Copper oxide nanoparticles (CuONPs), widely used across various industries, pose inhalation risks to industrial workers and consumers through respiratory exposure. Although exposure to CuONPs has been implicated in the initiation and progression of respiratory disorders, the molecular mechanisms underlying these effects remain poorly defined. In this study, we investigated the role of the nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway in CuONP-induced respiratory toxicity and asthma exacerbation. Exposure to CuONPs resulted in pronounced inflammatory cell infiltration, elevated cytokine production, and excessive mucus secretion, accompanied by disrupted redox balance, as evidenced by increased malondialdehyde (MDA) levels and decreased glutathione (GSH) concentrations. These changes were associated with upregulation of Nrf2 and its downstream antioxidant enzymes, including heme oxygenase-1 (HO-1) and glutamate-cysteine ligase modifier subunit (GCLM). Although this antioxidant response is consistent with an expected oxidative stress-adaptive pathway, it was not sufficient to prevent CuONP-associated inflammatory and redox disturbances. Consistently, in NCI-H292 cells, CuONP treatment increased the expression of Nrf2, HO-1, and GCLM, whereas siRNA-mediated Nrf2 knockdown abrogated these inductions. In an ovalbumin (OVA)-induced asthma model, CuONP exposure further intensified airway inflammation and oxidative stress, despite elevated Nrf2 expression. However, adeno-associated virus (AAV)-mediated Nrf2 overexpression significantly attenuated CuONP-induced airway inflammatory responses and redox imbalance in asthmatic mice. Taken together, our results indicate that endogenous Nrf2 response is insufficient to counteract CuONP-driven asthma exacerbation, whereas pharmacological or genetic augmentation of Nrf2 signaling may constitute a viable strategy to alleviate nanoparticle-induced respiratory injury.

Following the publication of the above article, an interested reader drew the authors' attention to the fact that, in Fig. 4B on p. 987, the 'Mon' and 'EBM84‑M' data panels showed apparently the same data, albeit the images were rotated through 180° relative to each other, suggesting that this figure part had been assembled incorrectly. The authors were able to consult their original data, and acknowledged that errors had been made in compiling Fig. 4B; specifically, the panel labeled as 'Mon' was inadvertently published using the image corresponding to the 'EBM84‑H' group; secondly, the panel labeled as 'EBM84‑M' incorrectly duplicated the published 'Mon' panel (which, as noted above, was the 'EBM84‑H' image); and finally, the panel labeled as 'EBM84‑H' mistakenly displayed the image corresponding to the 'EBM84‑M' group. The revised version of Fig. 4, now showing the correct data for the 'Mon', 'EBM84‑M' and 'EBM84‑H' data panels in Fig. 4B, is shown on the next page. The authors confirm that the errors made during the assembly of this figure did not have a significant impact on either the results or the conclusions reported in this study, and all the authors agree with the publication of this Corrigendum. The authors are grateful to the Editor of International Journal of Molecular Medicine for allowing them the opportunity to publish this Corrigendum; furthermore, they apologize to the readership of the Journal for any inconvenience caused. [International Journal of Molecular Medicine 31: 982‑988, 2013; DOI: 10.3892/ijmm.2013.1273].

IntroductionBoehmeria nivea (L.) Gaud. has traditionally been regarded as a medicinal food with applications in various inflammatory disorders. However, its role in chronic obstructive pulmonary disease (COPD) has not yet been clarified.MethodsIn this study, the preventive efficacy of the ethyl acetate fraction of B. nivea (L.) Gaud. leaves (EA-BN) was evaluated in a COPD model established by intratracheal instillation of lipopolysaccharide (LPS; 0.5 mg/kg body weight) and cigarette smoke condensate (CSC; 12.5 mg/kg body weight) in male C57BL/6N mice. The experimental groups received dexamethasone (3 mg/kg) as a positive control or EA-BN at doses of 100 and 200 mg/kg.ResultsEA-BN administration significantly reduced T helper 1 cytokine levels and decreased macrophage and neutrophil counts in bronchoalveolar lavage fluid. Histological analyses revealed that EA-BN mitigated alveolar destruction and inflammatory infiltration, whereas pulmonary function tests demonstrated improvements in the FEV0.1/FVC ratio and lung elastance in the LPS/CSC-induced COPD. Additionally, EA-BN alleviated oxidative stress by promoting the nuclear translocation of Nrf2 and enhancing the expression of its downstream targets, HO-1 and NQO1, leading to a reduction in reactive oxygen species and nitric oxide production. EA-BN downregulated thioredoxin-interacting protein and NLRP3 inflammasome activation, thereby suppressing caspase-1 and IL-1β expression, whereas also attenuating apoptosis by modulating the Bax/Bcl-2/caspase-3 pathway.DiscussionCollectively, these findings suggest that EA-BN possesses antioxidant, anti-inflammatory, and anti-apoptotic properties, supporting its potential as a preventive agent against COPD.

