Comparison of anti-inflammatory efficacy between dexamethasone and a standardized herbal formula, PM014, in a cigarette smoke-induced subacute mouse model of chronic obstructive pulmonary disease
Kwan-Il Kim, Beom-Joon Lee
Division of Allergy, Immune and Respiratory System, Department of Internal Medicine, College of Korean Medicine, Kyung Hee University, KyungHeedae-ro, Dongdaemun-gu, Seoul 02447, Republic of Korea
|Date of Submission||04-Jun-2019|
|Date of Decision||10-Oct-2019|
|Date of Acceptance||19-Nov-2019|
|Date of Web Publication||15-Jun-2020|
Department of Internal Medicine (Pulmonary and Allergy System), Korean Medicine Hospital, 23, Kyung Hee Dae-ro, Dongdaemun-gu, Seoul 130-872
Republic of Korea
Source of Support: None, Conflict of Interest: None
| Abstract|| |
Objective: A standardized herbal formula, PM014, originated from Chungsangboha -tang, which has been used to treat various respiratory diseases, including bronchitis, asthma, and emphysema. Several previous studies have reported that the therapeutic mechanism of PM014 was mediated by an anti-inflammatory effect. Therefore, we compared anti-inflammatory efficacy between PM014 and dexamethasone (DEX) using a mouse model of cigarette smoke (CS)-induced subacute pulmonary inflammation. Materials and Methods: Female C57BL/6 mice were revealed to CS for 2 h/day, three cigarettes per day, 5 days a week for 3 weeks; the control group got no other treatment, while the DEX and PM014 groups received 1 mg/kg of DEX and 100 mg/kg of PM014, respectively (both were orally administered). The histological morphology and average alveolar size were determined by lung histology. The inflammatory cell profiles and protein expressions of pro-inflammatory cytokines including tumor necrosis factor-alpha (TNF-α), interleukin (IL)-1 β, IL-6, and one pro-inflammatory chemokine (monocyte chemoattractant protein 1 [MCP-1]), were measured in bronchoalveolar lavage fluid (BALF). The mRNA expressions of the pro-inflammatory cytokines and chemokine were also measured in lung tissues. Results: Both PM014 and DEX attenuated histological injury and air space enlargement in lung tissue and decreased the number of inflammatory cells in BALF. Both also decreased the mRNA expressions of TNF-α, IL-6, and IL-1 β in lung tissue and reduced the protein expressions of TNF-α, IL-1 β, IL-6, and MCP-1 in BALF. Conclusion: Our results showed that the anti-inflammatory efficacy of PM014 and DEX was equivalent in a subacute mouse model of CS-induced chronic obstructive pulmonary disease.
Keywords: Anti-inflammation, chronic obstructive pulmonary disease, cigarette smoke, corticosteroid, PM014
|How to cite this article:|
Kim KI, Lee BJ. Comparison of anti-inflammatory efficacy between dexamethasone and a standardized herbal formula, PM014, in a cigarette smoke-induced subacute mouse model of chronic obstructive pulmonary disease. Phcog Mag 2020;16:321-8
|How to cite this URL:|
Kim KI, Lee BJ. Comparison of anti-inflammatory efficacy between dexamethasone and a standardized herbal formula, PM014, in a cigarette smoke-induced subacute mouse model of chronic obstructive pulmonary disease. Phcog Mag [serial online] 2020 [cited 2021 Sep 23];16:321-8. Available from: http://www.phcog.com/text.asp?2020/16/69/321/286733
- PM014, originated from Chungsangboha -tang, which has been used to treat various respiratory diseases, demonstrated anti-inflammatory efficacy similar to that of dexamethasone in a mouse model of subacute cigarette smoke-induced chronic obstructive pulmonary disease (COPD). The results of the present study suggest that PM014 could be used as an adjunctive agent or substitute for oral steroids in COPD patients.
Abbreviations used: BALF; bronchoalveolar lavage fluid, CS; cigarette smoke, DEX; dexamethasone, IL; interleukin, MCP; monocyte chemoattractant protein, TNF-α; tumor necrosis factor-alpha
| Introduction|| |
Cigarette smoking is one of the major global health problems, which has been the cause of more than 1 in 10 deaths. It mainly causes respiratory diseases, cardiovascular diseases, and cancers., Cigarette smoke (CS) is a complex mixture of various noxious chemical compounds, such as nicotine, tar, and benzopyrene; this mixture can rapidly induce inflammation and injury of the tissues, common pathologic features, in the CS-associated diseases. When CS enters the lungs, it activates the immune-inflammatory pathway, thus increasing the activity of macrophages, neutrophils, and lymphocytes, as well as the production of related mediators (e.g., pro-inflammatory cytokines such as tumor necrosis factor-alpha [TNF-α], interleukin (IL)-1 β and IL-6 and chemokines such as monocyte chemoattractant protein 1 [MCP-1]). This inflammation can induce chronic obstructive pulmonary disease (COPD)-like lung damage, as well as the destruction and obstruction of various parts of the airways.,,
Glucocorticoids have been commonly used for the treatment of many airway inflammatory diseases and are known as typical anti-inflammatory therapeutic agents., Notably, glucocorticoid was shown to have a preventive effect against COPD in the TORCH study. However, glucocorticoid can cause a variety of side effects, including weight gain, Cushing syndrome, muscle breakdown, insulin resistance, osteoporosis, mood changes, and glaucoma. Therefore, as alternatives to glucocorticoids, other agents with anti-inflammatory effects have been developed for the various respiratory diseases.
