• Parasites, Hosts and Diseases
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• 제49권2호
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• pp.115-123
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• 2011

Toxoplasma gondii Korean isolate (KI-1) tachyzoites were inoculated intraduodenally to BALB/c mice using a silicon tube, and the course of infection and immune responses of mice were studied. Whereas control mice, that were infected intraperitoneally, died within day 7 post-infection (PI), the intraduodenally infected mice survived until day 9 PI (infection with $1{\times}10^5$ tachyzoites) or day 11 PI (with $1{\times}10^6$ tachyzoites). Based on histopathologic (Giemsa stain) and PCR (B1 gene) studies, it was suggested that tachyzoites, after entering the small intestine, invaded into endothelial cells, divided there, and propagated to other organs. PCR appeared to be more sensitive than histopathology to detect infected organs and tissues. The organisms spread over multiple organs by day 6 PI. However, proliferative responses of splenocytes and mesenteric lymph node (MLN) cells in response to con A or Toxoplasma lysate antigen decreased significantly, suggesting immunosuppression. Splenic $CD4^+$ and $CD8^+$ T-Iymphocytes showed decreases in number until day 9 PI, whereas IFN-${\gamma}$ and IL-10 decreased slightly at day 6 PI and returned to normal levels by day 9 PI. No TNF-${\alpha}$ was detected throughout the experimental period. The results showed that intraduodenal infection with KI-1 tachyzoites was successful but did not elicit significant mucosal immunity in mice and allowed dissemination of T. gondii organisms to systemic organs. The immunosuppression of mice included reduced lymphoproliferative responses to splenocytes and MLN cells to mitogen and low production of cytokines, such as IFN-${\gamma}$, TNF-${\alpha}$, and IL-10, in response to T. gondii infection.

• Biomolecules & Therapeutics
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• 제31권3호
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• pp.350-358
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• 2023

Hand-foot-and-mouth disease (HFMD) is a viral infectious disease that occurs in children under 5 years of age. Its main causes are coxsackievirus (CV) and enterovirus (EV). Since there are no efficient therapeutics for HFMD, vaccines are effective in preventing the disease. To develop broad coverage against CV and EV, the development of a bivalent vaccine form is needed. The Mongolian gerbil is an efficient and suitable animal model of EV71 C4a and CVA16 infection used to investigate vaccine efficacy following direct immunization. In this study, Mongolian gerbils were immunized with a bivalent inactivated EV71 C4a and inactivated CVA16 vaccine to test their effectiveness against viral infection. Bivalent vaccine immunization resulted in increased Ag-specific IgG antibody production; specifically, EV71 C4a-specific IgG was increased with medium and high doses and CVA16-specific IgG was increased with all doses of immunization. When gene expression of T cell-biased cytokines was analysed, Th1, Th2, and Th17 responses were found to be highly activated in the high-dose immunization group. Moreover, bivalent vaccine immunization mitigated paralytic signs and increased the survival rate following lethal viral challenges. When the viral RNA content was determined from various organs, all three doses of bivalent vaccine immunization were found to significantly decrease viral amplification. Upon histologic examination, EV71 C4a and CVA16 induced tissue damage to the heart and muscle. However, bivalent vaccine immunization alleviated this in a dose-dependent manner. These results suggest that the bivalent inactivated EV71 C4a/CVA16 vaccine could be a safe and effective candidate HFMD vaccine.

• 한국식품영양과학회지
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• 제46권5호
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• pp.537-544
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• 2017

본 연구에서는 참까막살 에탄올 추출물(PAEE)의 항염증 활성을 확인하기 위하여 LPS로 자극된 RAW 264.7 세포에서의 pro-inflammatory cytokine 및 NO의 분비 생성량과 western blot으로 단백질 발현량을 측정하였다. 또한, croton oil로 유도된 귀 부종 모델을 이용하여 알아보았다. RAW 264.7 세포에서 PAEE를 $0.1{\sim}100{\mu}g/mL$ 농도로 처리 시 세포독성을 나타내지 않음을 확인하였다. 그 결과 PAEE는 pro-inflammatory cytokine(IL-6, $TNF-{\alpha}$, $IL-1{\beta}$) 및 NO의 분비량을 농도 의존적으로 억제시켰으며, iNOS와 COX-2의 발현량도 감소시킴을 확인하였다. 이러한 항염증 활성결과는 $NF-{\kappa}B$와 MAPKs 전사인자의 활성 억제에 의한 것으로 확인되었다. 또한, croton oil로 유도된 귀 부종 모델에서 PAEE를 50 mg/kg body weight 처리 시 귀 부종이 prednisolone(10 mg/kg body weight)과 유사한 정도로 억제됨을 확인하였다. 귀 조직 관찰에서도 PAEE는 croton oil에 의해 증가한 진피와 경피의 두께를 감소시켰으며, 진피로 침윤된 mast cell의 수도 감소시켰다. 이 결과를 종합해 보면 참까막살 에탄올 추출물은 $NF-{\kappa}B$와 MAPKs의 활성화 억제를 통해 염증 매개 물질의 생성을 억제시켜 항염증 활성을 나타내는 것으로 확인되었다.

