• Title/Summary/Keyword: cytokine mRNA

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Pycnogenol, a Standardized Extract of French Maritime Pine Bark, Inhibited the Transcriptional Expression of Th2 Cytokines by Suppressing $NF-{\kappa}B$ Activation in Primary Splenocytes of C57BL/6 Mice with Murine AIDS (Murine AIDS 감염쥐 splenocytes에서 $NF-{\kappa}B$의 활성화 억제를 통한 pycnogenol의 Th2 cytokines의 mRNA 발현 조절 효과)

  • Lee, Jeong-Min
    • Korean Journal of Food Science and Technology
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    • v.38 no.6
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    • pp.829-834
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    • 2006
  • LP-BM5 murine leukemia retrovirus induces the immune dysfunction by imbalanced secretion of Th1 and Th2 cytokines in the murine AIDS model. In the present study, it was investigated whether pycnogenol (Pyc) administration could deactivate $NF-{\kappa}B$ to regulate the gene expression of Th1 and Th2 cytokines in C57BL/6 mice with murine AIDS. Treatment with Pyc for 12 weeks significantly inhibited the loss of body weight and enlargement of spleen and lymph node usually seen with AIDS. Moreover, Pyc increased the plasma level of Th1 cytokines, IL-2 and $IFN-{\gamma}$, while reducing the plasma level of Th2 cytokines, IL-6, IL-10, and $TNF-{\alpha}$. In primary culture of splenocytes, mRNA expression of Th2 cytokines was suppressed, but that of Th1 cytokines was not affected. The LP-BM5 retrovirus infection stimulated the cytoplasmic activation of $NF-{\kappa}B$ and nuclear translocation of $I-{\kappa}B$, whereas Pyc administration significantly reduced $NF-{\kappa}B$ activation and $I-{\kappa}B$ degradation. These results suggested that the inhibitory effect of Pyc on Th2 cytokines in mice with murine AIDS was dependent on suppression of the $NF-{\kappa}B$ signaling pathway and was not dependent on $INF-{\gamma}$ level, which regulates Th2 cytokines.

Studying of the Effects of Gamichungsangboha-tang water-extract and ethanol-extract on $IFN{\gamma}$, IL-4, T-bet, GATA-3, c-maf in Asthma Model (가미청상보하탕(加味淸上補下湯)의 물과 에탄올 추출법에 따른 항(抗)알레르기 효과(效果)에 대(對)한 연구(硏究))

  • Heo, Tae-Seok;Ko, Eun-Jung;Jung, Hee-Jae;Rhee, Hyung-Koo;Bae, Hyun-Su;Jung, Sung-Ki
    • The Journal of Internal Korean Medicine
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    • v.27 no.1
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    • pp.40-54
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    • 2006
  • Objective: This study aimed to identify the different effects of GMCSBHT water-extract and ethanol-extract on Th1/Th2 differentiation by monitoring Th1/Th2 specific cytokine secretion patterns and the transcriptional activities of T-bet, GATA-3, c-maf, $INF{\gamma}$ and IL-4. Materials and Methods: Spleen cells from eight week-old BALB/c mice were cultured in GMCSBHT extracts containing medium without activation for 24 hours and with activation for 48 hours. CD4+ T cells were isolated and mRNA expression levels of $INF{\gamma}$, IL-4, T-bet, GATA-3, c-maf by RT-PCR and secretion cytokines levels of $IFN{\gamma}$, IL-4 by ELISA were analyzed. Results: GMCSBHT extracts didn't have mitogenic effects on the unstimulated CD4+ T cells. In Th1 skewed condition, GMCSBAHT water extract had no significant effects on mRNA expression levels of $IFN{\gamma}$, T-bet and c-maf, but inhibited mRNA expression levels of IL-4, GATA-3. It showed significantly increased secretion cytokine levels of $IFN{\gamma}$, but had no significant effect on secretion cytokine levels of IL-4. In Th2 skewed condition, GMCSBHT ethanol extract inhibited mRNA expression levels of $INF{\gamma}$, IL-4, GATA-3 and c-maf significantly, but had no significant effects on mRNA expression levels of T-bet. It had no significant effects on secretion cytokine levels of $INF{\gamma}$, but showed remarkable inhibitory effects on secretion cytokine levels of IL-4. Conclusion: Results suggest that on Th1/Th2 deviation, GMCSBHT water extract has both amplifying effects on Th1 differentiation and inhibitory effects on Th2, but GMCSBHT ethanol extract has stronger inhibitory effects on Th2 differentiation than on Th1.