Copper oxide nanoparticles (CuONPs) are used in various industrial and commercial applications and are considered a potential hazard to humans and the environment. In particular, inhalation exposure to CuONPs causes severe damage to the respiratory tissue. Camellia sinensis L. is traditionally used to treat various inflammatory disorders and has been experimentally shown to exhibit anti-inflammatory and antioxidant properties due to its bioactive compounds. This study aimed to investigate the therapeutic effects of Camellia sinensis L. ethanolic extract (CSE) on pulmonary inflammation caused by CuONPs exposure, with an emphasis on the mitogen-activated protein kinase (MAPK) signaling pathway. The administration of CSE reduced the inflammatory cell counts including neutrophils and macrophages in mice exposed to CuONPs, as well as the levels of inflammatory cytokines, such as interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α. These events were accompanied by decreases in inflammatory cell infiltration into pulmonary tissue and mucus production from goblet cells. In addition, the administration of CSE reduced the activation of proteins involved in the MAPK pathway, including p-p38, p-JNK, p-ERK, and MUC5AC induced by CuONPs exposure. CSE effectively decreased the pulmonary inflammation caused by CuONPs exposure, which is associated with the suppression of the MAPK signaling pathway. Therefore, this evidence suggests that CSE has potential as a herbal remedy for controlling pulmonary inflammation caused by CuONPs exposure.

Allergic asthma is a widespread disease with elevated eosinophil levels. Although corticosteroids are widely prescribed for allergic asthma, numerous patients experience limited sensitivity and side effects. Loranthus tanakae Franch. & Sav., a traditional herbal plant, has anti-inflammatory and antioxidant properties in pulmonary inflammation caused by Asian sand dust and cigarette smoke condensate. To assess the protective effects of L. tanakae, we examined the influence of an L. tanakae ethanol extract (LTE) in ovalbumin (OVA)-induced asthma. Mice received intraperitoneal sensitisation with OVA, and challenged using OVA inhalation. LTE was consecutively orally gavaged for 6 days. Following sacrifice, the bronchoalveolar lavage fluid (BALF) and lung tissue was analysed. The LTE treatment considerably reduced inflammatory cell counts, proinflammatory cytokine levels in the BALF, and immunoglobulin E, compared with the OVA group, along with a reduction in airway hyperresponsiveness. The LTE also improved airway inflammation and suppressed mucus hypersecretion in the lung tissues. Additionally, the expression of MMP-9 and activation of ERK, JNK, and p38 were notably diminished in the LTE groups. This study revealed reduced airway inflammation in OVA-induced asthma via suppressing the MMP-9 and mitogen-activated protein kinase-associated factors. Consequently, our findings demonstrated that LTE is suggested as a potential remedy for allergic asthma.

