• IMMUNE NETWORK
• /
• 제1권3호
• /
• pp.213-220
• /
• 2001

Background: A previous study has shown that Euonymus alatus (EA) has an antidotic activities against inflammation, suggesting possibility that EA can exert this beneficial effects to liver injury by an initial protection against drug-induced hepatocyte demage. The present study was undertaken to evaluate the protective effect of EA-extract on experimentally induced hepatitis in ICR mice and to investigate some mechanisms responsible for its action. Methods: Water EA extract was used in this experiments. The mice received i.p. a dose of 700 mg/kg galactosamine (GalN) together with $5{\mu}g/kg$ of endotoxin (LPS), or received i.v. 12 mg/kg of concanavalin A (Con A). EA (4 mg/mouse) was administrated on day -2, -1 and 0 before induction of liver injury. Liver injury was assessed by measurement of serum alanin amino-transferase (SGPT) levels on 9 hr after GaIN.LPS, or 8 hr after con A administration. Results: Treatment with either GaIN or LPS alone did not cause hepatitis. However, simultaneous administration of GalN and LPS to mice resulted in LPS-dose dependent fulminant hepatitis. GaLN/LPS-induced liver injury was reduced when mice were given EA for 3 days before induction. This preventive effect of Ea was more prominent when EA was given by intraperitoneal route rather then by oral route. Pretreatment of EA or dexamethasone inhibited significantly $TNF{\alpha}$ production in GalL/LPS-injured mice. However, EA-treatment did not influence $TNF{\alpha}$-induced hepatitis in GalN-sensitized mice, suggesting that $TNF{\alpha}$ is likely to act as one of final mediators of endotoxin action and the protective effect of EA might be manifested chiefly by inhibition of endotoxin-induced $TNF{\alpha}$ production, not by blocking the $TNF{\alpha}$-action. Injection of Con A into mice evoked remarkable liver injury in a dose dependent fashion. This liver damage was reduced by EA-pretreatment. Dexamethasone significantly reduced both GalL/LPS-induced and Con A-induced liver damages, showing synergism with EA. However, indomethacin reduced only GalN/ LPS-induced hepatitis, not for Con A-induced hepatitis. Conclusion: These results led to the conclusion that EA may be able to contribute at least in part to prevent the drug-induced hepatotoxicity, and that its anti-hepatitis effects might be manifested directly by modulation of endogenous mediators, such as leukotriese D4, $TNF{\alpha}$ and free radical, and indirectly by regulation of immune mediated responses. Also these results suggested that EA could be developed as a potential antidotic agent.

• Parasites, Hosts and Diseases
• /
• 제30권2호
• /
• pp.125-132
• /
• 1992

Toxoplasma gondii이 약독주인 Beverley주, Fukaya주 띤 MR49주를 각각 감염시킨 BALB/c 마우스에서 발현되는 세포매개 성 및 체액성 면역반응을 비교 관찰할 목적으로 매주 1회씩 10주에 걸쳐 각각 3마리씩 취하여, 소정의 과정을 거쳐 준비한 비장세포에 concanavalin A(Con. A), lipopolysaccharide(LPS) 및 Toxoplasma Iysate를 처리한 후 아세포화 반응의 정도를[3H]-thymidine의 전입량으로 측정, 평가하는 한편, 혈청내 IgG 및 IgM 항체가를 효소결합면역흡착법 (ELISA)으로 측정, 분석하였다. Con. A 및 Toxoplasma Iysate로 처리시 비장세포의 아세포화 반응은 3개주 감염군 모두 대조군에 비해 감염후 1주부터 유의하게 감소되었으나, LPS로 처리시 3개 주 감염군 모두 대조군에 비해 유의한 차이가 없었다 또한 각 주별 감염 마우스 상호간에도 유의 한 차이가 없었다. 혈청내 IgG 항체가는 3개주 감염군 모두 감염후 2주부터 7주까지 계속적으로 증가한 후 그 항체가가 지속되었으며, IgM 항체가는 1주에서 4주 사이에 현저히 높았다. 그러나 카 주별 감염 마우스의 혈청내 IgG 및 IgM 항체가 상호간에는 유의한 차이가 없었다. 이상의 성적으로 보아 T. gondii의 Beverley주, Fukaya주 및 ME49주를 각각 감염시킨 마우스에 있어 감염 초기에는 세포매개성 면역반응이 억제되며, 체액성 면역반응에 있어 IgM 항체는 감염 초기에 증가하였고 IgG 항체는 초기부터 높은 수준으로 장기간 지속됨을 알 수 있었다.

