• 한국약용작물학회지
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• 제27권3호
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• pp.218-231
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• 2019

Background: We recently reported that Salvia plebeia R. Br. extracts suppress leukotriene production and effectively inhibit the airway inflammatory response by modulating inflammatory chemokine and cytokine expression. Here, we investigated the synergistic airway anti-inflammation effect of Salvia plebeia and Panax ginseng (Korean red ginseng, KRG) that has been used to treat various immune diseases such as asthma. Methods and Results: To evaluate the synergistic airway anti-inflammatory effect of Salvia plebeia and KRG, we measured the inhibitory effect of monotheraphy with either or co-theraphy with both on leukotriene and reactive oxygen species (ROS) production. Using coal a combustion, fly ash, and diesel exhaust particle (CFD)-induced respiratory disease mouse model, we found that co-theraphy synergistically suppressed airway inflammatory signs such as alveolar wall thickness and collagen fibers deposition, and decreased the number of total cell, $CD11b^+Gr-1^+$ cells, and inflammatory cytokines (IL17A, TNF, MIP-2 and CXCL-1) in bronchoalveolar lavage (BAL) fluid. Conclusions: We confirmed respiratory protection as a therapeutic effect of the Salbia plebeia-KRG 3 : 1 complex (KGC-03-PS) via anti-tracheal muscle contraction and expectorant animal studies using a CFD-induced respiratory disease mouse model.

• Asian Pacific Journal of Cancer Prevention
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• 제15권9호
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• pp.4039-4044
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• 2014

Background: The aim of this study was to investigate the anti-cancer effects and mechanisms of immunochemotherapy of 5-fluorouracil (5-FU) and interleukin-2 (IL-2) on non-small cell lung cancer (NSCLC) A549 cells. Materials and Methods: In order to detect whether 5-FU+IL-2 could effectively inhibit tumor growth in vivo, we established an A549-bearing nude mouse model. The cytotoxicity of natural killer (NK) cells was evaluated using a standard chromium release assay. To evaluate the relevance of NK cells in 5-FU+IL-2-mediated tumor inhibitory effects, we depleted NK cells in A549-bearing mice by injecting anti-asialo-GM-1 antibodies. Effects of 5-FU+IL-2 on the expression and promoter activity of NKG2D ligands (MICA/MICB) in A549 cells in vitro were also assessed. Results: In A549-bearing nude mice, combination therapy significantly inhibited tumor growth in comparison with monotherapy with 5-FU or IL-2 and enhanced the recognition and lysis of tumor cells by NK cells. Further study of mechanisms showed that NK cells played a vital role in the anticancer immune response of 5-FU+IL-2 immunochemotherapy. In addition, the combination therapy synergistically stimulated the expression and promoter activity of MICA/MICB. Conclusions: 5-FU and IL-2 immunochemotherapy significantly inhibited tumor growth and activated NK cytotoxicity in vivo, and these effects were partly impaired after depleting NK cells in tumor-bearing mice. Combination treatment of 5-FU and IL-2 upregulated the expression and the promoter activity of MICA/MICB in A549 cells, which enhanced the recognition of A549 cells by NK cells. All of the data indicated that immunochemotherapy of 5-FU and IL-2 may provide a new treatment option for patients with lung cancer.

• IMMUNE NETWORK
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• 제13권5호
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• pp.213-217
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• 2013