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Changes of Cytokine and Chemokine mRNA Expression in Whole Blood Cells from Active Pulmonary Tuberculosis Patients after T-Cell Mitogen and Mycobacterium tuberculosis Specific Antigen Stimulation

  • Kim, Sunghyun;Park, Sangjung;Lee, Hyeyoung
    • Biomedical Science Letters
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    • v.20 no.3
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    • pp.162-167
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    • 2014
  • Tuberculosis (TB) is one of the major global health problems and it has been estimated that in 5~10% of Mycobacterium tuberculosis (MTB)-infected individuals, the infection progresses to an active disease. Numerous cytokines and chemokines regulate immunological responses at cellular level including stimulation and recruitment of wide range of cells in immunity and inflammation. In the present study, the mRNA expression levels of eight host immune markers containing of IFN-${\gamma}$, TNF-${\alpha}$, IL-2R, IL-4, IL-10, CXCL9, CXCL10, and CXCL11 in whole blood cells from active pulmonary TB patients were measured after T-cell mitogen (PHA) and MTB specific antigens (ESAT-6, CFP-10, and TB7.7). Among the TH1-type factors, IFN-${\gamma}$ mRNA expression was peaked at 4 h, TNF-${\alpha}$ and IL-2R mRNA expression was significantly high at the late time points (24 h) in active TB patients, TH2-type cytokine (IL4 and IL10) mRNA expression levels in both active TB and healthy controls samples did not changed significantly, and the mRNA expression of the three IFN-${\gamma}$-induced chemokines (CXCL9, CXCL10, and CXCL11) were peaked at the late time points (24 h) in active TB patients after MTB specific antigen stimulation. In conclusion, the mRNA expression patterns of the TB-related immune markers in response to the T-cell mitogen (PHA) differed from those in response to MTB specific antigens and these findings may helpful for understanding the relationship between MTB infection and host immune markers in a transcripts level.

Expression on Echinostoma hortense Infection Manifestation on the Cytokine of the Splenocytes of BALB/c and C3H/HeN Mice

  • Ryang Yong-Suk;Cho Yoon-Kyung;IM Jee-Aee;Lee Kyu-Je;Kim Insik;Sung Ho-Joong
    • Biomedical Science Letters
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    • v.10 no.3
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    • pp.219-230
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    • 2004
  • This experiment was performed to examine the in vitro and in vivo affects of the two different haplotype strains of mice, BALB/c and C3H/HeN infected with Echinostoma hortense, and the manifestation of the profiles of cytokine in the splenocytes. In the in vitro experiment, the two mice's splenocytes were divided and stimulated with antigen of crude extracts and the antigen of excretory and secretory products of an adult warm and the manifestation of cytokine mRNA was verified with RT-PCR. As a result, the two different strains of mice both strongly manifested the Th2 cytokine rather than the Thl cytokine and in the case of the Th2 cytokine, the BALB/c mice manifested more strongly than the C3H/HeN mice. In the experiment using the ELISA method, the protem cytokine manifestation had the same result as the mRNA experiment. In the in vivo experiment, the mice was infected via oral route with the metacercaria of the Echinostoma hortense and the manifestation of cytokine was verified by RT-PCR and ELISA and the results were the same as the in vitro experiment. Therefore, in the two strains of BALB/c and C3H/HeN, the C3H/HeN showed a higher susceptivity to the Echinostoma hortense.

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Effect of Gamikyejakjimo-tang(jiāwèiguìsháozhīm-tāng) ethanol extract on Rheumatoid Arthritis factors (가미계작지모탕(加味桂芍知母湯) 주정 추출물이 류마티스 관절염 인자에 미치는 영향)

  • Sim, Boo-Yong;Bak, Ji-Won;Kim, Dong Hee
    • The Korea Journal of Herbology
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    • v.29 no.5
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    • pp.65-73
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    • 2014
  • Objectives : The purpose of this study is to prove the effect of Gamikyejakjimo-tang (ji$\bar{a}$w$\grave{e}$igu$\grave{i}$sh$\acute{a}$ozh$\bar{i}$m-t$\bar{a}$ng, GK) on rheumatoid arthritis. Methods : We checked viability and measured production of IL-$1{\beta}$, IL-6, IL-17, TNF-${\alpha}$ in RAW 264.7 cell after treat by GK. Then we measured rheumatoid arthritis index score of DBA/1 mice with rheumatoid arthritis induced by CIA after GK oral administration, checked IL-$1{\beta}$, IL-6, IL-17, TNF-${\alpha}$ and hs-CRP tests in serum. Also we were observed mRNA expression of IL-$1{\beta}$, IL-6, IL-17 and TNF-${\alpha}$ in spleen by RT-PCR. Results : GK showed cell viability of 100% or higher in all concentration in RAW 264.7 cells. GK inhibited LPS-induced productions of rheumatoid arthritis mediators cytokine in RAW 264.7cells. GK treated group showed improvement from rheumatoid arthritis at decreased the index score. Also, GK treated group decreased level in serum of IL-1b, IL-6, IL-17, TNF-a and hs-CRP tests by 31%, 35%, 20%, 57% and 58% respectively. Finally, GK treated group showed decrease expression of IL-$1{\beta}$, IL-6, IL-17 and TNF-${\alpha}$ mRNA in spleen by 46%, 51%, 25% and 42% respectively. Conclusions : In this study, in-vitro and in-vivo results observed rheumatoid arthritis factors cytokine of IL-$1{\beta}$, IL-6, IL-17 and TNF-${\alpha}$ decrease in RAW 264.7 cells, serum, mRNA expression. Also, GK showed decrease of inflammation figure in hs-CRP tests depending on effect of rheumatoid arthritis. Thus, these results can used as a effective drug of GK for rheumatoid arthritis.

