• Title, Summary, Keyword: cytokine

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Hepatitis B Virus-Induced TNF-a Expression in Hepa-lc1c7 Mouse Hepatoma Cell Line (마우스 Hepa-1c1c7 세포주에서 B형 간염 바이러스에 의한 tumor necrosis factor-a의 발현 유도)

  • Yea Sung Su;Jang Won Hee;Yang Young-Il;Lee Youn Jae;Kim Mi Seong;Seog Dae-Hyun;Park Yeong-Hong;Paik Kye-Hyung
    • Journal of Life Science
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    • v.15 no.1
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    • pp.38-44
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    • 2005
  • Infection with hepatitis B virus (HBV) is a major health problem worldwide. Although a tremendous amount has been known about HBV, there have been obstacles in the study of HBV due to the narrow host range of HBV limited to humans and primates. In the present study, we investigated the susceptibility to HBV infection of mouse hepatoma cell line, Hepa-1c1c7. In addition, based on that human hepatocytes infected by HBV increase the expression of the pro-inflammatory cytokine TNF-a, the inducibility of TNF-a expression by HBV in the cells was determined. HBV surface antigen (HBsAg) secretion was measured by the microparticle enzyme immunoassay and steady state mRNA expression was analyzed by quantitative competitive RT-PCR. Transient transfection of Hepa-1c1c7 cells with HBV expression vector resulted in a dose-dependent induction of TNF-a expression. Infection of Hepa-1c1c7 cells with the serum of HBV carrier also increased TNF-a mRNA expression. Both in the transfected and infected cells, HBV mRNA was expressed and significant HBsAg secretion was detected. There was no significant variation in $\beta-actin$ mRNA expression by HBV. These results demonstrate that HBV is infectious to Hepa-lc1c7 in vitro and the viral infection induces TNF-a expression, which suggests that Hepa-lc1c7, a mouse hepatoma cell line, may be a possible model system for analysis of various molecular aspects of HBV infection.

The Comparison of the Effect of Cigarette and Stop Smoking-aiding Cigarette on Release of IL-6 from Bronchial Epithelial Cell (일반담배(Cigarette)와 금연 보조 담배(금연초, 허브담배, 쑥 담배)의 기관지 상피세포에서 IL-6유리 효과비교)

  • Kim, Myoung Chan;Jung, Jeil;Jung, Jong Hoon;Kim, Hak Ryul;Yang, Sei Hoon;Jeong, Eun Taik;Kim, Hui Jung
    • Tuberculosis and Respiratory Diseases
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    • v.59 no.5
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    • pp.530-535
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    • 2005
  • Background and Aims : Cigarette smoking induces an inflammatory response in the airways, which may play a key role in the pathogenesis of chronic obstructive pulmonary disease. Interleukin-6 (IL-6) is one of the cytokines that plays an important role in inducing bronchial inflammation. The aim of this study was to determine if the level of the pro-inflammatory cytokine, Interleukin-6, is increased when the bronchial epithelial cells are exposed to a cigarette smoke extract (CSE) and an extract from stop smoking-aiding cigarettes, and examined the safety of these commercially available stop smoking-aiding cigarettes. Method : Bronchial epithelial cells were exposed to CSE from cigarette and stop smoking-aiding cigarettes for 24 hours. ELISA was used to measure the IL-6 levels in the supernatant from each condition. The IL-6 mRNA levels were measured by Taqman Real time RT-PCR. N-acetyl-L-cysteine(NAC) was added to each condition to determine if NAC can inhibit the release of IL-6 from the bronchial epithelial cells when they are exposed to CSE from cigarette and stop smoking-aiding cigarettes. Result : When bronchial epithelial cells were exposed to a CSE from cigarettes and stop smoking-aiding cigarettes, each type of CSE stimulated IL-6 production from the bronchial epithelial cells. The IL-6 mRNA level in the Bronchial epithelial cells was also elevated and NAC was found to inhibit the release of IL-6 from bronchial epithelial cells when they were exposed to the CSE from cigarettes and stop smoking-aiding cigarettes. Conclusion : Commercially available stop smoking-aiding cigarette can induce bronchial inflammation and can be harmful to smokers. Therefore, the safety of these cigarettes for smoking cessation should be evaluated.

