• 대한한의학회지
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• 제30권1호
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• pp.150-162
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• 2009

Objectives: Inflammatory mediators, such as nitric oxide (NO), prostaglandin E2 ($PGE_2$) and pro-inflammatory cytokines, TNF-${\alpha}$ and IL-$1{\beta}$ playa critical role in inflammatory immune response. Therefore, intervention of inflammatory mediator production promises therapeutic benefit for treatment of many chronic inflammatory diseases, such as allergic asthma, rheumatoid arthritis, multiple sclerosis, septic shock and neurodegenerative diseases. In this study, the pharmacological effects of the flower MeOH extract Panax notoginseng (Notoginseng Flos; NF) on inflammation were investigated to address potential therapeutic or toxic effects. Methods: RA W264.7 cells were treated with different concentrations of NF methanol (NF-M) extract in the presence or absence of LPS ($1{\mu}g/m{\ell}$). Results: NF-M extract significantly inhibited LPS-induced production of NO, $PGE_2$ and pro-inflammatory cytokines, TNF-${\alpha}$ and IL-$1{\beta}$ in a dose-dependent manner. In addition, NF-M extract suppressed mRNA expressions and protein levels of iNOS, COX-2 and pro-inflammatory cytokines in LPS-stimulated RA W264.7 cells. Conclusion: These results indicated that NF-M extract inhibits LPS-induced production of inflammatory mediators in macrophages and demonstrated that NF-M extract possesses anti-inflammatory properties in vitro.

• IMMUNE NETWORK
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• 제21권6호
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• pp.42.1-42.20
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• 2021

Eosinophils play critical roles in the maintenance of homeostasis in innate and adaptive immunity. Although primarily known for their roles in parasitic infections and the development of Th2 cell responses, eosinophils also play complex roles in other immune responses ranging from anti-inflammation to defense against viral and bacterial infections. However, the contributions of pattern recognition receptors in general, and NOD-like receptors (NLRs) in particular, to eosinophil involvement in these immune responses remain relatively underappreciated. Our in vivo studies demonstrated that NLRC4 deficient mice had a decreased number of eosinophils and impaired Th2 responses after induction of an allergic airway disease model. Our in vitro data, utilizing human eosinophilic EoL-1 cells, suggested that TLR2 induction markedly induced pro-inflammatory responses and inflammasome forming NLRC4 and NLRP3. Moreover, activation by their specific ligands resulted in caspase-1 cleavage and mature IL-1β secretion. Interestingly, Th2 responses such as secretion of IL-5 and IL-13 decreased after transfection of EoL-1 cells with short interfering RNAs targeting human NLRC4. Specific induction of NLRC4 with PAM3CSK4 and flagellin upregulated the expression of IL-5 receptor and expression of Fc epsilon receptors (FcεR1α, FcεR2). Strikingly, activation of the NLRC4 inflammasome also promoted expression of the costimulatory receptor CD80 as well as expression of immunoregulatory receptors PD-L1 and Siglec-8. Concomitant with NLRC4 upregulation, we found an increase in expression and activation of matrix metalloproteinase (MMP)-9, but not MMP-2. Collectively, our results present new potential roles of NLRC4 in mediating a variety of eosinopilic functions.

• IMMUNE NETWORK
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• 제1권2호
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• pp.116-125
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• 2001