A Study on the Effect of Changchuldointanggami-bang(Cāngzhútáoréntāngjiāwèi-fāng) in Rheumatoid Arthritis (창출도인탕가미방(蒼朮桃仁湯加味方)이 류마토이드 관절염에 미치는 실험적 연구)

  • Park, Eun-Sook;Yoon, Il-Ji
    • Journal of Korean Medicine Rehabilitation
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    • v.19 no.4
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    • pp.1-18
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    • 2009
  • Objectives : This study was carried out to understand the immune responses of the Changchuldointanggami-bang(after referred to CDIT) on antioxidants, THP-1 cell and rheumatoid arthritis in Collagen-induced Arthritis(CIA) mice. Methods : The antioxidative effect of CDIT on DPPH free radical and SOD-like activity was measured. CDIT was administered to THP-1 cell, cytokine and mRNA associated with inflammation were measured. CDIT was orally administered to mice with arthritis by collagen II and then cytokine, PBMC in the serum were measured. Results : 1. The scavenging activity of CDIT on DPPH free radical was dose-dependent. 2. The effect of CDIT on SOD-like activity was dose-dependent. 3. The production of $IL-1{\beta}$, IL-6, $TNF-{\alpha}$ was decreased significantly in THP-1 cell. 4. The expression of $IL-1{\beta}$ mRNA, IL-6 mRNA, $TNF-{\alpha}$ mRNA were decreased significantly in THP-1 cell. 5. $IL-1{\beta}$, IL-6, $TNF-{\alpha}$ were decreased significantly in the serum of CIA mice. 6. CD3+, CD4+, CD8+ were increased significantly, but CD3+/CD69+, CD3+/CD49b+, B220+/CD23+ were decreased significantly in PBMC of CIA mice. Conclusions : Taking all these observations into account, CDIT is considered to be effective in rheumatoid arthritis. Therefore we have to survey continuously in looking for the effective substance and mechanism in the future.

The effect of Injinchunggan-tang(Yinchenqinggan-tang) on Inflammatory Cytokine Gene Expression in Kupffer Cells (인진청간탕(茵蔯淸肝湯)이 kupffer cell 의 inflammatory cytokine 발현에 미치는 영향)

  • Kim Ji-Kwon;Kim Young-Chul;Lee Jang-Hoon;Woo Hong-Jung
    • The Journal of Internal Korean Medicine
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    • v.25 no.1
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    • pp.46-58
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    • 2004
  • Objectives : This study was designed to investigate the effects of Injinchunggan-tang(Yinchenqinggan-tang) on the expression of inflammatory cytokine genes and proteins in kupffer cells. Materials and Methods : The mRNA expression level and protein secretion level were measured using quantitative RT-PCR and ELISA assay respectively in Injinchunggan-tang-treated and untreated kupffer cells after exposed to ethanol, acetaldehyde and lipopolysaccharide. Results : Injinchunggan-tang(Yinchenqinggan-tang) reduced mRNA expression level and protein secretion level of $TNF-{\alpha},\;TGF-{\beta}1,\;IL-1{\beta},\;IL-6,\;IL-8$ that are induced by ethanol, acetaldehyde and lipopolysaccharide in kupffer cells and that mediate inflammation and fibrosis of liver. Conclusion : The result indicates that Injinchunggan-tang (Yinchenqinggan-tang) blocks alcohol-induced liver injury and protects liver by reducing production of inflammatory cytokines.

