SCI 발표논문

SCI 발표논문

← SCI 발표논문 목록

PLoS One. 2015;10(4):e0123094.

Thalidomide inhibits alternative activation of macrophages in vivo and in vitro: a potential mechanism of anti-asthmatic effect of thalidomide.

, , , , , , , , ,

제1저자 이현승, 권혁수 · 교신저자 강혜련, 장윤석

PMID 25905462 PMC4408009 DOI 10.1371/journal.pone.0123094 IF 2.8 알레르기내과
소속 (5)
  1. Institute of Allergy and Clinical Immunology, Seoul National University Medical Research Center, Seoul, Korea.
  2. Institute of Allergy and Clinical Immunology, Seoul National University Medical Research Center, Seoul, Korea; Department of Internal medicine, Seoul National University College of Medicine, Seoul, Korea.
  3. Department of Medical Science, Seoul National University College of Medicine, Seoul, Korea.
  4. Department of Anatomy, Seoul National University College of Medicine, Seoul, Korea.
  5. Department of Internal Medicine, Seoul National University Bundang Hospital, Seongnam, Gyeonggi-do, Korea.

Abstract

BACKGROUND Thalidomide is known to have anti-inflammatory and immunomodulatory actions. However, the effect and the anti-asthmatic mechanism of thalidomide in the pathogenesis of asthmatic airways are not fully understood.

OBJECTIVE This study is designed to determine the effect and the potential mechanism of thalidomide in the pathogenesis of asthmatic airways using animal model of allergic asthma.

METHODS Six-week-old female BALB/C mice were sensitized with alum plus ovalbumin (OVA) and were exposed to OVA via intranasal route for 3 days for challenge. Thalidomide 200 mg/kg was given via gavage twice a day from a day before the challenge and airway hyperresponsivenss (AHR), airway inflammatory cells, and cytokines in bronchoalveolar lavage fluids (BALF) were evaluated. The expression levels of pro-inflammatory cytokines and other mediators were evaluated using ELISA, real time (RT)-qPCR, and flow cytometry. CRL-2456, alveolar macrophage cell line, was used to test the direct effect of thalidomide on the activation of macrophages in vitro.

RESULTS The mice with thalidomide treatment showed significantly reduced levels of allergen-induced BALF and lung inflammation, AHR, and the expression of a number of pro-inflammatory cytokines and mediators including Th2 related, IL-17 cytokines, and altered levels of allergen-specific IgG1/IgG2a. Of interesting note, thalidomide treatment significantly reduced expression levels of allergen- or Th2 cytokine-stimulated alternative activation of macrophages in vivo and in vitro.

CONCLUSION These studies highlight a potential use of thalidomide in the treatment of allergic diseases including asthma. This study further identified a novel inhibitory effect of thalidomide on alternative activation of macrophages as a potential mechanism of anti-asthmatic effect of thalidomide.