The Change of Transforming Growth Factor ${\beta}1(TGF-{\beta}1)$ Expression by Melatonin in Irradiated Lung (방사선조사된 폐에서 Melatonin에 의한 TGF-${\beta}1$ 발현의 변화)

  • Jang, Seong-Soon;Choi, Ihl-Bohng
    • Radiation Oncology Journal
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    • v.23 no.3
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    • pp.161-168
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    • 2005
  • Purpose: The changed expressions of $TGF-{\beta}1$, as a key cytokine in the fibrotic process, due to melatonin with potent antioxidative effects, were investigated in the irradiated lung using fibrosis-sensitive C57BL/6 mice. Materials and Methods: Female C57BL/6 mice were divided into control irradiation-only, and melatonin (300 mg/kg i.p. 1 hr before irradiation) pretreatment groups. The thoraces of the mice were irradiated with a single dose of 12 Gy. The mRNA expressions of $TGF-{\beta}1$ in the lung tissue 2 and 4 weeks after irradiation were quantified using semiquantitive RT-PCR, and the cellular origin and expression levels of $TGF-{\beta}1$ protein were identified using immunohistochemical staining. Results: The relative mRNA expression levels in the irradiation-only and melatonin pretreatment groups 2 and 4 weeks after irradiation were 1.92- and 1.80-fold (p=0.064) and 2.38- and 1.94-fold (p=0.004) Increased, respectively compared to those in the control group. increased expressions of $TGF-{\beta}1$ protein were prominently detected in regions of histopathologicai radiation injury, with alveolar macrophages and septal epithelial cells serving as important sources of $TGF-{\beta}1$ expression. At 2 and 4 weeks after irradiation, the expression levels of protein were $15.8\%\;vs.\;16.9\%$ (p=0.565) and $36.1\%\;vs.\;25.7\%$ (p=0.009), respectively. Conclusion: The mRNA and protein expressions of $TGF-{\beta}1$ in the lung tissue following thoracic irradiation with 12 Gy were significantly decreased by melatonin pretreatment at 4 weeks. These results indicate that melatonin may have a possible application as an antifibrotic agent in radiation-induced lung injury.

Chest X-ray Findings and Serum Tumor Necrosis Factor-αLevels in Patients with Kawasaki Disease (가와사끼병 환아에서 흉부 X-선 검사의 변화와 혈중 Tumor Necrosis Factor-α에 대한 연구)

  • Kim, Ji Young;Kwon, Jung Hyun;Kim, Kyung Hyo;Yu, Jung Hyun;Hong, Young Mi
    • Korean Journal of Pediatrics
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    • v.48 no.5
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    • pp.534-538
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    • 2005
  • Purpose : Kawasaki disease(KD) is a multisystemic inflammatory vasculitis of unknown etiology. Many complications other than cardiovascular involvement have been recognized in KD. However, there have been few reports published concerning involvement of the lungs in this disease. The purpose of this study was to examine the relationship between serum TNF-${\alpha}$, the degree of coronary artery dilatation and chest X-ray(CXR) findings. In addition, we have investigated serum anti-Mycoplasma antibody(AMA) titers in patients with KD who have abnormal CXR findings. Methods : Eighty four patients with KD were included in this study(group I; 41 patients with normal CXR fndings, group II; 43 patients with abnormal CXR findings). Serum levels of TNF-${\alpha}$ and AMA titer were measured. Results : We reviewed the CXR findings and clinical courses of 84 patients with Kawasaki disease and found abnormal CXR findings in 43 patients(51.2 percent). Peribronchial cuffing was the most frequent abnormality(22.4 percent). In the group with abnormal CXR findings(group II), a statistical difference was not noted in age, sex, duration of fever, hemoglobin, WBC, platelet, ESR, and CRP levels and incidence of coronary arterial lesions as compared with the group having normal CXR findings(group I). No difference was noted in serum TNF-${\alpha}$ level between group I and group II. 2 patients(12.5 percent) of 16 KD patients with abnormal CXR findings have positive AMA titer(above 1 : 320). Conclusion : Most of the abnormal CXR findings in KD patients were peribronchial cuffing. The abnormal CXR findings in KD patients did not mean severe inflammations. It is difficult to consider that CXR abnormalities are related to coronary arterial lesions. In addition, further study on the relationship between Mycoplasma infection and Kawasaki disease is needed.