A standardized herbal formula, PM014, originated from Chungsangboha -tang, which has been used to treat chronic respiratory disease (e.g., asthma and COPD) in Korea for centuries. The original herbal formula contained 18 species of medical plants, leading to difficulties in standardization; thus, it was simplified to contain seven herbs and renamed PM014. Previous studies have shown the therapeutic effects of PM014 on various respiratory diseases through its anti-inflammatory effects.,,, We aimed to confirm the preventive effect of PM014 against inflammation and to compare anti-inflammatory efficacy between PM014 and dexamethasone (DEX), through histological analysis and measuring the expression of cytokines and chemokines in lung tissues and bronchoalveolar lavage fluid (BALF) using a mouse model of subacute CS-induced COPD.
| Materials and Methods|| |
The methods of extraction and standardization of PM014 were described in a previous report. Seven medicinal herbs were used to prepare PM014 [Table 1]. Each extract granule was purchased from Sun Ten Pharmaceutical (Taipei, Taiwan), and mixtures of the herbal extract granules were dissolved in 0.9% saline to achieve a final concentration of 10% (w/v). A supernatant was then obtained by centrifugation at 27,000 ×g for 20 min at 4°C (Eppendorf, Hamburg, Germany) and sterilized by passage through a 0.22-μm syringe filter. The stock of the PM014 extract was then diluted and stored at 4°C. DEX (Sigma Aldrich, St. Louis, MO, USA), which was used as a positive control, was also dissolved in saline.
Female C57BL/6 mice, 6–7 weeks old and weighing 16–17 g, were obtained from Orient Bio Inc., (Seongnam, South Korea). Mice were housed in sterile microisolator cages and kept at 20°C under a 12-h day–night cycle and fed a standard sterile diet with free access to water. Bodyweight of each mouse was measured individually every 2 days using a digital scale with 0.1 g accuracy and adjusting animal movement function (Ohaus Corporation, Parsippany, NJ, USA). All experiments were conducted in compliance with the requirements of the Animal Care and Ethics Committee of Kyung Hee University. Twenty mice were divided into four groups. In the normal group (NOR), mice were freely disclosed to room air within a chamber and treated with distilled water (DW). The other three groups were the control (CON), DEX, and PM014 groups; all mice in these groups were exposed to CS. CS was made from the standard reference cigarettes 3R4F (University of Kentucky, Lexington, KY, USA), using a smoking apparatus. Mice were exposed to CS from three cigarettes for 2 h, 5 days/week for 3 weeks. The 2 h of exposure were composed of 30 min exposure to fresh air between 1-h exposure to CS. The CON was treated with DW, the DEX group with 1 mg/kg DEX, and the PM014 group with 100 mg/kg PM014. DW, DEX, and PM014 were administered orally once per day for 3 weeks following the first CS exposure. All mice were sacrificed on day 22 and their lung tissues were extracted [Figure 1].
|Figure 1: Schedule for cigarette smoke exposure. Twenty female C57BL/6 mice were divided into four groups (n = 5 per group): normal group (air + distilled water), control (cigarette smoke + distilled water), dexamethasone (cigarette smoke + dexamethasone), and PM014 (cigarette smoke + PM014). The mice were exposed to the smoke from three cigarettes for 2 h/day, 5 days/week for 3 weeks|
Click here to view
Collection of bronchoalveolar lavage fluid
Bronchoalveolar lavage was performed by cannulating the trachea and washing the lungs three times with phosphate-buffered saline (PBS; pH, 7.2; room temperature), using a volume of 700 mL/wash. The retrieved fluid was stored on ice. After washing three times, the BALF was centrifuged for 10 min at 1,300 rpm. The concentrations of cells were measured using a hemocytometer. After cytocentrifugation and Diff-Quick staining, differential cell counts were conducted. A total of approximately 500 cells were counted.
The cell-free supernatant was stored at −80°C until further use.