Background: Nitric oxide (NO) production has been described as a double-edged sword eliciting both pro- and anti-inflammatory effects in different immune reactions. This work was undertaken to investigate the immunoregulatory role of NO in experimental allergic encephalomyelitis (EAE) and experimental allergic uveitis (EAU). Method: We examined whether molsidomine (MSDM), a NO donor, administration to the myelin basic protein (MBP)- or interphotoreceptor retinoid binding protein (IRBP)-immunized rats could suppress EAE development by shifting toward the Th2 cytokine response. In the EAE experiments, the rats were treated orally with MSDM (10 mg/kg/day) at the early stage (-1~4 days) or throughout the experimental period (-1~15 days). Results: This resulted in significant amelioration of the disease and mild clinical symptoms, while MBP-immunization without MSDM administration showed severe EAE development. A marked reduction in inflammation was also observed in the spinal cord, indicating the crucial role of NO in the pathogenesis of EAE in in vivo. In the EAU experiments, a 24 h pre-treatment with MSDM prior to IRBP immunization resulted in significant inhibition of the disease. Furthermore, MSDM administration for 2 1 days completely reduced the incidence and severity of EAU. To investigate whether MSDM could modulate cytokine switching from Th 1 to Th2, culture supernatants of MBP- or IRBP-stimulated inguinal lymphocytes were analyzed. MSDM treatment enhanced IL-10 secretion but decreased IFN-${\gamma}$. IL-4 was undetectable in all groups. In contrast, the MBP-or IRBP-immunized rats without MSDM secreted high concentrations of IFN-${\gamma}$, but low concentrations of IL-10. Conclusion: In conclusion, NO administation suppresses EAE and EAU by modulating the Th1/Th2 balance during inflammatory immune responses. This work further suggests that NO may be useful in the therapeutic control of autoimmune disease.

• 대한한방내과학회지
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• 제31권2호
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• pp.189-200
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• 2010

Objectives : Regulatory T cells can reduce inflammation and allergic reactions through their inhibitory functions. Gardeniae Fructus(GF) is a Heat-clearing herb used in traditional Korean medicine, and a wide range of studies on its antiinflammatory effects are being carried out. The authors investigated the effect that Gardeniae Fructus has on regulatory T cells. Methods : The authors screened 14 herbs for their effects on regulatory T cells. 100mg of each herb were separately dissolved in 1ml of sterile saline and the supernatant was harvested after 10 minutes of centrifuge at 15,000 rpm. The supernatant was filtered through a 0.2 ${\mu}m$ syringe filter, and the resulting stock was refrigerated at $4^{\circ}C$. The stock was diluted before testing and used at a final concentration of $0.01{\mu}g/ml$. CD4+CD25+ T cells from healthy BALB/c spleens were used as natural regulatory T cells (nTreg), and CD4+CD25- T cells were used as reactive T cells. CD4+CD25+ and CD4+CD25- T cells were activated with anti-CD3e ($10{\mu}g/m{\ell}$)/anti-CD28 ($1{\mu}g/m{\ell}$) and cultured. IL-10 from supernatant of the culture medium was measured by IL-10 cytokine ELISA. The percentages, cell numbers, phenotype and function of CD4+CD25+ Treg cells were determined by flow cytometry. Results : Gardeniae Fructus was shown to be the most potent herb among the 14 herbs tested for suppressing CD4+CD25- reactive T cell proliferation by stimulating CD4+CD25+ natural regulatory T cells. Gardeniae Fructus induces IL-10 secretion increase by stimulating CD4+CD25+ natural regulatory T cells, and indirectly suppresses CD4+CD25- reactive T cell proliferation through increasing CD25 (IL-2 receptor $\alpha$) expression and thus promoting bonding with IL-2. Gardeniae Fructus did not directly affect CD4+CD25- reactive T cell proliferation. Conclusions : Gardeniae Fructus suppressed reactive T cell proliferation through inducing increases in IL-10 secretion and CD25 (IL-2 receptor $\alpha$) expression.