Enterotoxigenic Bacteroides fragilis (ETBF) is a human gut commensal bacteria that causes inflammatory diarrhea and colitis. ETBF also promotes colorectal tumorigenesis in the Min mouse model. The key virulence factor is a secreted metalloprotease called B. fragilis toxin (BFT). BFT induces E-cadherin cleavage, cell rounding, activation of the ${\beta}$-catenin pathway and secretion of IL-8 in colonic epithelial cells. However, the precise mechanism by which these processes occur and how these processes are interrelated is still unclear. E-cadherin form homophilic interactions which tethers adjacent cells. Loss of E-cadherin results in detachment of adjacent cells. Prior studies have suggested that BFT induces IL-8 expression by inducing E-cadherin cleavage; cells that do not express E-cadherin do not secrete IL-8 in response to BFT. In the current study, we found that HT29/C1cells treated with dilute trypsin solution induced E-cadherin degradation and IL-8 secretion, consistent with the hypothesis that E-cadherin cleavage causes IL-8 secretion. However, physical damage to the cell monolayer did not induce IL-8 secretion. We also show that EDTA-mediated disruption of E-cadherin interactions without E-cadherin degradation was sufficient to induce IL-8 secretion. Finally, we determined that HT29/C1 cells treated with LiCl (${\beta}$-catenin activator) induced IL-8 secretion in a dose-dependent and time-dependent manner. Taken together, our results suggest that BFT induced IL-8 secretion may occur by the following process: E-cadherin cleavage, disruption of cellular interactions, activation of the ${\beta}$-catenin pathway and IL-8 expression. However, we further propose that E-cadherin cleavage per se may not be required for BFT induced IL-8 secretion.

• Applied Biological Chemistry
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• 제48권4호
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• pp.411-417
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• 2005

마우스 대식세포주인 RAW264.7 세포에서 곽향(Agastache rugosa)을 포함한 21종의 한약재에서 제조한 열수추출물의 NO생산에 대한 조절효과를 조사하였다. 모든 한약재 추출물은 LPS자극으로 생산된 NO에 대하여 뚜렷한 소거활성을 보이지 않았으나, LPS 무처리 조건에서 곽향이 RAW264.7 세포의 NO생산을 강력하게 유도하였다. $200{\mu}M$의 NOS2의 저해제인 $N^G-monomethyl-L-arginine(N^GMMA$)의 처리에 의하여 곽향이 유도하는 NO 생산은 유의적으로 감소되었다. 또한 $NF-{\kappa}B$ 저해제인 pyrrolidine dithiocarbamate(PDTC)의 처리로 NO 생산이 $100{\mu}M$에서 약 79%까지 감소하였다. 이상의 실험 결과는 곽향 열수추출물이 RAW264.7 세포의 NOS2 발현의 이차적인 세포 내 신호를 발생시킬 수 있으며, NO는 L-arginine 의존적 경로에 의하여 생성된다는 사실을 시사하였다.

• Journal of Acupuncture Research
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• 제24권5호
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• pp.171-183
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• 2007

Objectives : The purpose of this study was to analyze the effects of Erythrinae Cortex herbal-acupuncture solution(EC-HAS) at the Joksamni($ST_{36}$) of mice with collagen II-induced arthritis(CIA). Methods : The author performed several experimental procedures to observe the effects of the EC-HAS at the arthritis. The severity of arthritis, changes of cytokine level and antibody level, histological changes of the CIA mouse joint were analyzed. Results : 1. The incidence of arthritis and arthritis index were significantly decreased in the cases which were treated with the EC-HA. 2. Cartilage destruction and synovial cell proliferation were reduced, and the expression of the collagen fibers was similar with that of the normal group. 3. The levels of IL-6, $IFN-{\gamma}$, $TNF-{\alpha}$, $IL-1{\beta}$ in serum of the CIA mice which were treated with the EC-HA were significantly decreased compared with those of the normal group. 4. The levels of IgG, IgM and anti-collagen II in serum of CIA mice when they treated with the EC-HA were significantly decreased compared with those of the normal group. 5. The expression ratio of $CD4^+$ to $CD8^+$ cells in the EC-HA treated mice were maintained as much as the normal group of the lymph nodes in the CIA mice. 6. The $CD3e^+CD69^+$ and $CD11b^+Gr-1^+$ cell populations in the knee joint were significantly decreased in the EC-HA treated group. Conclusions : These results suggest that the EC-HA at the ST36 may be responsible roles to control on the synovial cell proliferation and to prevent the cartilage destruction in rheumatoid arthritis. These results will be important supporting evidence for the practical use of the EC-HA at rheumatoid arthritis clinic in the future.