RNA extraction and real-time polymerase chain reaction
To measure the gene expression of TNF-α, IL-1 β, IL-6, and MCP-1, total RNA from whole lung tissues was isolated using TRIzol reagent (Invitrogen, Carlsbad, CA, USA) in accordance with the manufacturer's protocol. 2 μg of RNA were reverse-transcribed using random hexamer primers and M-MuLV reverse transcriptase (Fermentas, Pittsburgh, PA, USA). The synthesized complementary DNA was used for polymerase chain reaction (PCR) amplification, and real-time PCR was performed. The Thermal Cycler Dice Real-Time PCR System (Takara Bio Inc., Shiga, Japan) was used according to the protocol. The forward and the reverse primers (Cosmogenetech, Seoul, Korea) were as follows: The primers of TNF-α were 5'-CAAGGGACAAGGCTGCCCG-3' and 5'-TAGACCTGCCCGGACTCCGC-3'; The primers of IL-1β were 5'-TCATGGGATGATGATGATAACCTGCT-3' and 5'-CCCATACTTTAGGAAGACACGGATT-3'; The primers of IL-6were 5'-TGCTCCTGACAACCACGGCCT-3' and 5'-ACAGGTCTGTTGGGAGTGGTATCCT-3'; The primers of MCP-1 were5'-TCACAGTTGCCGGCTGGAGC-3' and 5'-CAGCAGGTGAGTGGGGCGTT-3'; The primers of GAPDH were 5'-TCTGACGTGCCGCCTGGAGA-3' and 5'-TGGGCCCTCAGAYGCCTGCT-3'. A single peak for each reaction for TNF-α, IL-1 β, IL-6, MCP-1, and GAPDH was confirmed by dissociation curve analyses, and 40 cycles of PCR were performed under the following cycle conditions, which were denaturation for 10 s at 95°C, annealing for 10 s at 60°C, and elongation for 12 s at 72°C. After the threshold of the gene of interest was measured three times, the mean cycle threshold was calculated and normalized with the mean cycle threshold of the internal reference gene, GAPDH.
Enzyme-linked immunosorbent assay
According to the manufacturer's instructions, TNF-α, IL-1 β, IL-6, and MCP-1 were measured by an enzyme-linked immunosorbent assay (ELISA) kit (OptELA™ Kits; BD Biosciences, San Diego, CA, USA). In brief, 96-well plates were prepared and coated for 24 h at 4°C with a coating buffer. The buffer contained the antibodies of TNF-α, IL-1 β, IL-6, or MCP-1 as capture antibodies. The plates were washed three times using PBS with Tween-20 (PBS-T20), respectively. They were blocked for 1 h at room temperature using PBS with 10% fetal bovine serum and then washed with PBS-T20. Following to add the standard to the wells with diluted culture media and the plates were incubated for 2 h. After washing, the detection antibodies of TNF-α, IL-1 β, IL-6, and MCP-1 were added to the wells. Subsequently, the plates were incubated for 1 h and washed seven times. Then, the TMB Substrate Reagent (BD Biosciences, San Diego, CA, USA) was added and incubated for 30 min at room temperature. After the reaction was induced, the stop solution (2N H2 SO4) was added for fixation. The optical density of each well was read on an ELISA plate reader using 450- and 570-nm filters.
Morphometric and histological analysis
The lung was harvested, and the left lung was inflated with fixatives followed by paraffin embedding. For histological examination, the lung was cut in 4-μm thick slices using a rotary microtome (Leica, Nussloch, Germany) and stained with hematoxylin and eosin (H and E). For the quantitating airspace enlargement, the maximum cross-section of parenchyma was selected and the morphometric assessment was conducted using digital image tool. The mean alveolar airspace area was calculated from the sum of the lumen divided by the number of alveoli by Image Pro-plus 5.1 software (Media Cybernetics, Silver Spring, MD, USA).
All data were statistically analyzed using Prism 5 software (GraphPad Software Inc., La Jolla, CA, USA) and are expressed as mean ± standard error of the mean. One-way ANOVA followed by Newman–Keuls post hoc test was used for multiple comparisons. P < 0.05 was considered statistically significant.
| Results|| |
Effects of PM014 and dexamethasone on histological damage and air space enlargement in lung tissue
To determine the effects of PM014 and DEX on lung tissue damage caused by CS exposure, we performed a histological examination using H and E staining. Airway space area enlargement meaning the destruction of the lung parenchyma was measured as the mean alveolar air spaces area (μm). The CON group showed significantly greater airspace enlargement compared with the NOR group, while PM014 and DEX groups showed significantly less airspace enlargement – to a similar extent – compared with the CON group [Figure 2]. These results mean that PM014 and DEX inhibited CS-induced pulmonary alveolar destruction to a similar degree.
|Figure 2: Effects of PM014 and dexamethasone on the histological morphology of lung tissue. Lung tissues were fixed, sectioned at a thickness of 4 μM, and stained with hematoxylin and eosin (H and E solution; magnification, ×100) (a). The evaluation of airspace changes by digital image analysis of H and E stained sections (b). Values are expressed as mean ± standard error of the mean. Statistical analysis was performed using one-way ANOVA followed by Newman–Keuls post hoc test. **P < 0.01, ***P < 0.001|
Click here to view
Effects of PM014 and dexamethasone on the inflammatory cell count in bronchoalveolar lavage fluid
Because macrophages and neutrophils play an important role in smoking-induced COPD, we investigated their counts in BALF. We found that the total cell, neutrophil, and macrophage counts were significantly higher in the CS group than in the CON group, whereas they were significantly lower in the PM014 and DEX groups than in the CS group [Figure 3]a, [Figure 3]b, [Figure 3]c. These results indicated that PM014 exerted anti-inflammatory effects in CS-induced COPD-like lung inflammation, which were similar to those observed in the DEX group.