• 혜화의학회지
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• 제18권1호
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• pp.29-41
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• 2009

Wished to examine closely effect that SoPungDoJeokTang-KaMi (SPDJTK) medicines used to atopy dermatitis disease patient get in atopy eruption control experimentally. SPDJTK medicines controlled $CD4^+/IFN-\gamma$, and $CD4^+/CD25^+/foxp3^+$ revelation that an experiment that motive allergy immune reponse because an in vitro experiment stimulates T cells of a NC/Nga mouse same time by anti-CD40/rmIL-4, and interleukin-$1{\beta}$, IL-6, TNF-$\alpha$, and TGF-$\beta$ mRNA outturn that bear in T and B cells decreased remarkably by SPDJTK medicines. Intracellular staining of splenocytes anti-CD40/rmIL-4 plus rmIL-4 stimulated as described in a, assessed after 24 h, SPDJTK exerts a mainly immunosuppressive effect that acts at least partially through suppression of the transcription factor GATA3 expression in $CD4^+$ T cells. Atopic dermatitis (AD) usually develops in patients with an individual or family history of allergic diseases, and is characterized by chronic relapsing inflammation seen specially in childhood, association with IgE hyperproduction and precipitation by environmental factors. However, the exact etiology of AD has been unclear. To further explore the pathogenesis and treatment of AD, a suitable animal model is required. We found that skin lesions, which were clinically and histologically very similar to human AD, mite antigen-induced dermatitis on the face, neck, ears and dorsal skin of inbred NC/Nga mice. Result that Th1 cell and Th2 cell observe to be shifted by cytokine expression with $CD4^+/CD25^+/foxp3^+$ Treg cells induction by SPDJTK medicines could know that SPDJTK medicines can use usefully in allergy autoimmnune diease.

• 생명과학회지
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• 제23권9호
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• pp.1109-1117
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• 2013

본 연구에서는 인체 폐암 세포인 A549를 사용하여 택란 메탄올 추출물의 항암활성과 그 분자적 기전에 관하여 연구하였다. 먼저 택란 추출물이 A549의 세포증식에 미치는 영향을 알아본 결과 처리 농도 및 시간 의존적으로 A549의 성장이 저해되었으며, 세포 주기 변화를 분석한 결과 강력한 G1 arrest가 유도되는 것을 확인하였다. 이러한 택란 추출물에 의한 G1 arrest는 세포주기 조절 단백질인 Cyclin D1, Cyclin E 및 Cyclin-dependent kinase인 CDK2, CDK4, CDK6의 발현 감소와 연관되어 있었다. 또한 택란 추출물에 의한 CDK/Cyclin complex의 발현 저해는 DNA 손상에 의해 활성화되는 CHK2의 활성화 형태인 p-CHK2의 발현 증가에 따른 CDK 활성화 효소인 Cdc25A phosphatase의 발현 억제에 의해 나타나는 결과로 사료된다. 반면 종양억제유전자인 p53 및 CDK 억제제인 p21과 p27의 발현량은 증가되지 않았다. 이러한 결과들로부터 택란 추출물은 DNA damage에 의한 ATM/CHK2/Cdc25A/CDK2 pathway를 통해 A549의 G1 arrest를 유도하여 세포의 증식을 억제할 것으로 판단되며, 이때 택란 추출물에 의해 유도되는 G1 arrest는 p53 비의존적인 경로일 것으로 사료된다. 본 연구결과는 택란이 Cdc25A를 target으로 하는 새로운 항암활성 소재로서 사용될 수 있는 가능성을 시사한다. 또한 본 연구결과는 택란 추출물의 세포주기 조절에 의한 항암기전을 이해하고 향후 지속적 연구를 하는 데 있어서 귀중한 기초자료로 사용될 수 있을 것이다.

• 대한화장품학회지
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• 제46권3호
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• pp.283-294
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• 2020