• Toxicological Research
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• 제8권2호
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• pp.191-203
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• 1992

The immunosuppressive effects of safrole were studied in female BALB/c mouse. Mice were given 100,200and 400mg safrole/kg daily for 14days and evaluated on day 15. The day 4 immunogloblin-M antibody response to T-dependent antigen, sheep red blood cells (SRBC) was inhibited dose-dependently in all doses studied. In vitro antibody response to polyclonal antigen, lipopolysaccharide (LPS) by spleen cell suspensions from safrole-treated mice were also significantly inhibited. When safrole was treated for 14days to mice, and mitogen-induced proliferation of splenocytes were assayed on day 15, there were significant suppression of responses to B-cell mitogen, LPS and T-cell mitogen concanavalin A(Con A) at a dose of 400mg safrole/kg. Direct addition of safrole on the splenocyte culture also produced a dose dependent suppression on in vitro antibody response to LPS, and mitogen-induced lymphoproliferatin at doses of 100,200,400 and 800${\mu}M$ safrole. The role of metabolic activation in safrole-induced suppression of in vitro antibody response was studied using splenocyte-hepatocyte coculture system. The suppression of in vitro antibody respose to LPS by safrole was not altered when safrole were incubated in the splenocyte-hepatocyte system for 4hr as compared with direct addition of safrole in splenocytes culture. Neither the addition of salicylamide, sulfotransferase inhibitor, nor the addation of inorganic sulfate, sulfation cofactor to the splenocyte-hepatocyte coculture, altered the suppression of antibody response by safrole. These results suggest that the immunosuppression by safrole may not by produced by the reactive metabolites which are mediated in carcinogenesis of safrole.

• 대한수의학회지
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• 제27권1호
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• pp.53-60
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• 1987

This experiment was performed on mice to investigate the effects of an immunopotentiator, picibanil(PC), on the immune responses such as phagocytic activity of reticuloen-dothelial(RE) system, E rosette formation rate of splenic lmphocytes and morphological changes of lymph node tissue. Groups of mice were treated with a single(1KE/kg BW) or sequential(0.1, 0.25 and 0.5KE/kg BW for successive 3 days) intravenous injections of PC. PC treated and untreated control mice were sensitized with 50% sheep erythrocyte suspension(0.2ml/mouse) at 1, 3, 5, 7 and 10 days after PC treatment. Functional and morphological examinations were carried out 5 days after sensitization. The following results were obtained: The phagocytic activity of RE system and the weight of liver and spleen were increased significantly at 3rd, 5th and 7th day. The peripheral polymorphonuclear leukocyte and percent of lymphocyte and monocyte were slightly increased. The rates of E rosette formation of splenic lymphotytes, sequential PC treated groups were more increased at 3rd and 5th day in sequential PC treated groups than in single treated groups. Thereafter it returned gradually to the control level by the time of 10th day. Microscopically primary lymph follicles with indistinct germinal center (GC) were partially disrupted and the parafollicular areas were consisted of the pyroninophilic cells in control group. In PC treated group, the parafollicular areas were markedly proliferated and developments of secondary lymph follicles with enlarged and prominent GC were more pronounced in the sequential injected groups compared to single injected groups. These results indicate that PC affected not only parafollicular area of the T-cell area, but also GC of the B-cell area. It suggests that PC may potentiate both cell mediated immunity and humoral immunity.

• 대한수의학회지
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• 제52권2호
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• pp.89-97
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• 2012