|Figure 3: Effects of PM014 and dexamethasone on the inflammatory cell profiles in bronchoalveolar lavage fluid. The numbers of inflammatory cells in bronchoalveolar lavage fluid were measured after 3 weeks of cigarette smoke exposure. The total cell (a), macrophage (b), and neutrophil (c) counts were determined in bronchoalveolar lavage fluid using light microscopy after Diff-Quick staining. Values are expressed as mean ± standard error of the mean. All statistical analyses were performed using one-way ANOVA followed by Newman–Keuls post hoc test. ***P < 0.001|
Click here to view
Effects of PM014 and dexamethasone on mRNA expression of pro-inflammatory cytokines and monocyte chemoattractant protein 1 in lung tissues
To confirm the anti-inflammatory effects of PM014 and DEX, we evaluated mRNA expression in whole lung tissues using real-time PCR. Because TNF-α, IL-1 β, IL-6, and MCP-1 are common inflammatory mediators in the lung, the mRNA expression levels of these mediators were evaluated the degree of pulmonary inflammation. Notably, the mRNA expression levels of TNF-α, IL-1 β, IL-6, and MCP-1 increased significantly in lung tissues from the CS group compared with tissues from the CON group. In contrast, the tissues from the PM014 group showed significantly lower levels of TNF-α, IL-6, and IL-1 β relative to those in the CS group. Tissues from the DEX group were used as positive controls; these showed significant reductions in the inflammatory mediators, similar to the PM014 group [Figure 4].
|Figure 4: Effects of PM014 and dexamethasone on the mRNA expression of pro-inflammatory cytokines and monocyte chemoattractant protein 1 in lung tissues. mRNA levels of tumor necrosis factor-alpha (a), interleukin-6 (b), interleukin-1 β (c), and monocyte chemoattractant protein 1 (d) were measured by a real-time polymerase chain reaction in the lung tissues. The results are shown as fold changes in expression relative to GAPDH, a reference gene. Values are expressed as mean ± standard error of the mean. Statistical analysis was performed using one-way ANOVA followed by Newman–Keuls post hoc test. *P < 0.05, **P < 0.01, ***P < 0.001|
Click here to view
Effects of PM014 and dexamethasone on the release of pro-inflammatory cytokines and monocyte chemoattractant protein 1 in bronchoalveolar lavage fluid
To determine the anti-inflammatory effects of PM014 and DEX at the protein level, ELISA analysis was performed on BALF from CS-exposed mice. This analysis revealed that significantly higher levels of TNF-α, IL-1 β, IL-6, and MCP-1 protein were released in the CS group than in the CON group. A number of pro-inflammatory mediator proteins showed reductions in the PM014 group. In addition, PM014 decreased significantly the protein expressions of TNF-α, IL-1 β, IL-6, and MCP-1 compared with CS. However, the degree of reduction was similar between PM014 and DEX [Figure 5].
|Figure 5: Effects of PM014 and dexamethasone on protein levels of pro-inflammatory cytokines and monocyte chemoattractant protein 1 in bronchoalveolar lavage fluid. Protein levels of tumor necrosis factor-alpha (a), interleukin-6 (b), interleukin-1 β (c), and monocyte chemoattractant protein 1 (d) in bronchoalveolar lavage fluid were measured by enzyme-linked immunosorbent assay. Values are expressed as mean ± standard error of the mean. Statistical analysis was performed using one-way ANOVA followed by Newman–Keuls post hoc test. *P < 0.05, **P < 0.01, ***P < 0.001|
Click here to view
| Discussion|| |
In the current subacute mouse model of CS-induced COPD, anti-inflammatory effects in groups treated with 100 mg/kg of PM014 were similar to those observed in groups treated with 1 mg/kg of DEX. PM014 reduced the total cell, macrophage and neutrophil numbers in BALF, inhibited lung injury and reduced mRNA levels and protein levels of pro-inflammatory cytokines (TNF-α, IL-1β and IL-6) and a pro-inflammatory chemokine (MCP-1), similar to DEX.