Silibinin, a major compound of silymarin, has been reported to alleviate respiratory diseases including acute lung injury, asthma, chronic obstructive pulmonary disease, and pulmonary fibrosis through its antifibrotic, anti-inflammatory, and antioxidant properties. However, the specific mechanisms underlying its therapeutic effects, particularly in allergic asthma, are not fully understood. With the increasing prevalence and impact of allergic asthma, there is a need to elucidate the exact underlying mechanisms of its potential treatment effects. Herein, we investigated the therapeutic effects of silibinin on allergic asthma using house dust mite (HDM)-exposed mice and an HDM-stimulated human bronchial epithelium cell line, focusing on the roles of the NLR family CARD domain containing 4 (NLRC4) inflammasome and matrix metalloproteinase-9 (MMP-9). To induce airway inflammation, HDM extracts were instilled intranasally on days 0, 4, 8, and 12 to mice. Silibinin (20 and 40 mg/kg) was orally administered daily from days 0-12. The results showed that silibinin treatment attenuated allergic immune responses induced by HDM exposure, as evidenced by decreased airway hyperresponsiveness, reduced inflammatory cells and cytokines, lower immunoglobulin E levls, and decreased mucus production. Furthermore, silibinin treatment suppressed NLRC4 inflammasome activation and downregulated MMP-9 expression in the lungs. In HDM-stimulated cells, silibinin treatment decreased inflammatory cytokine levels and the expression of NLRC4 and interleukin-1β, indicating inhibition of NLRC4 inflammasome activation. Overall, our data demonstrated that silibinin alleviated allergic responses in HDM-induced asthmatic mice by inhibiting NLRC4 inflammasome activation and MMP-9 expression, underscoring its therapeutic potential in the treatment of asthma.

Background and Objectives: Acute kidney injury (AKI) is a common complication in patients with trauma and is associated with increased morbidity and mortality rates. Early identification of patients at risk of AKI may enable timely intervention and improved outcomes. Biomarkers such as urinary mitochondrial DNA copy number (mtDNAcn) may play a role in predicting AKI. However, its role as a predictor of AKI has rarely been studied in patients with trauma. Therefore, the aim of this study was to evaluate the utility of mtDNA for early detection of AKI in this patient population. Materials and Methods: This single-center prospective observational study included patients with trauma admitted to a regional trauma center between July 2022 and July 2023. Serum and urine samples were collected at baseline and at 24, 48, and 72 h to measure mtDNAcn using real-time polymerase chain reaction test. Clinical variables, including hemoglobin (Hb) levels, were also recorded. Results: Among 65 enrolled patients, 25 (38.5%) developed AKI. Patients with AKI showed significantly lower Hb levels and higher urinary mtDNAcn at admission. Multivariate logistic regression analysis identified low Hb and elevated urinary mtDNAcn as independent predictors of AKI. The optimal cutoff value was 10.95 g/dL for Hb and 738.0 copies/μL for urinary mtDNAcn. However, no significant temporal differences in serum mtDNAcn were observed between the AKI and no-AKI groups. Conclusions: Both Hb and urinary mtDNAcn may serve as independent biomarkers for early identification of AKI in patients with trauma. Future studies are warranted to determine optimal targets and validate these findings in larger multicenter cohorts.

BACKGROUND:Multi-organ distress syndrome (MODS) causes morbidity in patients with trauma. This study evaluates the effectiveness of interleukin-33 (IL-33), which reflects tissue damage and the inflammatory response, as a MODS indicator in patients with trauma. METHODS:Patients with trauma admitted to our trauma center between July 2022 and July 2023 were included. IL-33 levels were measured in blood samples for 4 days. Correlations with clinical and laboratory indicators, including initial IL-33 levels, were analyzed to identify independent predictors of MODS. RESULTS:Among the 87 patients enrolled, 20 developed MODS. Initial IL-33 levels were elevated in the MODS group, compared with the non-MODS group. In the non-MODS group, IL-33 levels increased on day 1 and then declined, whereas in the MODS group, IL-33 levels were highest at admission (day 0) and decreased continuously through day 3. In patients with detectable initial IL-33 levels, the measured levels correlated with higher Abbreviated Injury Scale 5 scores and the Injury Severity Score (ISS). A logistic regression analysis revealed the ISS and delta neutrophil index as factors contributing to MODS progression. CONCLUSIONS:The findings suggest that initial IL-33 levels are elevated in the MODS group, compared with non-MODS group, and exhibit a rapidly declining trend, showing an initial association with MODS that was not maintained in a multivariate analysis. These findings suggest that IL-33 might have relevance in assessing trauma severity; however, further validation is required before it can be considered a biomarker for MODS.