• Journal of Microbiology and Biotechnology
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• 제27권9호
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• pp.1628-1638
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• 2017

Viola tianshanica Maxim, belonging to the Violaceae plant family, is traditionally used in Uighur medicine for treating pneumonia, headache, and fever. There is, however, a lack of basic understanding of its pharmacological activities. This study was designed to observe the effects of the ethanol extract (TSM) from Viola tianshanica Maxim on the inflammation response in acute lung injury (ALI) induced by LPS and the possible underlying mechanisms. We found that TSM (200 and 500 mg/kg) significantly decreased inflammatory cytokine production and the number of inflammatory cells, including macrophages and neutrophils, in bronchoalveolar lavage fluid. TSM also markedly inhibited the lung wet-to-dry ratio and alleviated pathological changes in lung tissues. In vitro, after TSM ($12.5-100{\mu}g/ml$) treatment to RAW 264.7 cells for 1 h, LPS ($1{\mu}g/ml$) was added and the cells were further incubated for 24 h. TSM dose-dependently inhibited the levels of proinflammatory cytokines, such as NO, $PGE_2$, $TNF-{\alpha}$, IL-6, and $IL-1{\beta}$, and remarkably decreased the protein and mRNA expression of $TNF-{\alpha}$ and IL-6 in LPS-stimulated RAW 264.7 cells. TSM also suppressed protein expression of $p-I{\kappa}Ba$ and p-ERK1/2 and blocked nuclear translocation of $NF-{\kappa}B$ p65. The results indicate that TSM exerts anti-inflammatory effects related with inhibition on $NF-{\kappa}B$ and MAPK (p-ERK1/2) signaling pathways. In conclusion, our data demonstrate that TSM might be a potential agent for the treatment of ALI.

• 한국자원식물학회지
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• 제28권2호
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• pp.145-152
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• 2015

본 연구에서는 까마귀쪽나무 잎과 열매 추출물의 항염증 활성을 비교하고자 마우스 대식세포주인 RAW 264.7 cell에서 LPS에 의해 유도된 COX-2/PGE2와 iNOS/NO 및 염증성 cytokine 인 TNF-${\alpha}$, IL-$1{\beta}$, IL-6의 생성 억제 정도를 확인하였다. 그 결과, 동일한 농도 조건($10{\mu}g/ml$)에서 까마귀쪽나무의 열매 추출물이 잎 추출물에 비해 NO, $PGE_2$, iNOS, 염증성 cytokine인 TNF-${\alpha}$, IL-$1{\beta}$, IL-6의 생성량 모두에서 억제 효능이 우수하게 나타났다. 따라서 까마귀쪽나무가 보여주는 항염증 효과적 측면과 안전성의 두가지 측면을 고려할 때, 까마귀쪽나무의 열매 추출물이 저농도에서 잎 추출물보다 항염증 효과가 우수하거나 동등하며, 그 안전성이 크다고 판단됨으로 까마귀쪽나무의 열매추출물이 항염증 효능을 가진 기능성 식품 소재로써 개발 가능성이 있음을 제시한다.

• 동의생리병리학회지
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• 제26권4호
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• pp.511-518
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• 2012

The mechanisms of Toll-like receptor (TLR) signaling have been the focus of extensive studies because TLRs are the target of therapeutic intervention on multiple diseases. In this study, we investigated the inhibitory potential of Vigna angularis (azuki bean) on the TLR signaling. The effect of Vigna angularis extract (JSD) on TLR activation was investigated by assessing NF-${\kappa}B$ and AP-1 inducible secreted embryonic alkaline phosphatase (SEAP) activity. JSD significantly inhibited SEAP activity induced by poly I:C (TLR3 ligand) and poly I (TLR7 ligand) in a dose-dependent manner at concentration below 100 ${\mu}g/ml$ with no sign of cytotoxicity. Pretreatment of JSD markedly suppressed mRNA expressions of pro-inflammatory cytokines and adhesive molecules such as TNF-${\alpha}$, IL-6, RANTES, MCP-1 and ICAM-1 induced by TLR ligands. It also diminished the phosphorylation of $I{\kappa}B$ kinase and $I{\kappa}B$, and followed by $I{\kappa}B$-mediated nuclear translocation of p50, p65, and phosphorylation of p38, JNK, and IRF signaling pathway. In conclusion, our results suggest that Vigna angularis has inhibitory activity on TLR-3 and -7 signaling and it can be further developed as a remedy in curing TLR-related multiple diseases.