CS exposure causes an inflammatory response in the lungs, which results in a variety of chronic lung disorders, including COPD. The most important mechanism in the inflammatory response is the collection of numerous innate and adaptive immune cells from the blood; these include leukocytes, lymphocytes (neutrophils), macrophages, and dendritic cells. The inflammatory cells infiltrate the airways and lung parenchyma, eventually inducing emphysematous destruction of the structure and function of alveoli. In the present study, 3 weeks of CS exposure-induced COPD-like lung injuries, including airway destruction and increased inflammatory cells, as in previous studies.,,
Since COPD was first identified approximately 10 years ago, oral corticosteroids have been widely used to control it., However, long-term oral steroid treatment for stable COPD has been reported to cause a deterioration of respiratory and peripheral muscle strength and pulmonary function, as well as comorbidities, such as diabetes, hypertension, and osteoporosis, and increased mortality; thus, the oral corticosteroid is recommended for use only in patients with an acute exacerbation of COPD. Accordingly, there has been a demand for the development of other anti-inflammatory medicines that could substitute for, or enhance treatment with, inhaled steroid. The anti-leukotriene agent, montelukast, as a representative example, is widely used and has shown anti-inflammatory efficacy similar to that of DEX. Notably, DEX is a synthetic glucocorticoid that has shown an anti-inflammatory effect. It was reported to inhibit release of granulocyte-macrophage colony-stimulating factor from monocytes, U937 monocytic cells, and porcine alveolar macrophages; it was also reported to decreasein vitro release of IL-8, a chemokine for attraction and activation of neutrophils, from human airway smooth muscle cells and human airway epithelial cells. Furthermore, DEX has been shown to induce transcription of IκBα, the cytoplasmic inhibitor of NF-κB,, thereby suppressing the expression of pro-inflammatory cytokines, including TNF-α, IL-1, IL-6, and IL-8, and cell adhesion molecules involved in the migration of monocytes into the extravascular space, including MCP-1. Based on these mechanisms, DEX has been used as a positive control in various anti-inflammatory studies involving CS exposure.,,
The anti-inflammatory effect of PM014 has been reported in various lung inflammation models.,,, A standardized herbal formula, PM014, contains seven species of medicinal plants, some of which have been studied for their anti-inflammatory effects. Hong et al . Showed that Paeonia suffruticosa inhibited inflammatory responses by regulating the nuclear factor-κB/IκB signaling pathway. The roots of Stemonae sessilifolia have long been used to relieve cough, tuberculosis, and bronchitis; the main alkaloids of this plant have shown antitussive activity in a guinea pig cough model.Rehmannia glutinosa has an effect on nitric oxide inhibitory activities. The major components of Scutellaria baicalensis include baicalin, baicalein, and wogonin, which are bioactive flavones. Baicalin has been reported to ease inflammation in a model of CS-induced inflammation., The effects of PM014 and its individual constituent herbs on lung inflammation were investigated in a previous study, in which PM014 elicited the reduction of immune cell recruitments in the lungs compared with individual herbs.
Histological changes associated with COPD include enlargement of alveolar airspaces due to destruction of alveolar septa, as well as an increase in the number of airspaces. Thus, airspace enlargement is used as a criterion for determining the severity of emphysematous changes., In the present study, PM014 attenuated CS-induced airspace enlargement to a similar extent compared to DEX, which suggests that both agents prevented alveolar destruction in a similar manner.
Macrophages and neutrophils play important roles in CS-induced lung inflammation. Macrophages, the first line of defense of the lung against foreign bodies, mediate the inflammatory process through the release of chemokines that attract various immune cells, including neutrophils., Neutrophils migrate to the airway and are activated by chemotactic factors., Then activated neutrophils and macrophages cause lung destruction by secretion of various proteolytic enzymes, such as neutrophil elastase, matrix metalloproteinases (MMPs), including MMP-8, MMP-9, and MMP-12 (macrophage elastase), and numerous lung-damaging pro-inflammatory cytokines and chemokines.,, There have been several reports showing that increasing macrophages and neutrophils in BALF or sputum were correlated with COPD severity in human and murine models.,,, In the present study, PM014 and DEX showed similar effects in reducing macrophages and neutrophils in BALF.
Various cytokines play crucial roles in the pathological progression of COPD. TNF-α, IL-1β, IL-6, and MCP-1 have been suggested as underlying causes of lung inflammation and COPD development. With respect to the relationships between these cytokines and CS, the levels of TNF-α, IL-1 β, IL-6, IL-8, and MCP-1 are known to be increased in the BALF of chronic smokers. TNF-α is an important chemotactic protein for neutrophils; it may activate airway inflammatory cells to release inflammatory mediators through the upregulation of adhesion-molecule expression, thus enhancing the migration of inflammatory cells into the lower airways and activating profibrotic mechanisms involved in airway remodeling. TNF-α is also believed to play a key role in CS-induced lung inflammation; notably, CS exposure induces TNF-α overexpression in lung tissue, which stimulates the secretion of MMPs from macrophages, resulting in an emphysematous change in the alveoli. Clinical analyses of COPD patients have shown that TNF-α levels are increased in sputum and blood. IL-1 β is another important pro-inflammatory cytokine in COPD, and CS-induced emphysematous changes increased IL-1 β has been observed in the BALF and sputum of smokers, and is inversely correlated with pulmonary function in COPD patients, but directly correlated with the severity of COPD. IL-6 is a pleiotropic cytokine that regulates the immune system and has been associated with the progression of COPD severity; increased serum IL-6 has been reported in COPD patients and was a predictive factor of mortality in COPD., Monocyte recruitment to the lungs is an important step in the progression of COPD: monocytes release many macromolecules that mediate inflammation. In addition, MCP-1 is an important chemokine that regulates migration and infiltration of monocytes and macrophages; levels of MCP-1 were found to be increased in the BALF of COPD patients and the sputum of smokers. The above pro-inflammatory cytokines influence each other during the progression of inflammation in COPD., In particular, it is well-known that IL-1 β and TNF-α promote the production of each other; this is because their signaling pathways are similar. Therefore, IL-1 β induces the production of TNF-α from alveolar macrophages and the CS-induced increase in TNF-α is blocked by the inhibition of IL-1 β. In addition, pro-inflammatory agonists induce increases in IL-1 β and IL-6, and IL-6 and MCP-1 are released concomitantly from the monocytes of COPD patients on stimulation with lipopolysaccharide. Therefore, blockade of various inflammatory pathways and mediators, including inhibition of the cytokines and chemokines assessed in the present study, may constitute an ideal method to prevent COPD. In the present study, PM014 and DEX induced similar reductions in the protein and mRNA expression levels of pro-inflammatory cytokines and chemokines.