IMPORTANCE:Cryptorchidism, defined as the failure of one or both testes to descend into the scrotum, is a pathological condition that adversely impacts testicular development and spermatogenic function. In livestock, particularly bulls, such impairment can lead to substantial economic losses due to compromised fertility. A comprehensive understanding of the morphological and molecular alterations associated with cryptorchidism is essential for efforts to develop effective diagnostic and therapeutic strategies. OBJECTIVE:This study aimed to examine morphological changes and changes in the extracellular signal-regulated kinase (ERK) and cyclic AMP response element-binding protein (CREB) activity in the testes of bulls with cryptorchidism. METHODS:Testes from the 18-month-old unilateral cryptorchid bulls, including the descended (normal) and undescended (cryptorchid) testes from each animal, were analyzed through gross examination, histology, immunohistochemistry (Ki-67, phosphorylated p-ERK, p-CREB), and Western blotting. RESULTS:Cryptorchid testes were smaller and exhibited reduced epithelial thickness, although seminiferous tubule area remained unchanged. Expression of Ki-67, p-ERK, and p-CREB was significantly decreased, indicating impaired cell proliferation and signaling. Total ERK and CREB protein levels were unchanged, but phosphorylation levels were notably reduced in cryptorchid samples. CONCLUSIONS AND RELEVANCE:Our findings demonstrate that cryptorchidism alters testicular morphology and disrupts key signaling pathways, particularly those involved in cell growth and function. Such alterations may contribute to infertility and highlight the importance of early diagnosis and intervention in the context of reproductive management in livestock.

Copper oxide nanoparticles (CuONPs) are increasingly used across various industrial applications, raising concerns about their potential toxicity and necessitating comprehensive safety evaluations. In this study, we first evaluated the respiratory toxicity of CuONP exposure in a mouse model of asthma. CuONP exposure alone exacerbated asthma symptoms, as evidenced by increased airway hyperresponsiveness, inflammatory cell infiltration, and elevated cytokine production with increasing thioredoxin-interacting protein (TXNIP) expression. We then investigated whether TXNIP functions as a critical regulator of asthma exacerbation by examining the effects of TXNIP on the same experimental models using induced TXNIP-overexpressed mice induced by intratracheal injection of adeno-associated virus (AAV)2/8 and TXNIP knock-out (KO) mice. In CuONP-exposed asthmatic animals, TXNIP overexpressed animals significantly increased airway hyperresponsiveness, inflammatory cell counts and the production of helper type 2 cytokines, including interleukin (IL)-5 and IL-13, as well as the IL-1β, IL-6, tumor necrosis factor (TNF)-α, interferon gamma (IFN-γ) and IL-10 compared with the green fluorescent protein (GFP)-expressing controls. These responses were accompanied by the elevation of inflammatory infiltration, mucus production and the expression of apoptotic markers in lung tissues. By contrast, TXNIP KO animals markedly reduced the pathophysiological factors, inflammatory responses, mucus production and the expression of apoptotic markers in lung tissue compared with the wild-type (WT) animals. Collectively, these findings demonstrate that TXNIP plays a crucial role in regulating inflammatory responses and cell death in CuONP-exposed asthmatic mice, suggesting its potential as a therapeutic target for CuONP-induced asthma exacerbation.

Multi-organ dysfunction syndrome (MODS) is a life-threatening complication in trauma patients that is often triggered by a dysregulated immune response. We hypothesized that early dynamic changes in circulating cytokines, particularly interleukin-10 (IL-10), could help distinguish pathological immune responses leading to MODS from physiological responses associated with recovery. This study aimed to identify cytokine profiles predictive of MODS in the acute phase after trauma. Patients who were admitted to our regional trauma center between July 2022 and July 2023 were prospectively enrolled. Serum levels of interleukin (IL)-1β, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12, interferon gamma, and tumor necrosis factor alpha were quantified from blood samples collected over 3 consecutive days. Cytokines, clinical parameters, and other laboratory parameters were analyzed to identify independent predictors of MODS. Eighty-seven patients were included in this study, with 20 patients developing MODS. The Injury Severity Score (ISS), IL-6, and IL-10 levels were identified as significant predictors of MODS. The optimal cutoff values were 20.5 for ISS, 159.0 ρg/mL for IL-6, and 172.6 ρg/mL for IL-10. Temporal trends indicated that IL-6 and IL-10 levels peaked on day 1 post-injury and gradually declined by day 3. Serum levels of IL-6 and IL-10, along with the ISS, can serve as reliable biomarkers for predicting MODS in patients with trauma. While ISS is already a well-established tool in clinical practice, implementing IL-6 and IL-10 measurements will require developing cost-effective analytical techniques.

