after stimulation of LPS. The amount of production was 19.8 ng/ml of IL-6 by $10^5$ PBMC, 4.1 ng/mL of TNF by $10^6$ PBMC and 34.4 pg/mL of TGF-$\beta$ by $2{\times}10^6$ PBMC. The immunoreactivity to IL-6, TNF-$\alpha$ and TGF-$\beta$ were detected on monocytes in LPS-stimulated PBMC. Some of lymphocytes showed positive immunoreactivity to TGF-$\beta$. Double immunohistochemical stain showed that IL-1$\beta$, IL-6, TNF-$\alpha$ expression was not associated with CD14 postivity on monocytes. IL-1$\beta$, IL-6, TNF-$\alpha$ and TGF-$\beta$mRNA expression were same as observed in immunoreactivity for each cytokines. Conclusion: When monocytes are stimulated with LPS under serum-free conditions, IL-6 and TNF-$\alpha$ are secreted in early stage of inflammation. In contrast, the secretion of TGF-$\beta$ arise in the late stages and that is maintained after 96 hrs. The main cells releasing IL-1$\beta$, IL-6, TNF-$\alpha$ and TGF-$\beta$ are monocytes, but also lymphocytes can secret TGF-$\beta$.

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