| Conclusion|| |
In the present study, PM014 demonstrated anti-inflammatory efficacy similar to that of DEX in a mouse model of subacute CS-induced COPD. Previous studies have shown that PM014 is effective in various lung inflammation models; the results of the present study suggest that PM014 could be used as an adjunctive agent or substitute for oral steroids in COPD patients. Future studies should include assessments of underlying molecular mechanisms; furthermore, clinical studies of side effects and potential interactions with inhaled steroids are needed.
Authors are thankful to Korea Health Industry Development Institute, funded by the Ministry of Health and Welfare, Republic of Korea.
Financial support and sponsorship
This research was supported by a grant from the Korea Health Technology R and D Project through the Korea Health Industry Development Institute, funded by the Ministry of Health and Welfare, Republic of Korea (HI18C1944).
Conflicts of interest
There are no conflicts of interest.
| References|| |
Reitsma MB, Fullman N, Ng M, Salama JS, Abajobir A, Abate KH, et al
. Smoking prevalence and attributable disease burden in 195 countries and territories, 1990-2015: A systematic analysis from the Global Burden of Disease Study 2015. Lancet 2017;389:1885-906.
Sopori M. Effects of cigarette smoke on the immune system. Nat Rev Immunol 2002;2:372-7.
Stämpfli MR, Anderson GP. How cigarette smoke skews immune responses to promote infection, lung disease and cancer. Nat Rev Immunol 2009;9:377-84.
Bhalla DK, Hirata F, Rishi AK, Gairola CG. Cigarette smoke, inflammation, and lung injury: A mechanistic perspective. J Toxicol Environ Health B Crit Rev 2009;12:45-64.
Brusselle GG, Joos GF, Bracke KR. New insights into the immunology of chronic obstructive pulmonary disease. Lancet 2011;378:1015-26.
Nikota JK, Stämpfli MR. Cigarette smoke-induced inflammation and respiratory host defense: Insights from animal models. Pulm Pharmacol Ther 2012;25:257-62.
Barnes PJ. Anti-inflammatory actions of glucocorticoids: Molecular mechanisms. Clin Sci (Lond) 1998;94:557-72.
Lannan EA, Galliher-Beckley AJ, Scoltock AB, Cidlowski JA. Proinflammatory actions of glucocorticoids: Glucocorticoids and TNFα coregulate gene expression in vitro
and in vivo
. Endocrinology 2012;153:3701-12.
Celli BR, Thomas NE, Anderson JA, Ferguson GT, Jenkins CR, Jones PW, et al
. Effect of pharmacotherapy on rate of decline of lung function in chronic obstructive pulmonary disease: Results from the TORCH study. Am J Respir Crit Care Med 2008;178:332-8.
Schäcke H, Döcke WD, Asadullah K. Mechanisms involved in the side effects of glucocorticoids. Pharmacol Ther 2002;96:23-43.
Roh GS, Seo SW, Yeo S, Lee JM, Choi JW, Kim E, et al
. Efficacy of a traditional Korean medicine, Chung-Sang-Bo-Ha-Tang, in a murine model of chronic asthma. Int Immunopharmacol 2005;5:427-36.
Lee H, Kim Y, Kim HJ, Park S, Jang YP, Jung S, et al
. Herbal formula, PM014, attenuates lung inflammation in a murine model of chronic obstructive pulmonary disease. Evid Based Complement Alternat Med 2012;2012:769830.
Jung KH, Haam KK, Park S, Kim Y, Lee SR, Lee G, et al
. The standardized herbal formula, PM014, ameliorated cigarette smoke-induced lung inflammation in a murine model of chronic obstructive pulmonary disease. BMC Complement Altern Med 2013;13:219.
Jung KH, Choi HL, Park S, Lee G, Kim M, Min JK, et al
. The effects of the standardized herbal formula PM014 on pulmonary inflammation and airway responsiveness in a murine model of cockroach allergen-induced asthma. J Ethnopharmacol 2014;155:113-22.