본 연구에서는 유자 완숙과의 중량비 13%를 차지하지만 대부분 폐기 처분되어 활용이 되지 않는 유자씨앗의 활용도를 높이기 위하여, 냉압착 방법으로 추출한 유자씨 오일의 피부미용 효능을 연구하였다. 유자씨 오일은 주로 oleic acid와 linoleic acid로 이루어진 불포화 지방산을 약 74% 함유 하는 것으로 확인되었으며, 주로 유자 과피에 포함되는 limonene은 약 0.0187%로 매우 낮은 함량을 포함하였다. 유자씨 오일의 DPPH radical 소거 활성능력을 평가한 결과, 5% 농도에서 26%의 DPPH 라디칼 소거능력을 확인하였다. 항염 효능을 확인하기 위해 유자씨오일에 대한 RAW 264.7 세포 독성 시험 및 NO 생성을 시험한 결과, 세포 독성을 나타내지 않는 0.05% 농도에서 NO 생성을 53% 억제하였다. 그리고 항알러지 효능확인을 위한 RBL-2H3 세포 독성 및 β-hexosaminidase 방출 억제 효능 시험에서도 세포 독성을 나타내지 않는 0.05% 농도에서 β-hexosaminidas 방출을 26% 억제함을 확인하였다. 마지막으로 유자씨 오일을 5% 함유한 O/W 에멀젼의 인체 적용 시험 결과에서는 caprylic/capric triglyceride를 동량 함유하는 대조 에멀젼에 비하여 높은 피부 보습 효능을 나타내었다. 따라서 유자씨 오일은 우수한 스킨케어 소재로 활용 될 수 있을 것으로 판단된다.

• 한방안이비인후피부과학회지
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• 제22권2호
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• pp.1-18
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• 2009

Objective : Moutan Cortex (the root bark of Paeonia suffruticosa Andr.) is widely used in oriental medicine as a remedy for inflammation. However, as yet there is no clear explanation of how MC(Moutan Cortex) affects the production of inflammatory cytokine. This study was to determine the effects of Essence extracted MC on the mast cell-mediated inflammatory responses. Method : We observed the effect of MC on compound 48/80-induced histamine release of rat peritoneal mast cells and the effect of administering MC on PCA in rat. We measured the amount of inflammatory cytokine production induced by the phorbol myristate acetate (PMA) plus calcium ionophore(A23187) in the human mast cell line (HMC-1) incubated with various concentrations of MC. The TNF-$\alpha$ protein levels were analysised by Western blot. The TNF-$\alpha$, IL-6 and IL-8 secreted protein levels were measured by the ELISA assay. The TNF-$\alpha$, IL-6 and IL-8 mRNA levels were measured by the RT-PCR analysis. NF-$\kappa$B, phospho-I$\kappa$B and MAPKs were exmined by Western blot analysis. The NF-$\kappa$B promoter activity was examined by luciferase assay. Result : 1. Enzyme immunoassay indicated that MC suppressed histamine secretion of rat peritoneal mast cells. 2. In PCA dependent on IgE, MC had anti-allergic effect of the internal surface of rat skin. 3. Western blot indicated that MC decreased TNF-$\alpha$ protein levels. 4. ELISA indicated that MC decreased TNF-$\alpha$, IL-6 but MC had no significant effect on IL-8 in HMC-1 cells. 5. RT-PCR indicated that MC decreased TNF-$\alpha$, IL-8 but MC had no significant effect on IL-6 in HMC-l cells. 6. Western blot indicated that MC suppressed the induction of MAPKs, NF-$\kappa$B & phospho-I$\kappa$B activity in HMC-1 cells. 7. Luciferase assay indicated that MC suppressed the PMA plus A23187-induced NF-$\kappa$B promoting activityin HMC-1 cells. Conclusion : In this study, we have found that MC is an inhibitor of NF-$\kappa$B, MAPKs & cytokines on the mast cell-mediated inflammatory responses.