Ethnopharmacological relevance Boehmeria nivea (L.) Gaud. (BN) is a well-known traditional herbal medicine used to treat a wide range of conditions, including bleeding, fever, diarrhea, and wounds, as well as respiratory disorders such as cough and wheezing. However, its potential efficacy against allergic airway inflammation has yet to be systematically evaluated. Aim of the study This study aimed to investigate the therapeutic potential of BN, particularly the ethyl acetate fraction (EA-BN), in modulating immune responses in allergic asthma. Material and methods BALB/c mice were sensitized with ovalbumin (OVA) to induce allergic asthma and treated orally with BN fractions (100 mg/kg; EA-BN-L; 100 mg/kg and EA-BN-H; 200 mg/kg). Th2 cytokines (IL-4, IL-5, and IL-13) and inflammatory cell counts in bronchoalveolar lavage fluid (BALF) were measured. Bone marrow-derived dendritic cells (BMDCs) stimulated with lipopolysaccharide (LPS) and tumor necrosis factor (TNF)-α-stimulated NCI-H292 epithelial cells were used to assess in vitro potential effects. Flow cytometry and qPCR were used to evaluate cytokine levels and transcription factor expression. Results This study demonstrated the significant efficacy of EA-BN in an OVA-induced allergic asthma model. EA-BN treatment markedly suppressed the expression of extracellular cytokines (IL-10, IL-12, and TNF-α), co-stimulatory molecules (CD80 and CD86), and MHC class I/II in both LPS-stimulated BMDCs and splenic DCs in OVA-induced asthmatic mice. In addition, EA-BN reduced GATA-3 expression in CD3+CD4+ T cells, suggesting inhibition of Th2 cell differentiation in splenic T cells from asthmatic mice. Histological analysis revealed that EA-BN attenuated mucus overproduction, and inflammatory cell infiltration in lung tissues, along with a concurrent decrease in Th2 cytokine levels and MUC5AC expression. Furthermore, EA-BN also inhibited Th2 cytokine expression in TNF-α-stimulated NCI-H292 cells. Conclusion EA-BN mediates its immunoregulatory effects by simultaneously suppressing dendritic cell activation and blocking Th2 cell differentiation, ultimately leading to the downregulation of Th2-driven inflammatory responses. These findings support EA-BN as a potential herbal therapeutic candidate for allergic asthma.

Loranthus tanakae Franch. and Sav. is a traditional herbal remedy with anti-inflammatory and antioxidative properties, used to treat joint and respiratory inflammation. In this study, we investigated the therapeutic effects of L. tanakae ethanol extract (LTE) on atopic dermatitis (AD). An ovalbumin (OVA)-induced AD animal model and a human keratinocyte cell line, HaCaT, were used to assess LTE treatment effects on AD. An in vitro experiment showed that LTE treatment significantly decreased the production of regulated upon activation, normal T cell expressed and secreted (RANTES) cytokines and macrophage-derived chemokines (MDC) in tumor necrosis factor-alpha (TNF-α)/interferon-gamma (IFN-γ) (TNF-α/IFN-γ)-stimulated HaCaT cells in a concentration-dependent manner. In addition, treatment with LTE markedly reduced the translocation of signal transducer and activator transcription 1 (STAT1) protein to the nucleus and the phosphorylation of Janus kinase 2 (JAK2) in TNF-α/IFN-γ-stimulated HaCaT cells. In the in vivo experiment, administration of LTE significantly decreased the levels of immunoglobulin E (IgE) and interleukin-13 (IL-13) of OVA-induced AD mice, which was supported by histological evidence. Moreover, LTE treatment markedly reduced inflammatory cell infiltration and edema in the OVA-induced AD mice’s damaged lesions. In addition, applying LTE notably inhibited the phosphorylation of JAK2 and STAT1 in the OVA-induced AD mice, supported by in vitro results. In conclusion, LTE effectively alleviated the AD-induced skin inflammation in the OVA-induced AD animal model and TNF-α/IFN-γ-stimulated HaCaT cells; this was related to the suppression of JAK2 and STAT1 phosphorylation. These findings suggest that LTE has potential as a therapeutic agent for AD management.