• Journal of the Korean Wood Science and Technology
• /
• 제41권6호
• /
• pp.551-558
• /
• 2013

본 연구는 만병초와 구상나무 추출물의 항알러지 효과에 대해 조사하였다. 비만세포 RBL-2H3에 concanavalin A (Con A)를 처리하여 면역반응을 유도한 후 만병초와 구상나무 추출물을 농도별로 처리하여 real-time PCR을 이용하여 IL-4, IL-13의 mRNA 발현량을 확인하였다. 또한 ${\beta}$-hexosaminidase 분비를 측정하여 탈과립 유도효과를 평가하였다. IL-4의 발현은 만병초 $10^{-7}$, $10^{-5}$, $10^{-3}%$ 농도와 구상나무의 모든 농도에서 유의성 있게 감소하였으며, IL-13의 발현은 만병초와 구상나무 모든 농도에서 유의성 있게 감소되었다. 만병초 추출물과 구상나무 추출물의 $10^{-5}$, $10^{-3}%$ 농도에서는 ${\beta}$-hexosaminidase 분비가 유의성 있게 감소하였다. 이러한 결과를 통해 만병초와 구상나무 추출물이 면역반응 표적유전자의 발현을 감소시키고 면역반응에 의해 유도되는 탈과립유도 효과도 감소시킴을 확인할 수 있었다. 이는 만병초와 구상나무 추출물이 항알러지 효과가 있음을 제시하는 결과이다.

• 대한약리학회지
• /
• 제27권2호
• /
• pp.135-144
• /
• 1991

흰쥐 말초혈액에서 얻은 T 림프구를 아드레날린성 ${\beta}-$수용체 효현제 및 concanavalin A(Con-A)로 자극해 다음과 같은 결과를 얻었다. 자극이 없는 상태에서의 주 인산화 단백은 분자량 44kD, 등전점 6.8의 단백이었으며 효현제로 자극시키면 분자량 44kD, 등전점 6.3의 단백이 새로이 인산화되어 나타났다. 이 분자량 44kD, 등전점 6.3의 단백은 forskolin에 의해 역시 인산화되며 A-kinase 억제제인 H-8을 전처치하면 인산화의 억제가 나타났다. 또한 Con-A로 자극시키면 44 kD/pI 6.3 단백의 인산화가 증가되었으며 이 인산화의 증가는 CaM kinase 억제제인 W-7 전처치에 의해 억제되었다. H-7은 분자량 44 kD, 등전점 6.8 단백의 인산화를 감소 시켰다. 이상의 결과로 분자량 44 kD 등전점 6.3의 단백은 A-kinase와 CaM kinase 모두에 의해 인산화 되는 기질단백으로서 tryptic peptide map상에서 44 kD/pI 6.8 단백과 44 kD/pI 6.3 단백은 서로 다른 단백임을 알 수 있었다.

• 대한한의학회지
• /
• 제32권3호
• /
• pp.25-34
• /
• 2011

Objectives: We performed simultaneous determination of five constituents by HPLC in Samul-tang (SMT). Additionally, we investigated the immune-stimulatory potential of SMT on specific cellular and humoral immune responses in ovalbumin (OVA)-immunized mice. Methods: Reverse-phase chromatography using a Gemini C18 column operating at $40^{\circ}C$, and photodiode array (PDA) detection at 190-400 nm, were used for quantification of the five components of SMT. Mobile phase using a gradient flow consisted of two solvent systems. Solvent A was 1.0% (v/v) aqueous acetic acid and solvent B was acetonitrile with 1.0% (v/v) acetic acid. C57BL/6 mice were immunized intraperitoneally with OVA/alum ($100{\mu}g/200{\mu}g$) on days 1, 8, and 15. The extract of SMT (1000 mg/kg) was given to mice orally for 21 days (from day 1 to day 21). At day 22, OVA-, lipopolysaccharide (LPS)- and concanavalin A (Con A)-stimulated splenocyte proliferation and OVA-specific and total antibodies were measured in plasma. Results: Calibration curves were acquired with $r^2$>0.9999, and the relative standard deviation (RSD, %) for intra- and inter-day precision were both less than 3.5%. The recovery was in the range of 95.69-115.12%, with an RSD less than 6.0%. The contents of five components in SMT were 1.08-15.30 mg/g. SMT significantly enhanced Con A-induced splenocyte proliferation in OVA-immunized mice (p<0.01). Also, SMT significantly enhanced OVAspecific IgG, IgG1 and total IgM levels in plasma compared with the OVA-immunized group. Conclusions: The established method will be applied for the quantification of major components and immunestimulating activity in OVA-immunized mouse model of SMT.