Enhancement of Immune Activities of Ephedrae Herba and Rubi Fructus at Low Temperature Extraction (저온 추출 공정에 의한 마황과 복분자의 면역 활성 증진 효과)

  • Kim, Dae-Ho;Park, Jin-Hong;Kim, Jung-Hwa;Kim, Cheol-Hee;You, Jin-Hyun;Kwon, Min-Chul;Lee, Hyeon-Yong
    • Korean Journal of Medicinal Crop Science
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    • v.13 no.3
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    • pp.81-86
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    • 2005
  • The immune activities of the extracts from Ephedrae Herba and Rubi Frutus extraction with ultrasonification at $60^{\circ}C$ were compared with the extracts though water extraction at $100^{\circ}C$. The growth of human T cells was increased up to $13{\times}10^4\;viable\;cells/m{\ell}$ in adding $1.0\;g/{\ell}$ of the ultrasonification extracts of R. Fructus at $60^{\circ}C$, compared to adding the extracts at $100^{\circ}C$. The secretion of $TNF-{\alpha}$ for human T cells were also increased up to $13.9{\times}10^{-4}$ pg/cell by adding extracts at $60^{\circ}C$, compared to extracts at $100^{\circ}C$. The extracts of R. Fructus at $60^{\circ}C$ increased NK cell activities up to 50% and the secretion of $NO^{-1}$ from macrophage to $31\;{\mu}M$ for ultrasonification extracts at $60^{\circ}C$.

Selective Expansion of TCR $V{\beta}3$+CD4+T Cells in Collagen-induced Arthritis in DBA/1 Mice (콜라겐 유도 관절염에서 콜라겐 항원 특이 $V{\beta}3$+CD4+T 세포의 선택적 증식)

  • Lee, Jae-Seon;Cho, Mi-La;Lee, Jung-Eun;Min, So-Youn;Yoon, Chong-Hyeon;Kim, Wan-Uk;Min, Jun-Ki;Park, Sung-Hwan;Kim, Ho-Youn
    • IMMUNE NETWORK
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    • v.5 no.2
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    • pp.78-88
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    • 2005
  • Background: Collagen-induced arthritis (CIA) in mice is animal model of autoimmune disease known as rheumatic arthritis in human. We investigated CII-specific CD4+ T cell receptor usage in CIA mice. Methods: In CIA model, draining lymph node (dLN) CD4+ T cells and splenocytes at $3^{rd},\;5^{th},\;8^{th}$ week, we investigated CII-specific T cell proliferation, production of IL-17, IFN-${\gamma}$, TNF-${\alpha}$, IL-4 and IL-10. And we also performed anti-CII IgG Ab measurements in serum level, TCRV ${\beta}$ usage and T cell clonality with RT-PCR-SSCP analysis. Also, we performed proliferative response against CII when CII-specific T cell subset is deleted. Results: CIA mice showed more increase in the serum level of anti-CII IgG than normal mice after induction of arthritis. And the level of anti-CII IgG2a in CIA mice was increased after $3^{rd}$ week after primary immunization, while anti-CII IgG1 was decreased. Draining LN CD4+ T cells have proliferated against CII stimulation at $3^{rd}$ week after $1^{st}$immunization. CD4+T cells derived from dLN of CIA mice produced proinflammatory cytokine IFN-${\gamma}$, IL-17 etc. Draining LN CD4 T cells of CIA presented higher proportion of CD4+V ${\beta}3$+subset compared to those of normal mice at $3^{rd}$ week after $1^{st}$ immunization, and they were increased in proportion by CII stimulation. Draining LN CD4+ T cells without TCRV ${\beta}3+/V{\beta}8.1/8.2+/V{\beta}$10b+cells were not responsive against CII stimulation. But, CII-reactive response of TCRV ${\beta}3-/V{\beta}8.1/8.2-/V{\beta}$10b- T cells was recovered when $V{\beta}3+$ T cells were added in culture. Conclusion: Our results indicate that CD4+$V{\beta}3+$ T cells are selectively expanded in dLN of CIA mice, and their recovery upon CII re-stimulation in vitro, as well as the production Th1-type cytokines, may play pivotal role in CIA pathogenesis.