Purpose Fournier gangrene (FG) is a life-threatening condition characterized by rapid tissue necrosis in the perineal and genital regions. Early identification of risk factors is essential for improving prognosis and reducing mortality rates. This study aimed to identify predictive factors associated with mortality in patients with FG. Methods A multi-institutional retrospective study was conducted across four tertiary care centers in Korea, including patients diagnosed with FG between January 2014 and December 2023. Data on demographic characteristics, laboratory findings, and clinical outcomes were collected. Independent risk factors for mortality were identified using multivariate logistic regression analysis, and optimal cutoff values for these predictors were determined. Results A total of 133 patients were included. The findings showed that age, initial lactate level, hemoglobin level, platelet count, and albumin level were significant predictors of mortality. Age was associated with an odds ratio of 1.105 (95% confidence interval [CI], 1.038–1.177; P=0.002), while initial lactate level exhibited an odds ratio of 1.820 (95% CI, 1.335–2.480; P<0.001). The optimal cutoff values identified were 64.5 years for age, 3.15 mmol/L for lactate, 8.65 g/dL for hemoglobin, 208×109/L for platelet count, and 3.05 g/dL for albumin, with varying sensitivity and specificity. Conclusion The study concluded that age, and initial levels of lactate, hemoglobin, albumin, and platelet count are independently predictive of mortality in patients with FG. These findings underscore the importance of aggressive management for patients presenting with abnormal serum values at admission to improve clinical outcomes. Further research is warranted to validate these results and to refine management strategies for FG.

This study investigated the median lethal dose and target organs of polyhexamethylene guanidine phosphate (PHMG-P)-an antimicrobial disinfectant-administered via single-dose intravenous injection (IV; 3.13, 6.25, or 12.5 mg/kg) and intratracheal instillation (ITI; 2, 4, or 8 mg/kg) in Sprague-Dawley rats. Alterations in serum biochemical (blood urea nitrogen, creatinine, and gamma-glutamyl transferase), hematological (reticulocyte count), and hepatic histopathological features, such as tubular dilation and atypical morphology, indicated that nephrotoxicity was induced after the 12.5 mg/kg IV. Changes in lung weight, hematological (neutrophil), and histopathological parameters, including alveolar macrophage aggregation, bronchiolar epithelium de-/regeneration, and granulomatous inflammation/pulmonary fibrosis, indicated that lung damage was induced after 8 mg/kg ITI. The acute IV LD50 was 12.20 and 12.83 mg/kg in male and female rats, respectively, and the target organ was the kidney. In contrast, the acute ITI LD50 was 5.68 and 5.01 mg/kg in male and female rats, respectively, and the target organ was the lung. This study indicates that PHMG-P exerts varying toxic effects depending on the route of administration.

Asian sand dust (ASD), generated from the deserts of China and Mongolia, mainly affects the human health of several countries in Northeast Asia including China, Korea, and Japan. In this study, we investigated the toxic effects of ASD on respiratory tract and explored the effects of ASD exposure on allergic asthma using ovalbumin-induced asthma model. C57BL/6 male mice were used for both the toxicity and allergic asthma studies. ASD (10, 20, and 40 mg/kg) was administered intranasally on days 1, 3, and 5. For allergic asthma, mice were sensitized with OVA (20 µg/mouse) and aluminum hydroxide (2 mg) on days 1 and 15, followed by OVA inhalation (1