• Journal of Veterinary Science
• /
• 제25권1호
• /
• pp.1.1-1.15
• /
• 2024

Background: Axitinib, a potent and selective inhibitor of vascular endothelial growth factor (VEGF) receptor (VEGFR) tyrosine kinase 1,2 and 3, is used in chemotherapy because it inhibits tumor angiogenesis by blocking the VEGF/VEGFR pathway. In veterinary medicine, attempts have been made to apply tyrosine kinase inhibitors with anti-angiogenic effects to tumor patients, but there are no studies on axitinib in canine mammary gland tumors (MGTs). Objectives: This study aimed to confirm the antitumor activity of axitinib in canine mammary gland cell lines. Methods: We treated canine MGT cell lines (CIPp and CIPm) with axitinib and conducted CCK, wound healing, apoptosis, and cell cycle assays. Additionally, we evaluated the expression levels of angiogenesis-associated factors, including VEGFs, PDGF-A, FGF-2, and TGF-β1, using quantitative real-time polymerase chain reaction. Furthermore, we collected canine peripheral blood mononuclear cells (PBMCs), activated them with concanavalin A (ConA) and lipopolysaccharide (LPS), and then treated them with axitinib to investigate changes in viability. Results: When axitinib was administered to CIPp and CIPm, cell viability significantly decreased at 24, 48, and 72 h (p < 0.001), and migration was markedly reduced (6 h, p < 0.05; 12 h, p < 0.005). The apoptosis rate significantly increased (p < 0.01), and the G2/M phase ratio showed a significant increase (p < 0.001). Additionally, there was no significant change in the viability of canine PBMCs treated with LPS and ConA. Conclusion: In this study, we confirmed the antitumor activity of axitinib against canine MGT cell lines. Accordingly, we suggest that axitinib can be applied as a new treatment for patients with canine MGTs.

• 한국식품영양과학회지
• /
• 제45권2호
• /
• pp.174-180
• /
• 2016

본 연구에서는 산양유 발효유(F-GM)의 급여가 면역 활성과 체력증진에 미치는 영향을 조사하였다. F-GM을 경구 투여한 마우스의 비장세포는 림프구 유사분열 촉진인자인 ConA와 LPS에 대한 증식반응의 상승을 나타내었다. ConA를 처리한 비장세포로부터 분비되는 cytokine을 ELISA법에 의해 정량한 결과 F-GM 투여에 의해 IL-2와 IFN-${\gamma}$의 생성은 유의하게 증가하였으나, IL-4 및 IL-10의 생성은 별다른 변화를 보이지 않았다. 또한 RT-PCR을 이용한 이들 cytokine mRNA의 정량실험에서도 F-GM 투여로 IL-2와 IFN-${\gamma}$의 mRNA가 증가하는 것을 확인하였다. 한편 KLH(20 mg/mouse)를 정상 대조군 마우스 혹은 F-GM을 급이한 마우스에 각각 면역하고 5주째에 KLH에 대한 항체가를 측정한 결과 F-GM을 급이한 마우스에서 항체가가 유의하게 상승하는 것으로 나타났다. KLH에 대한 항체의 isotype을 조사한 실험에서는 KLH에 대한 IgG1, IgG2a 그리고 IgM 특이항체의 상승이 관찰되었다. F-GM 경구 투여가 세포성 면역에 미치는 영향을 측정하기 위하여 면역개시 7주째의 마우스를 이용하여 KLH에 대한 지연형 과민반응(DTH)을 측정하였다. 그 결과 F-GM을 경구 투여한 마우스에서 DTH의 유의한 상승효과가 확인되었다. 한편 수영시험을 통해 F-GM의 경구 투여가 체력증강에 미치는 영향을 측정한 결과 F-GM을 투여한 마우스에서 매우 유의한 수영시간의 연장 효과가 관찰되었다. 이들 결과로부터 F-GM은 면역세포의 활성화를 통한 면역력 강화는 물론 체력을 증가시키는 생리활성을 갖는 것으로 확인되었다.