Mycobacterial cell-wall skeleton (CWS) is an immunoactive and biodegradable particulate adjuvant and has been tried to use for immunotherapy. The CWS of Mycobacterium bovis bacillus Calmette-Guerin (BCG-CWS) was studied as an universal vaccine vehicle for antigen conjugation, to develop potentially effective and safe vaccine. Although a variety of biological activities of BCG-CWS have been studied, the effects of BCG-CWS on spleen cells are not fully elucidated. Using MTT assay and trypan blue exclusion test, we found that BCG-CWS significantly enhanced the viability and proliferation of cells. Multiple clusters, indicating proliferation, were observed in BCG-CWS-treated spleen cells and surface marker staining assay revealed that BCG-CWS promoted the proliferation of $CD19^+$ B lymphocyte rather than $CD4^+$ or $CD8^+$ T lymphocyte. In addition, BCG-CWS up-regulated the expression of anti-apoptotic molecules such as bcl-2, bcl-xL. BCG-CWS increased the surface expression of CD25 and CD69 as well as IL-2 production of spleen cells, suggesting increased activation. Furthermore, BCG-CWS enhanced the antigen-specific cell proliferation and interferon-gamma production of spleen cells. Taken together, these results demonstrate the immunostimulatory effects of BCG-CWS on spleen cells via multiple mechanisms, providing valuable information to broaden the use of BCG-CWS in clinical and research settings.

• 대한본초학회지
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• 제26권4호
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• pp.187-194
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• 2011

Objective : Astragali Radix (AR) and Cinnamomi Cortex (CC) are used to enhance immune response in Asian traditional medicine. Immuno-potentiation of the combination of AR and CC were evaluated on the cellular and humoral immune response using murine macrophage cell line (RAW 264.7) and OVA-immunized mice. Methods : This study was designed to investigate the immuno-potentiative effects of AR, CC, and AR with CC on nitric oxide synthesis in RAW 264.7 cells and proliferation and production levels of Intereukin-2 (IL-2) in mouse splenocytes. In addition, we evaluated the plasma-specific antibody responses and splenocyte proliferation on ovalbumin (OVA)-immunized mice treated with herbal extracts. Results : Combination treatment with AR and CC increased nitric oxide synthesis in RAW 264.7 cells and IL-2 level in splenocytes (p<0.001). Combination of AR and CC significantly enhanced the Concanavalin A- (Con A ; T cell mitogen) and lipopolysaccharide-(LPS ; B cell mitogen) induced splenocyte proliferation on the OVA-immunized mice. Combination of AR and CC also significantly enhanced plasma levels of OVA-specific IgG (p<0.01), IgG1 (p<0.05) and total IgM (p<0.01) compared with the OVA-immunized control group. Conclusion : These results suggest that combination of AR and CC could be used as therapeutic profile on activation of immune response.

• Molecules and Cells
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• 제43권7호
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• pp.662-670
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• 2020

We have investigated the involvement of the pre-mRNA processing factor 4B (PRP4) kinase domain in mediating drug resistance. HCT116 cells were treated with curcumin, and apoptosis was assessed based on flow cytometry and the generation of reactive oxygen species (ROS). Cells were then transfected with PRP4 or pre-mRNA-processing-splicing factor 8 (PRP8), and drug resistance was analyzed both in vitro and in vivo. Furthermore, we deleted the kinase domain in PRP4 using Gateway™ technology. Curcumin induced cell death through the production of ROS and decreased the activation of survival signals, but PRP4 overexpression reversed the curcumin-induced oxidative stress and apoptosis. PRP8 failed to reverse the curcumin-induced apoptosis in the HCT116 colon cancer cell line. In xenograft mouse model experiments, curcumin effectively reduced tumour size whereas PRP4 conferred resistance to curcumin, which was evident from increasing tumour size, while PRP8 failed to regulate the curcumin action. PRP4 overexpression altered the morphology, rearranged the actin cytoskeleton, triggered epithelial-mesenchymal transition (EMT), and decreased the invasiveness of HCT116 cells. The loss of E-cadherin, a hallmark of EMT, was observed in HCT116 cells overexpressing PRP4. Moreover, we observed that the EMT-inducing potential of PRP4 was aborted after the deletion of its kinase domain. Collectively, our investigations suggest that the PRP4 kinase domain is responsible for promoting drug resistance to curcumin by inducing EMT. Further evaluation of PRP4-induced inhibition of cell death and PRP4 kinase domain interactions with various other proteins might lead to the development of novel approaches for overcoming drug resistance in patients with colon cancer.