Kim JY, Shin D, Lee G, Kim JM, Kim D, An YM, et al
. Standardized Herbal Formula PM014 Inhibits Radiation-Induced Pulmonary Inflammation in Mice. Sci Rep 2017;7:45001.
Epaud R, Aubey F, Xu J, Chaker Z, Clemessy M, Dautin A, et al
. Knockout of insulin-like growth factor-1 receptor impairs distal lung morphogenesis. PLoS One 2012;7:e48071.
Leberl M, Kratzer A, Taraseviciene-Stewart L. Tobacco smoke induced COPD/emphysema in the animal model-are we all on the same page? Front Physiol 2013;4:91.
Decramer M, Janssens W, Miravitlles M. Chronic obstructive pulmonary disease. Lancet 2012;379:1341-51.
Pauwels NS, Bracke KR, Dupont LL, Van Pottelberge GR, Provoost S, Vanden Berghe T, et al
. Role of IL-1α and the Nlrp3/caspase-1/IL-1β axis in cigarette smoke-induced pulmonary inflammation and COPD. Eur Respir J 2011;38:1019-28.
Balamayooran G, Batra S, Cai S, Mei J, Worthen GS, Penn AL, et al
. Role of CXCL5 in leukocyte recruitment to the lungs during secondhand smoke exposure. Am J Respir Cell Mol Biol 2012;47:104-11.
Lee E, Yun N, Jang YP, Kim J. Lilium lancifolium
Thunb. extract attenuates pulmonary inflammation and air space enlargement in a cigarette smoke-exposed mouse model. J Ethnopharmacol 2013;149:148-56.
Callahan CM, Dittus RS, Katz BP. Oral corticosteroid therapy for patients with stable chronic obstructive pulmonary disease. A meta-analysis. Ann Intern Med 1991;114:216-23.
Standards for the diagnosis and care of patients with chronic obstructive pulmonary disease. American Thoracic Society. Am J Respir Crit Care Med 1995;152:S77-121.
Walters JA, Walters EH, Wood-Baker R. Oral corticosteroids for stable chronic obstructive pulmonary disease. Cochrane Database Syst Rev 2005;20:CD005374.
Horita N, Miyazawa N, Morita S, Kojima R, Inoue M, Ishigatsubo Y, et al
. Evidence suggesting that oral corticosteroids increase mortality in stable chronic obstructive pulmonary disease. Respir Res 2014;15:37.
Vogelmeier CF, Criner GJ, Martinez FJ, Anzueto A, Barnes PJ, Bourbeau J, et al
. Global strategy for the diagnosis, management, and prevention of chronic obstructive lung disease 2017 report. GOLD Executive Summary. Am J Respir Crit Care Med 2017;195:557-82.
Basyigit I, Sahin M, Sahin D, Yildiz F, Boyaci H, Sirvanci S, et al
. Anti-inflammatory effects of montelukast on smoke-induced lung injury in rats. Multidiscip Respir Med 2010;5:92-8.
Abdel Kawy HS. Montelukast versus dexamethasone treatment in a guinea pig model of chronic pulmonary neutrophilic inflammation. COPD 2016;13:455-63.
Seldon PM, Stevens DA, Adcock IM, O'Connor BJ, Barnes PJ, Giembycz MA. Albuterol does not antagonize the inhibitory effect of dexamethasone on monocyte cytokine release. Am J Respir Crit Care Med 1998;157:803-9.
Deaton PR, McKellar CT, Culbreth R, Veal CF, Cooper JA Jr., Hyperoxia stimulates interleukin-8 release from alveolar macrophages and U937 cells: Attenuation by dexamethasone. Am J Physiol 1994;267:L187-92.
Lin G, Pearson AE, Scamurra RW, Zhou Y, Baarsch MJ, Weiss DJ, et al
. Regulation of interleukin-8 expression in porcine alveolar macrophages by bacterial lipopolysaccharide. J Biol Chem 1994;269:77-85.
John M, Au BT, Jose PJ, Lim S, Saunders M, Barnes PJ, et al
. Expression and release of interleukin-8 by human airway smooth muscle cells: Inhibition by Th-2 cytokines and corticosteroids. Am J Respir Cell Mol Biol 1998;18:84-90.
Kwon OJ, Au BT, Collins PD, Baraniuk JN, Adcock IM, Chung KF, et al
. Inhibition of interleukin-8 expression by dexamethasone in human cultured airway epithelial cells. Immunology 1994;81:389-94.
Scheinman RI, Cogswell PC, Lofquist AK, Baldwin AS Jr. Role of transcriptional activation of I kappa B alpha in mediation of immunosuppression by glucocorticoids. Science 1995;270:283-6.
Auphan N, DiDonato JA, Rosette C, Helmberg A, Karin M. Immunosuppression by glucocorticoids: Inhibition of NF-kappa B activity through induction of I kappa B synthesis. Science 1995;270:286-90.