Figures

Fig 1
Fig 1 Effects of thalidomide on lung inflammation and AHR by allergen challenges. A, Experimental protocol of the study. B, The number of inflammatory cells including eosinophils in BALF. C, H&E stain (a-d), ×100) and PAS stain (e-h), ×200) of lung histology after allergen challenge (a,e: PBS; b,f: PBS+Thalidomide; c,g: OVA; d,h: OVA+Thalidomide). * indicates p < 0.05. (D) Methacholine hyperresponsiveness was measured 24 h after the last intranasal challenge, n = 5–8 for each group, the ** indicates p < 0.01. Mch; methacholine bronchial challenge.
Fig 2
Fig 2 Effects of thalidomide on serum allergen-specific antibodies. A-C, The levels of serum OVA-specific IgG2a and IgG1 48h after the last challenge. Serum OVA-specific IgG1 (A), IgG2a levels (B) and IgG1/IgG2a ratio (C) after OVA challenge. n = 5–8 for each group, * indicates p < 0.05.
Fig 3
Fig 3 Effects of thalidomide on cytokine protein levels in BALF, Lung draining lymph nodes and spleen. A-C, The upper low shows the levels of IL-4(A), IL-5(B) and IL-17(C) 48h after the last challenge. The levels were detected with enzyme-linked immunosorbent assay. D-G, The lower low shows the frequencies of IL-4(D) and IL-5(E)-producing CD4+ T cells 72h after re-stimulated in vitro with OVA protein in draining lymph node cells. The frequencies of IL-4(F) and IFN-γ(G)-producing CD4+ T cells 72h after re-stimulated in vitro with OVA protein in spleen cells. The expressions were analyzed using Flow cytometry. n = 5–8 for each group, * indicates p < 0.05.
Fig 4
Fig 4 Effects of thalidomide on the mRNA expressions in mouse lung tissues. A-D, Real-time RT-PCR was performed to determine the changes in mRNA levels of TNF-α (A), IL-13(B), Eotaxin-1(C) and MUC5AC(D) from lung tissues. The levels of mRNA are represented as the ratio to β-actin. n = 5–8 for each group, * indicates p < 0.05.
Fig 5
Fig 5 Effects of thalidomide on the mRNA expressions of M2 macrophage markers in mouse lung tissues. A-D, Real-time RT-PCR was performed to determine the changes in mRNA levels of Relm-α(A), Arg-1(B), CD206(C) and Ym-1(D) from lung tissues. The levels of mRNA are represented as the ratio to β-actin. n = 5–8 for each group. * indicates p < 0.05, ** indicates p < 0.01.
Fig 6
Fig 6 Effects of thalidomide on the populations of M2 macrophage markers in mouse lung tissues. A-C, Representative dot plot shows the expression of F4/80(A) in CD45+ cells from lung and CD206(B) and Ym-1(C) in F4/80+ macrophage cells. D-F, The frequencies of total F4/80+ macrophage cells(D) in CD45+ cells, CD206(E), and Ym-1(F) producing F4/80+ macrophage cells are shown. G-H, The levels of M1 (F4/80+ CD11c+CD206-) cells are represented as the frequencies(G) and the CD206 /M1 cells ratio(H) are represented. n = 5–8 for each group. * indicates p < 0.05
Fig 7
Fig 7 Effects of thalidomide on the expressions of M2 macrophage markers in mouse alveolar macrophages cell line. A-C, The mRNA levels of Relm-α(A), Arg-1(B), and CD206(C) were detected in Th2 cytokines stimulated mouse alveolar macrophages cell line. The levels of mRNA are represented as the ratio to β-actin. D-E, The frequencies of CD206(D) or Ym-1(E) were detected in Th2 cytokines stimulated mouse alveolar macrophages cell line using Flow cytometry. * indicates p < 0.05, ** indicates p < 0.01.

그림 출처: PubMed Central 공개 논문 (PMC4408009)

Conflict of interest statement

Competing Interests:The authors have declared that no competing interests exist.

그 밖의 정보
출판 형태
Journal Article, Research Support, Non-U.S. Gov't
저널
PloS one · eISSN 1932-6203
전자 출판
2015-04-23
MeSH
Allergens / pharmacology · Alum Compounds / pharmacology · Animals · Anti-Asthmatic Agents / pharmacology · Anti-Inflammatory Agents / pharmacology · Asthma / chemically induced / drug therapy / metabolism · Bronchial Hyperreactivity / chemically induced / drug therapy / metabolism · Bronchoalveolar Lavage Fluid · Disease Models, Animal · Female · Immunoglobulin G / metabolism · Inflammation / chemically induced / drug therapy / metabolism · Interleukin-17 / metabolism · Macrophages, Alveolar / drug effects / metabolism · Mice · Mice, Inbred BALB C · Ovalbumin / pharmacology · Th2 Cells / drug effects / metabolism · Thalidomide / pharmacology
Substances
Allergens · Alum Compounds · Anti-Asthmatic Agents · Anti-Inflammatory Agents · IL17A protein, human · Immunoglobulin G · Interleukin-17 · ovalbumin-alum · Thalidomide · Ovalbumin
참고문헌
42편