Asian sand dust (ASD), also called China dust or yellow dust, mainly occurs in East Asia during spring and autumn. Because ASD enters the body mainly through the respiratory system, it can cause respiratory disorders or worsen underlying diseases. Because of this, it has become an important health concern that threatens the well-being of humans and animals. In this study, we investigated the effects of 15 and 30 mg/kg of Pycnogenol (PYC15 and 30 groups), a pine bark extract, on ASD-induced pulmonary inflammation in mice. We evaluated the inflammatory cell counts, inflammatory cytokines, and matrix-metalloproteinase (MMP)-9 expression in animal models. PYC administration significantly decreased inflammatory cell infiltration into lung tissue; this was accompanied by a reduction in the levels of proinflammatory mediators including interleukin (IL)-1β (P < 0.01), IL-6 (P < 0.01) and tumour necrosis factor-α (P < 0.01) in bronchoalveolar lavage fluids of ASD-exposed mice (ASD group). Histological analysis revealed that PYC suppressed ASD-induced pulmonary inflammation. Moreover, PYC suppressed the levels of matrix-metalloproteinase (MMP)-9 in the lung tissue of ASD-exposed mice, indicating that PYC reduced ASD-induced pulmonary inflammation by suppressing MMP-9. Together, these results indicate that PYC as the potential to treat ASD-driven pulmonary inflammation.

Ethnopharmacological relevance: Angelica decursiva Franchet & Savatier is a traditional medicinal plant used to treat asthma, cough, headache, pyrexia and thick phlegm in China, Japan and Korea. A. decursiva contains many types of coumarins, which can exert several pharmacological activities including anti-inflammatory and antioxidant properties for treating various diseases such as pneumonitis, atopic dermatitis, diabetes, and Alzheimer's disease.Aim of the study: In this study, we analyzed the components of A. decursiva ethanol extract (ADE) by high performance liquid chromatography (HPLC) and investigated the therapeutic effects of ADE against allergic asthma using lipopolysaccharide (LPS) stimulated RAW264.7 cells and an ovalbumin (OVA)-exposed allergic asthma model. To elucidate the mechanism of action of ADE, we examined the protein expression through network pharmacological analysis. Materials and methods: To establish asthma model, the mice were sensitized on day 0 and 14 via intraperitoneal injection of OVA with aluminum hydroxide. The mice were inhaled with OVA using an ultrasonic nebulizer on day 21, 22 and 23. ADE (50 and 100 mg/kg) was administered to mice by oral gave form day 18-23. On day 24, airway hyperresponsiveness (AHR) was measured using flexivent. On day 25, the mice were sacrificed and collected bronchoalveolar lavage fluids (BALF), serum and lung tissue. In LPS-stimulated RAW264.7 cell, nitric oxide and cytokines were measured. Additionally, expression of nuclear factor erythroid-2-related factor (Nrf2) and suppression of nuclear factor (NF)-?B were detected using double-immunofluorescence.Results: We detected the five coumarin components which included nodakenin, umbelliferon, (-)-marmesin (=nodakenetin), bergapten, and decursin, in ADE by high performance liquid chromatography. Treatment with ADE decreased the production of nitric oxide, interleukin (IL)-6 and tumor necrosis factor (TNF)-a in LPSstimulated RAW264.7 cells accompanied by the enhanced expression of nuclear factor erythroid-2-related factor (Nrf2) and suppression of nuclear factor (NF)-?B. In the asthma model, the administration of ADE reduced inflammatory cell count and airway hyperresponsiveness in OVA-exposed animals with decreased levels of IL-4, IL-13, and OVA-specific immunoglobulin E. These results were accompanied by the reduction of pulmonary inflammation and mucus secretion. Furthermore, ADE administration inhibited the expression of NF-?B and matrix metalloproteinase (MMP)-9 in OVA-exposed animals, which was consistent with the results of network pharmacological analysis.Conclusion: This study demonstrated that ADE effectively attenuated allergic inflammation induced by OVA inhalation through the enhancement of Nrf2 expression and suppression of NF-?B expression. Therefore, ADE may be a potential therapeutic agent for controlling asthma.