• Tuberculosis and Respiratory Diseases
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• 제45권5호
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• pp.1012-1021
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• 1998

배 경: 기관지 천식은 기도에 발현된 복합적인 세포성 혹은 화학-매개물 등의 영향으로 기도내 만성적인 면역학적 염증을 보이는 기저병리를 특정으로 하며 이러한 결과로서 기도과민성을 포함한 기관지수축에 따른 호흡기 증세를 나타내므로 기도내 염증조절이 중요하게 인식되고 있다. 기침형 천식은 뚜렷한 호흡곤란이나 천명음없이 주로 만성기침만을 호소하며 기도 과민을 특징적으로 보인다. 스테로이드는 대표적인 항염증 흡입제로 인정되고 있으며 nedocromil sodium은 cromolyn에 비해 효과적인 항염증제로 알려져있다. 저자들은 기침형 천식환자들에서 전향적으로 흡입 항염증제 치료에 따른 기도과민도 변화 경과를 관찰하고 기도과민도 호전여부에 영향을 미치는 인자들을 평가하고자 하였으며 기침형 천식환자에서 nedocromil의 치료 효과를 평가하고자 하였다. 대상 및 방법: 기침형 천식환자 61명을 대상으로 전향적으로 조사하였다. 23명에서 budesonide ($400{\mu}g{\times}2/day$), 22명에서 nedocromil($4mg{\times}2/day$) 군에 속하였으며 16명에서 병합치료를 하였다. 기도과민성은 DeVilbiss 646 Dosimeter를 이용한 메타콜린 기관지 유발검사로 평가하였다. 증세변화여부는 본조사에서 정해진 양식에 따라 문진하여 증세-점수도를 산출하여 평가하였다. 각군에서 2 개월간 흡입치료후 시행된 기관지유발검사 결과 양성반응을 보이는 경우 임의로 비-관해군(non-responder)으로 정하여 이들을 대상으로 각각 치료후 4 개월 혹은 6개월째에 기관지유발검사를 재시행, 평가하였다. 결 과: 각군간의 평균 연령 및 성별에는 차이가 없었으며 기침발현 평균기간은 각각 7.5, 4.5, 5.4년으로 차이가 없었다. 각군에서 내원당시 평균흡연력 및 평균 FEV1% 치는 차이가 없었으며 내원당시 메타콜린 PC20(mg/ml) 평균치는 각각 3.0, 4.8, 2.9으로 각 군간에 차이는 없었다. 각군에서 내원당시 증세-점수도간에 차이는 없었다. 치료 2개월째 기도 과민도 관해를 보인 정도는 budesonide 군에서 30.4%, nedocromil 군 31.8%, 병합치료군에서 37.5%로 각 군간에 차이가 없었다(p=0.95). 각군에서 기도 과민도는 2개월 치료후 내원당시에 비해 각각 4.24, 2.44, 6.91 배로 유의하게 감소하였으며(p<0.05), 증세-점수도는 각군에서 유의하게 감소하여(p<0.05) 증세 호전을 보였다. 치료 2개월째 기도 과민도의 관해를 보이지 않았던 환자들에서 메타콜린 축적용량단위 및 증세-점수도는 내원당시에 비해 2개월째에 유의한 호전을 보였으나(p<0.01) 각각 4개월 혹은 6개월째 재평가한 결과 2개월째와는 차이가 없었다. 치료 2 개월째 기도 과민도 관해를 보인군에서 혈중 총 IgE, 말초혈액 호산구수 및 피부반응 양성율은 관해를 보이지 않았던 군과 차이가 없었으나 치료시작전 증세발현기간 정도, 흡연력, 연령, 성별 등에서는 양군간에 유의한 차이를 보였으며(p<0.05) 내원당시 메탈콜린 PC20 치는 관해군에서 유의하게 높은 결과를 보였다(p<0.01). 결 론: 치료 2개월째에 기도 과민도 관해를 보이지 못한 군에서는 이후의 관찰기간동안 기도 과민도 및 증세-점수도는 각각 2개월째와 차이가 없었으며 치료시작전 증세발현기간, 연령, 흡연력, 성별, PC20 등은 기도 과민도 관해에 유의한 영향을 미치는 것으로 사료되었다. 기침형 천식환자에서 병합치료가 budesonide나 nedocromil 단독치료에 비해 우월하지 않는 것으로 사료되었다.