SCI 발표논문

SCI 발표논문

← SCI 발표논문 목록

PLoS One. 2014;9(9):e106861.

Chronic low dose chlorine exposure aggravates allergic inflammation and airway hyperresponsiveness and activates inflammasome pathway.

, , , ,

제1저자 김세훈 · 교신저자 강혜련, 조상헌

PMID 25202911 PMC4159271 DOI 10.1371/journal.pone.0106861 IF 2.8 알레르기내과
소속 (3)
  1. Department of Internal Medicine, Seoul National University College of Medicine, Seoul, Republic of Korea; Institute of Allergy and Clinical Immunology, Seoul National University Medical Research Center, Seoul, Korea; Department of Internal Medicine, Seoul National University Bundang Hospital, Seongnam, Republic of Korea.
  2. Institute of Allergy and Clinical Immunology, Seoul National University Medical Research Center, Seoul, Korea.
  3. Department of Internal Medicine, Seoul National University College of Medicine, Seoul, Republic of Korea; Institute of Allergy and Clinical Immunology, Seoul National University Medical Research Center, Seoul, Korea.

Abstract

BACKGROUND Epidemiologic clinical studies suggested that chronic exposure to chlorine products is associated with development of asthma and aggravation of asthmatic symptoms. However, its underlying mechanism was not clearly understood. Studies were undertaken to define the effects and mechanisms of chronic low-dose chlorine exposure in the pathogenesis of airway inflammation and airway hyperresponsiveness (AHR).

METHODS Six week-old female BALB/c mice were sensitized and challenged with OVA in the presence and absence of chronic low dose chlorine exposure of naturally vaporized gas of 5% sodium hypochlorite solution. Airway inflammation and AHR were evaluated by bronchoalveolar lavage (BAL) cell recovery and non-invasive phlethysmography, respectively. Real-time qPCR, Western blot assay, and ELISA were used to evaluate the mRNA and protein expressions of cytokines and other inflammatory mediators. Human A549 and murine epithelial (A549 and MLE12) and macrophage (AMJ2-C11) cells were used to define the responses to low dose chlorine exposure in vitro.

RESULTS Chronic low dose chlorine exposure significantly augmented airway inflammation and AHR in OVA-sensitized and challenged mice. The expression of Th2 cytokines IL-4 and IL-5 and proinflammatory cytokine IL-1β and IL-33 were significantly increased in OVA/Cl group compared with OVA group. The chlorine exposure also activates the major molecules associated with inflammasome pathway in the macrophages with increased expression of epithelial alarmins IL-33 and TSLP in vitro.

CONCLUSION Chronic low dose exposure of chlorine aggravates allergic Th2 inflammation and AHR potentially through activation of inflammasome danger signaling pathways.

Figures

Figure 1
Figure 1 Mouse chlorine exposure system. A, a container filled with NaOCl solution was placed in the cage and mice were exposed to naturally vaporizing chlorine gas 8 hours a day, 5 times a week, for 4 weeks. B, a container with holes on the top surface to avoid direct skin contact with the NaOCl solution.
Figure 2
Figure 2 Effect of low dose chronic chlorine exposure on allergen-induced airway hyperresponsiveness and inflammation 6 weeks old BALB/c mice were sensitized and challenged with OVA allergen with or without low dose chlorine exposure. A, Airway hyperresponsiveness was determined by whole body phlethysmography *p<0.05 vs. other groups. B, Lung inflammatory response were measured by BAL cell recovery. Macro, macrophages; Neutro, neutrophils; Eosino, eosinophils; Lympho, lymphocytes. *p<0.05 vs. other groups. C, H&E staining (x200). D, Quantification of eosinophil in the lung tissue. E, PAS staining (x200), F: Semi-quantified mucus index score in the PAS staining. Values in panels A, B, D and F are mean ± SEM of evaluations in a minimum of 5 mice. Panel C and E is a representative of a minimum of four similar experiments.
Figure 3
Figure 3 Effect of low dose chronic chlorine exposure on Th2 cytokine and IL-17 expression in OVA sensitized and challenged mice. A-C, the mRNA expression of IL-4 (A), IL-5(B) and IL-13(C) was evaluated by RT-qPCR. D. The level of IL-17 in BAL was measured by ELISA. Values in these panels are mean ± SEM of evaluations in a minimum of 5 mice. *p<0.05 vs. OVA only group.
Figure 4
Figure 4 Effect of low dose chronic chlorine exposure on the molecules associated with inflammasome activation. A, IL-1β levels in the BAL from OVA-sensitized and challenged mice with or without chlorine exposure were measured by ELISA. *p<0.05 vs. OVA only group. B, The mRNA expressions of IL-1β were measured by RT-qPCR after 48 and 96 hrs stimulation with 0.001% NaOCl. *p<0.05 vs. control. C, IL-1β levels were measured using ELISA after low dose NaOCl treatments in AMJ2-C11 mouse macrophage cells. *p<0.05 vs. control. D, Western blot evaluation on the molecules associated with inflammasome activation in the macrophages with and without exposure of NaOCl (IL-1β and caspase-1 in the cell supernatant, and IL-18 in the cell lysates). Values in panels in A, B and C are mean ± SEM. Panel D is a representative of a minimum of three similar experiments.
Figure 5
Figure 5 Effect of low dose chronic chlorine exposure on the expression of epithelial alarmines. A, The levels of IL-33 expression in the lungs from the mice sensitized and challenged by OVA with or without chlorine exposure were evaluated by RT-qPCR. *p<0.05 vs. groups without chlorine exposure. B, The mRNA expressions of IL-33 in murine MLE12 epithelial cells were measured by qRT-PCR after 48 and 96 hrs stimulation of 0.001% NaOCl. *p<0.05 vs. control. C, TSLP protein expression was evaluated by Western blot analysis. Values in panels A and B are mean ± SEM of evaluations in a minimum of 5 mice. Panel C is a representative of a minimum of three similar experiments.

그림 출처: PubMed Central 공개 논문 (PMC4159271)

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
2014-09-09
MeSH
Animals · Asthma / chemically induced / genetics / immunology / pathology · Chlorine / adverse effects · Cytokines / genetics · Dose-Response Relationship, Drug · Epithelial Cells / drug effects / metabolism · Female · Gene Expression Regulation / drug effects · Humans · Inflammasomes / metabolism · Interleukin-33 · Interleukins / genetics · Mice · Mice, Inbred BALB C · Respiratory Hypersensitivity / chemically induced / genetics / immunology / pathology · Risk Factors · Signal Transduction / drug effects · Th2 Cells / drug effects / metabolism · Time Factors · Thymic Stromal Lymphopoietin
Substances
Cytokines · Il33 protein, mouse · Inflammasomes · Interleukin-33 · Interleukins · Chlorine · Thymic Stromal Lymphopoietin
참고문헌
45편