Effects of Enterococcus faecalis sonicated extracts on IL-2, IL-4 and TGF-β1 production from human lymphocytes (Enterococcus faecalis 추출물이 임파구의 IL-2, IL-4, TGF-β1 분비에 미치는 영향에 관한 연구)

  • Kim, Hyeon-Sik;Lee, Woo-Cheol;Jang, Seok-Woo;Shon, Wan-Jun;Lee, Sang-Takg;Kim, Cheol-Ho;Lim, Sung-Sam
    • Restorative Dentistry and Endodontics
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    • v.30 no.1
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    • pp.1-6
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    • 2005
  • In order to examine the immunoresponse of host cells to Enterococcus faecalis, this in vitro study monitored the production of Interleukin-2 (IL-2), Interleukin-4 (IL-4) and Transforming growth factor-$\beta1\;(TGF-\beta1)$ in human lymphocytes. Lymphocytes were activated with PHA in the presence or abscence of sonicated extracts of E. Faecalis (SEF) and further incubated for 72 hours. The level of each cytokine was measured by ELISA. Data were analyzed with Kruskal-Wallis test and Mann-Whitney U test (P < 0.05). PHA-activated group did exhibit higher level of IL-2 and IL-4 than untreated control group. The levels of expression of both cytokines were significantly decreased following the treatment of high (25 ${\mu}g/ml$) and medium concentration (12.5 ${\mu}g/ml$)) of SEF (P > 0.05) than those of PHA activated group. But low concentration (5 ${\mu}g/ml$)) of SEF showed th similar level of IL-2 and IL-4 production as those of PHA activated group. $TGF-\beta1$ was unaffected by SEF treatment. These results suggested that E. faecalis may suppress IL-2 and IL-4 production by lymphocytes and this could be one of possible factors why E. faecalis are found frequently in the teeth with failed endodontic treatment.

Carbohydrate Metabolism in Preimplantation Stage Embryos and the Role of Metabolites (착상전 초기 배아에서 탄수화물 대사와 그 대사물의 역할)

  • Cheon, Yong-Pil
    • Development and Reproduction
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    • v.12 no.1
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    • pp.19-30
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    • 2008
  • Proper development of fertilized oocyte to blastocyst is a key step in mammalian development to implantation. During development of preimplantation embryos, the mammalian embryo needs supply the energy substrate for keep viability. Usually mammalian oocyte get substrate especially energy substrate from oviduct and uterus, because it does not store much substrate into cytoplasm during oogenesis. Carbohydrates are known as a main energy substrate for preimplantation stage embryos. Glucose, lactate and pyruvate are essential component in preimplantation embryo culture media and there are stage specific preferences to them. Glucose transporter and $H^+$-monocarboxylate cotransporter are a main mediator for carbohydrate transport and those expression levels are primarily under the control of intrinsic or extrinsic factors like insulin and glucose. Other organic substances, amino acids, lipids and nucleotides are used as energy substance and cellular regulation factor. Though since 1960s, successful development of fertilized embryo to blastocyst has been accomplished with chemically defined medium for example BWW and give rise to normal offspring in mammals, the role of metabolites and the regulation of intermediary metabolism are still poorly understood. Glucose may permit expression of metabolic enzymes and transporters in compacting morula, capable of generating the energy required for blastocyst formation. In addition, it has been suggested that the cytokines can modulate the metabolic rate of carbohydrate in embryos and regulate the preimplantation embryonic development through control the metabolic rate. Recently we showed that lactate can be used as a mediator for preimplantation embryonic development. Those observations indicate that metabolites of carbohydrate are required by the early embryo, not only as an energy source, but also as a key substrate for other regulatory and biosynthetic pathways. In addition metabolites of carbohydrate may involve in cellular activity during development of preimplantation embryos. It is suggested that through these regulation and with other regulation mechanisms, embryo and uterus can prepare the embryo implantation and further development, properly.