• Food Science and Biotechnology
• /
• 제18권6호
• /
• pp.1316-1321
• /
• 2009

Multiple beneficial properties of Opuntia ficus indica (OPF) are well established. In the present study, we have investigated the immunological role of OPF extract (OPFE) on murine splenocytes. OPFE dose- and time-dependently enhanced the proliferation of splenocytes without cytotoxicity. Our results also showed that the number of $CD4^+$ helper T cells and CD45R/$B220^+$ pan B cells increased markedly, but not $CD8^+$ cytotoxic T cells or $CD11b^+$ granulocytes/macrophages. In addition, OPFE significantly decreased the production levels of T helper (Th) 1 type cytokines, interferon (IFN)-$\gamma$, and tumor necrosis factor (TNF)-$\alpha$, although had no significantly differences in those of interleukin (IL)-4, a Th2 type cytokine in concanavalin A (Con A)-stimulated blastogenic cells. Furthermore, OPFE alone strongly increased IL-4 production and decreased TNF-$\alpha$ production even in the absence of Con A. On the basis of these results, this study suggests that OPFE enhances immunity by regulating the pro- and anti-inflammatory response, indicating that this extract exerts a marked immunomodulatory effect, confirming its usefulness as therapy for immune-related diseases.

• 대한한방내과학회지
• /
• 제30권1호
• /
• pp.9-23
• /
• 2009

Objectives : Asthma is a chronic inflammatory disorder of the airways by many cells such as mast cells, Th2 lymphocytes and eosinophile. The present study was aimed to evaluate the effects of Salviae miltiorrhizae (SM) on T cell cytokine production, mast cells. and eosinophils, Methods : We screened 13 herbs to find compounds with potential to control Th cytokine production. using concanavalin A (con A)-activated splenocyte cultures. Con A-activated $IFN-\gamma$ and IL-4 levels in supernatants of splenocyte cultures. Bone marrow derived mast cells (BMMC) were incubated with SM and then the expressions of membrane proteins of BMMC were analyzed by fluorescence activated cell sorter (FACS). BALB/c mice sensitized to ovalbumin (OVA) were challenged with aerosolized OVA for 6 weeks. During the last weeks some mice were treated with SM. Then eosinophils in bronchoalveolar lavage fluid (BALf) were counted and pathologic changes of lung tissue were observed with hematoxylin-eosin stain. Results : SM increased $IFN-\gamma$ level on splenocyte culture significantly. but had no significant effects on expressions of ICAM-1, CD62L, integrin $a_4$. c-kit, IL-3 receptors. CD11a, or IgE receptors of BMMC. SM treatment significantly inhibited eosinophil infiltrates in BALf and peribronchial lung inflammation. Conculusions : The present data suggested that SM may have an effect on Th cytokine secretion and eosinophils associated with asthma responses. Therefore SM might be of therapeutic value in treating asthma.

• Asian-Australasian Journal of Animal Sciences
• /
• 제20권6호
• /
• pp.954-961
• /
• 2007

The experiments were conducted to determine effects of a mixture of conjugated linoleic acid (CLA) isomers on T cell subpopulations and responsiveness to mitogen of splenocytes in male broiler chicks. In experiment 1, birds (8-d old) were fed basal, CLA-(CLA) and safflower oil-supplemented (SA) diets which were formulated by supplementary 10 g CLA or safflower oil/kg to the basal diet for 14 d. Broiler starter diet, which mainly consisted of corn and soybean meal, was served as the basal diet. Proliferative response and interleukin (IL)-2-like activity stimulated by concanavalin (Con) A at a concentration of $10{\mu}g/ml$ of splenocytes in chicks fed the CLA diet were greater than in chicks fed the SA diet, but not at $20{\mu}g$ Con A/ml. Percentage of CD3-positive T cells in splenocytes did not differ between chicks fed the SA diet and CLA. Ratio of CD4-positive T cells to CD8- positive T cells was significantly affected by dietary fat source. In experiment 2, broiler chicks (1-d old) were fed the same diets as in experiment 1 for 14 d. Results of splenocyte proliferation to Con A were similar to those in experiment 1, but phytohemaggulutinin (PHA)- or pokeweed mitogen (PWM)- induced splenocyte proliferation did not differ between the CLA and SA fed groups. Supplementation with SA or CLA to the basal diet tended to have a depressive effect on the proliferation, with the greater effect being that of SA. In experiment 3, effect of an addition of CLA to splenocyte culture medium on splenocyte proliferation was determined. An addition of CLA to the culture medium resulted in reduction of the splenocyte proliferation to Con A, but an addition of linoleic acid. When PWM and PHA were used as mitogen, the inhibitory effect of CLA and linoleic acid on the proliferation did not differ. The results suggested that the effect of dietary CLA on splenocyte proliferation was similar to that of SA, although the effect of dietary CLA on sub-populations was slightly different from that of dietary SA. Further studies are needed to clarify whether use of CLA would be beneficial for maintaining or enhancing T cell immunity in chicks.