• 동의생리병리학회지
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• 제21권1호
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• pp.209-213
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• 2007

In the recent, increased concern has been focused on the pharmacology and clinical utility of herbal extracts and derivatives as a drug or adjunct to chemotherapy and immunotherapy. Salicis Radicis Cortex, A decoction has been mainly used for improvement of ozena and a diuretic effect in oriental medicine, but there was no study on the molecular mechanism of Salicis Radicis Cortex as an immunomodulator. Here we investigated the role of the aqueous extract of Salicis Radicis Cortex in the expression of inflammatory mediators, surface molecule, and related receptors in vitro and in vivo. In murine macrophage RAW 264.7 cells and peritoneal macrophages of C57BL/6N mice, water extract of Salicis Radicis Cortex increased the production of secretary TNF-alpha and Nitric oxide, and the expression level of CD14, LPS co-receptor and CD86, co-stimulatory molecule compared to negative natural extract ex vivo. Moreover, i.p. injection of water extract of Salicis Radicis Cortex significantly increased the secretion level of IFN-gamma and TNF-alpha, IL-2, IL-4 and IL-5 in serum of mice in vivo. Taken together, these results suggest that Salicis Radicis Cortex may regulate the immune response by secreting Th1 and Th2 types of cytokines in vivo and the possibility of its as natural immunostimulator.

• Journal of Microbiology and Biotechnology
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• 제10권2호
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• pp.147-153
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• 2000

This study was conducted to compare probiotic activities and physiological functions of Bifidobacterium longum Mk-G7 with weveral commercial and type strains of bifidobacteria. bif. longum MK-G7 showed the highest acid tolerance against HCl and acetic acid, whereas bif. infantis Y-1 showed the lowest acid tolerance and more than 4 log cycles of viable cell count decreased due to acid injuty. Viable cell counts of bifidobacteria strains decreased more than 1.5 log cycles owing to oxygen toxicity, with the exception of Bif. longum MK-G7, Bif. infantis Y-2, Bif. longum Y-3, Bif. longum Y-6, and Bif. longum RD-13 showed the highest bile tolerance, whereas Bif. longum MK-G7 showed a medium level of bile tolerance. Only Bif. longum MK-G7 howed much higher antibiotic resistance against both tetracycline and penicillin-G in the MIC(minimum inhibitory concentration) level of 24.8 mg/I and 0.52mg/I, respectively. Bif longum Y-6, and Bif. bifidum ATCC 29539 showed more than 80% of anti-mutagenicity against NQO(4-nitroquinolinel-oxide). Since the production of cytokines such as $TNF(tumor necrosis factor)-{\alpha}$ and IL (interleukin)-6, and NO(nitric oxide) in the macrophage cell line Raw 264.7 cells increased as Bif. longum MK-G7 cell concentration increased, ti was suggested that Bif. longum MK-G7 is able to enhance immunopotentiating activity in vitro. When freeze-dred Bif. longum MK-G7 was administered to mice at the dose of 1,2,4, and 6 g/kg of body weight, all of the mice survived in all feeding groups, proving the GRAS(generally recognized as safe) status of Bif. longum MK-G7. When fermented milk containing Bif. longum MK-G7 was administered to human volunteers, viable cell count of total bifidobacteria and anaerobes in the feces increased up to 0.5 log cycles more than before the administration. In particular, Bif. logum MK-G7 ingibited the growth of Bacteroides at the level of 1.0-1.5 log cycles.

• 대한의생명과학회지
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• 제12권3호
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• pp.289-301
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• 2006