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Polarization of THP-1-Derived Macrophage by Magnesium and MAGT1 Inhibition in Wound Healing

  • Mun Ho Oh;JaeHyuk Jang;Jong Hun Lee
    • Archives of Plastic Surgery
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    • v.50 no.4
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    • pp.432-442
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    • 2023
  • Background Macrophages play a major role in wound healing and prevent infection from the outside. Polarization conversion of macrophages regulates aspects of inflammation, and two macrophages, M1 (classically activated) and M2 (alternatively activated), exist at both ends of broad-spectrum macrophage polarization. Thus, we aimed to investigate whether macrophage polarization can be artificially regulated. To this end, MgSO4 and small-interfering RNA (siRNA) targeting magnesium transport 1 (MAGT1) were used to investigate the effects of intracellular magnesium (Mg2+) concentrations on the differentiation of macrophages in vitro. Methods THP-1 derived macrophages maintained in a culture medium containing 5 mM MgSO4 and siRNA to inhibit the expression of MAGT1. As comparative groups, THP-1 derived macrophages polarized into M1 and M2 macrophages by treatment with M1, M2 inducer cytokine. The polarization status of each group of cells was confirmed by cell surface antigen expression and cytokine secretion. Results We found that MgSO4 treatment increased CD163 and CD206, similar to the effect noted in the M2 group. The expression of CD80 and HLA-DR was increased in the group treated with MAGT1 siRNA, similar to the effect noted in the M1 group. Functional assays demonstrated that the group treated with MgSO4 secreted higher levels of IL-10, whereas the MAGT1 siRNA-treated group secreted higher levels of IL-6 cytokines. Additionally, the conditional medium of the Mg2+ treated group showed enhanced migration of keratinocytes and fibroblasts. Conclusion Mg2+ can help to end the delay in wound healing caused by persistent inflammation in the early stages.

Effect of NUCKS-1 Overexpression on Cytokine Profiling in Obese Women with Breast Cancer

  • Soliman, Nema Ali;Zineldeen, Doaa Hussein;El-Khadrawy, Osama Helmy
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.2
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    • pp.837-845
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    • 2014
  • Background: Overweight and obesity are recognized as major drivers of cancers including breast cancer. Several cytokines, including interleukin-6 (IL-6), IL-10 and lipocalin 2 (LCN2), as well as dysregulated cell cycle proteins are implicated in breast carcinogenesis. The nuclear, casein kinase and cyclin-dependent kinase substrate-1 (NUCKS-1), is a nuclear DNA-binding protein that has been implicated in several human cancers, including breast cancer. Objectives: The present study was conducted to evaluate NUCKS-1 mRNA expression in breast tissue from obese patients with and without breast cancer and lean controls. NUCKS-1 expression was correlated to cytokine profiles as prognostic and monitoring tools for breast cancer, providing a molecular basis for a causal link between obesity and risk. Materials and Methods: This study included 39 females with breast cancer (G III) that was furtherly subdivided into two subgroups according to cancer grading (G IIIa and G IIIb) and 10 control obese females (G II) in addition to 10 age-matched healthy lean controls (G I). NUCKS-1 expression was studied in breast tissue biopsies by means of real-time PCR (RT-PCR). Serum cytokine profiles were determined by immunoassay. Lipid profiles and glycemic status as well as anthropometric measures were also recorded for all participants. Results: IL-6, IL-12 and LCN2 were significantly higher in control obese and breast cancer group than their relevant lean controls (p<0.05), while NUCKS-1 mRNA expression was significantly higher in the breast cancer group compared to the other groups (p<0.05). Significant higher levels of IL-6, IL-12, and LCN2 as well as NUCKS-1 mRNA levels were reported in G IIIb than G IIIa, and positively correlated with obesity markers in all obese patients. Conclusions: Evaluation of cytokine levels as well as related gene expression may provide a new tool for understanding interactions for three axes of carcinogenesis, innate immunity, inflammation and cell cycling, and hope for new strategies of management.

Changes of cytokine mRNA expression and IgG responses In rats Infected with Capillaria hepatica

  • Kim, Dong-Kwan;Joo, Kyoung-Hwan;Chung, Myung-Sook
    • Parasites, Hosts and Diseases
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    • v.45 no.2 s.142
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    • pp.95-102
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    • 2007
  • The mRNA expression of several cytokines was evaluated in splenocytes and mesenteric lymph node (MLN) cells of rats infected with Capillaria hepatica by reverse-transcription (RT)-PCR until week 12 after infection. IgG1 and IgG2a, which are associated with Th1 and Th2 response, respectively, were also assessed by ELISA. The results indicated that the majority of cytokines, including the Th1 (IL-2 and $IFN-{\gamma}$ and Th2 cytokines (IL-4, IL-5 and IL-10) were expressed at maximal levels during the early stage of infection (after week 1-2), and the ELISA data also evidenced a similar pattern of changes in IgG1 arid IgG2a. Th1 and Th2 cytokines responded in a similar fashion in this rat model. The expression of cytokines in splenocytes was significantly higher than that in MLN cells, thereby indicating that cytokine production is controlled more by spleen than by MLN. In addition, the observation that $IFN-{\gamma}$ expression increased unexpectedly at the time of maximal egg production (6 weeks after infection) indicated that $IFN-{\gamma}$ is a cytokine reacting against egg production. However, increased IL-5 expression occurring in tandem with worm activity indicated that the activity of C. hepatica might be controlled by IL-5 expression.