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Anti-allergic activities of Castanea crenata inner shell extracts fermented by Lactobacillus bifermentans (유산균 발효에 의한 율피(Castanea crenata inner shell) 열수추출물의 아토피 피부 질환에 관한 효과 연구)

  • Choi, Mi-Ok;Kim, Bae-Jin;Jo, Seung-Kyeung;Jung, Hee-Kyoung;Lee, Jin-Tae;Kim, Hak-Yoon;Kweon, Dae-Jun
    • Korean Journal of Food Preservation
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    • v.20 no.4
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    • pp.583-591
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    • 2013
  • Atopic dermatitis (AD) is a common chronic inflammatory disease associated with a cutaneous hypersensitivity reaction to an allergen. Although the incidence of AD is increasing these days, therapeutics has yet to be developed for its treatment. The aim of this study was conducted in order to compare and investigate the characteristic between the Castanea crenata inner shell extract (CS) and the Castanea crenata inner shell extract fermented by Lactobacillus bifermentans (FCS) for an anti-atopic medication. The total polyphenol and flavonoid contents were similar to CS and FCS. In the DPPH and superoxide anion radical scavenging, the CS and FCS had the potential for antioxidant activities. Both of them did not exhibit cytotoxicity to HS68 cells. The evaluation of the anti-inflammatory activity in Raw264.7 cells demonstrated that the FCS has inhibited the LPS-induced production of nitric oxide as compared to the CS. The anti-atopic dermatitis test was done through the induction of DNCB in AD hairless mice. The FCS has inhibited the development of the atopic dermatitis-like skin lesion by transdermal water loss, melanin and erythema of the skin as compared to the CS. Moreover, the pro-inflammatory cytokine IL-$1{\beta}$ and TNF-${\alpha}$ production in hairless mice were inhibited by the FCS treatment. It indicates that the fermentation of the Castanea crenata inner shell has the potential for the treatment of atopic dermatitis.

Anti-inflammation Activities of Cultured Products from Suspension Culture of Aloe vera Callus (Aloe vera Callus 현탁배양 생성물의 항염증 활성)

  • Kim, Myung Uk;Cho, Young Je;Lee, Shin Young
    • Journal of Applied Biological Chemistry
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    • v.56 no.3
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    • pp.157-163
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    • 2013
  • Cultured products (callus and exopolysaccharide) were obtained from suspension culture of Aloe vera callus, and the extracts of callus were further prepared with cold water or 60% ethanol solution. The ethanol extract of callus (AC) and exopolysaccharide (ACP) of 10 mg/mL exhibited the relatively higher suppression activity of 43.2-52.1% against hyaluronidase activity. Thus, their anti-inflammatory effects were further investigated using animal cell (Raw 264.7) in vitro. Though AC shows a slight suppression effect of cell survival rate (97%) using MTT assay in the presence of $400{\mu}g/mL$ AC- dimethyl sulfoxide (DMSO), cell growth promotion was observed in the other samples of lower levels. It indicates that the ethanol extract of Aloe callus rarely affect cell survival rate in the ranges ($200-400{\mu}g/mL$) used in the study. Using Griess reagent, the suppression of NO production by the aloe callus extract was analyzed by measuring the amount of the nitrite produced in Raw 264.7 culture activated by lipopolysaccharide (LPS). As a result, supplementation of AC-distilled water (DW) and AC-DMSO produced higher levels of NO than the positive control LPS. However, the NO suppression effect by ACP-DW was so intense that lower amount ($80-100{\mu}g/mL$) suppressed NO production to the level of the control. The effect was attributed to the expression of the iNOS. Then, Raw 264.7 cells were stimulated with the LPS and expression of COX-2 protein level was analyzed depending on the Aloe suspension culture product treatment. The results showed that the ACP-DW supplemented medium did not express COX-2 by itself, and LPS stimulated COX-2 expression was slightly decreased. On the other hand, realtime-PCR analysis of the expression of inflammatory cytokine showed that IL-$1{\beta}$ and TNF-${\alpha}$ expression was highly suppressed in the ACP- distilled water supplemented medium.