Cardiopulmonary bypass (CPB) for cardiac surgery triggers the production and release of numerous chemotactic substances and cytokines, ensuing systemic inflammatory response that leads to postoperative major organ dysfunction. Traditionally, corticosteroids (steroid) have been administered to patients undergoing cardiac surgery to ward off these detrimental physiologic alterations. However, the majority of the studies have been performed on adult patients with high-dose steroid. We carried out a randomized, prospective, double-blind study to compare the efficacy of low-dose steroid with that of high-dose steroid and to determine the adequate dose of pretreated-steroid for prophylactic effects in pediatric cardiac surgery. Thirty pediatric patients scheduled for elective cardiac surgery were randomly assigned to two groups; fifteen patients received low-dose methylprednisolone (10mg/kg intravenously, n=15, low-dose group) and the others received high-dose methylprednisolone (30mg/kg intravenously, n=15, high-dose group) 1 hour prior to CPB. Arterial blood samples were taken before CPB (Pre-CPB), 10 minutes after start of CPB (CPB-10), and immediately after CPB-end (CPB-OFF) for measuring total leukocyte counts (T-WBC) and diff-counts, platelet counts, interleukin-6 (IL-6), myeloperoxidase (MPO), total antioxidant (TAO), neuron-specific enolase (NSE), troponin I (TNI), aspartate aminotransferase (AST), alanine aminotransferase (ALT), creatinine, and blood urea nitrogen (BUN) levels. Other parameters such as volumes of urine output, pulmonary index $(PI,\;PaO_2/FiO_2)$, mechanical ventilating period, intensive care unit (ICU)-staying period, postoperative complications (fever, wound problem), postoperative 24 hrs and total volumes in blood loss, and hospitalized days were also assessed. All parameters were compared between two groups. There were no significant differences in T-WBC counts, monocyte fraction, platelet counts, TA levels, NSE levels, creatinine levels, BUN levels, the volumes of total urine output, PI, the incidences of fever and wound problem, postoperative 24hrs- and total-blood loss volumes and ICU-staying period between two groups (P>0.05). At CPB-OFF, neutrophil fraction, MPO level, TNI level, and AST level were higher in the high-dose group than in the low-dose group (P<0.05). IL-6 level at CPB-10 was higher in the high dose-group than in the low-dose group (P<0.05). Furthermore, mechanical ventilating periods and hospitalized days of the high-dose group were significantly longer than those of low-dose group (P<0.05). The high-dose group had significantly low lymphocyte fi-action at CPB-OFF compared with the low-dose group (P<0.001). These findings suggest that pretreatment of high-dose steroid is not superior to that of low-dose steroid regrading its potential benefits in pediatric cardiac surgery. Therefore, the conventional strategy of steroid treatment, high-dose pretreatment, should be modified in the cardiac surgery with CPB. However, further studies must be performed on the larger number of patients in as much as small number of patients in this study.

• 한국미생물·생명공학회지
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• 제45권2호
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• pp.101-109
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• 2017

본 실험에서는 참치 심장 70% ethanol 추출물을 dichloromethane으로 분획한 후 항염증 효과를 확인하기 위해 RAW 264.7 세포에 LPS로 염증을 유도시켜 염증 매개성 물질인 NO와 pro-inflammatory cytokine의 분비량의 변화를 확인하였다. 그 결과, 참치 심장 dichloromethane 분획물을 처리하였을 때, 농도 의존적으로 NO의 생성량을 감소시키는 것을 확인하였으며, 특히, $100{\mu}g/ml$에서 가장 높은 억제효과를 나타내었다. 따라서 dichloromethane 분획물의 억제 활성이 세포사멸에 의한 감소인지 알아보기 위해서 MTT assay를 하였을 때, 세포 생존율이 dichloromethane 분획물을 PBS 처리군과 비교하였을 때 유의적인 차이가 나타나지 않음을 확인하였고, 이를 통해 dichloromethane 분획물이 NO 및 전염증성 cytokine의 분비를 효과적으로 억제할 수 있는 물질임을 확인할 수 있었다. Dichloromethane 분획물을 처리하였을 때, 염증 관련 단백질 발현 정도를 western blot을 통해 확인한 결과, LPS에 의해 발현이 증가된 $NF-{\kappa}B$, iNOS 및 COX-2는 분획물을 처리함으로써 농도 의존적으로 감소되는 것을 확인할 수 있었다. 또한 dichloromethane 분획물의 처리가 인산화된 MAPKs의 발현을 저해함을 확인하여 참치 심장 dichloromethane 분획물이 $NF-{\kappa}B$와 MAPKs의 발현을 억제시킴으로써 NO 및 pro-inflammatory cytokine의 분비량을 감소시킴을 확인할 수 있었다. 동물 모델에서는, dichloromethane 분획물을 처리하였을 때 croton oil에 의한 귀 부종이 농도 의존적으로 감소함을 확인하였고, 특히, $250mg/kg{\cdot}body\;weight$ 농도로 투여시 시판 항염증제인 predinisolone을 $50mg/kg{\cdot}body\;weight$ 농도로 투여한 그룹과 유사한 효과를 나타내었다. 조직학적 변화를 확인한 결과에서는, 진피와 경피의 두께가 감소하였으며 진피내 mast cell 침윤이 감소되는 것을 확인할 수 있었다. 따라서 참치심장 dichloromethane 분획물이 효과적인 염증 예방 및 부종 완화를 위한 치료제로서 활용 가능성을 확인하였다.