Jantz MA, Sahn SA. Corticosteroids in acute respiratory failure. Am J Respir Crit Care Med 1999;160:1079-100.
Remmelts HH, Meijvis SC, Biesma DH, van Velzen-Blad H, Voorn GP, Grutters JC, et al
. Dexamethasone downregulates the systemic cytokine response in patients with community-acquired pneumonia. Clin Vaccine Immunol 2012;19:1532-8.
Thatcher TH, Hsiao HM, Pinner E, Laudon M, Pollock SJ, Sime PJ, et al
. Neu-164 and Neu-107, two novel antioxidant and anti-myeloperoxidase compounds, inhibit acute cigarette smoke-induced lung inflammation. Am J Physiol Lung Cell Mol Physiol 2013;305:L165-74.
Ge LT, Liu YN, Lin XX, Shen HJ, Jia YL, Dong XW, et al
. Inhalation of ambroxol inhibits cigarette smoke-induced acute lung injury in a mouse model by inhibiting the Erk pathway. Int Immunopharmacol 2016;33:90-8.
Lee H, Yu SR, Lim D, Lee H, Jin EY, Jang YP, et al
. Galla Chinensis
attenuates cigarette smoke-associated lung injury by inhibiting recruitment of inflammatory cells into the lung. Basic Clin Pharmacol Toxicol 2015;116:222-8.
Hong MH, Kim JH, Na SH, Bae H, Shin YC, Kim SH, et al
. Inhibitory effects of Paeonia suffruticosa
on allergic reactions by inhibiting the NF-kappaB/I kappaB-alpha signaling pathway and phosphorylation of ERK in an animal model and human mast cells. Biosci Biotechnol Biochem 2010;74:1152-6.
Yang XZ, Zhu JY, Tang CP, Ke CQ, Lin G, Cheng TY, et al
. Alkaloids from roots of Stemona sessilifolia
and their antitussive activities. Planta Med 2009;75:174-7.
Liu CL, Cheng L, Ko CH, Wong CW, Cheng WH, Cheung DW, et al
. Bioassay-guided isolation of anti-inflammatory components from the root of Rehmannia glutinosa
and its underlying mechanism via inhibition of iNOS pathway. J Ethnopharmacol 2012;143:867-75.
Li L, Bao H, Wu J, Duan X, Liu B, Sun J, et al
. Baicalin is anti-inflammatory in cigarette smoke-induced inflammatory models in vivo
and in vitro
: A possible role for HDAC2 activity. Int Immunopharmacol 2012;13:15-22.
Lixuan Z, Jingcheng D, Wenqin Y, Jianhua H, Baojun L, Xiaotao F. Baicalin attenuates inflammation by inhibiting NF-kappaB activation in cigarette smoke induced inflammatory models. Pulm Pharmacol Ther 2010;23:411-9.
Gardi C, Stringa B, Martorana PA. Animal models for anti-emphysema drug discovery. Expert Opin Drug Discov 2015;10:399-410.
Yan H, Zhao L, Wu X, Liu H, Wu C, Li Y, et al
. Inflammation and pathological damage to the lungs of mice are only partially reversed following smoking cessation on subacute exposure to cigarette smoke. Mol Med Rep 2015;11:4246-54.
Kubo S, Kobayashi M, Masunaga Y, Ishii H, Hirano Y, Takahashi K, et al
. Cytokine and chemokine expression in cigarette smoke-induced lung injury in guinea pigs. Eur Respir J 2005;26:993-1001.
Shapiro SD. The macrophage in chronic obstructive pulmonary disease. Am J Respir Crit Care Med 1999;160:S29-32.
Barnes PJ. Alveolar macrophages in chronic obstructive pulmonary disease (COPD). Cell Mol Biol (Noisy-le-grand) 2004;50:OL627-37.
Larsson K. Aspects on pathophysiological mechanisms in COPD. J Intern Med 2007;262:311-40.
Meijer M, Rijkers GT, van Overveld FJ. Neutrophils and emerging targets for treatment in chronic obstructive pulmonary disease. Expert Rev Clin Immunol 2013;9:1055-68.
Barnes PJ, Shapiro SD, Pauwels RA. Chronic obstructive pulmonary disease: Molecular and cellular mechanisms. Eur Respir J 2003;22:672-88.
Singh D, Edwards L, Tal-Singer R, Rennard S. Sputum neutrophils as a biomarker in COPD: Findings from the ECLIPSE study. Respir Res 2010;11:77.
Stănescu D, Sanna A, Veriter C, Kostianev S, Calcagni PG, Fabbri LM, et al
. Airways obstruction, chronic expectoration, and rapid decline of FEV1 in smokers are associated with increased levels of sputum neutrophils. Thorax 1996;51:267-71.
Caramori G, Pandit A, Papi A. Is there a difference between chronic airway inflammation in chronic severe asthma and chronic obstructive pulmonary disease? Curr Opin Allergy Clin Immunol 2005;5:77-83.
Tanni SE, Pelegrino NR, Angeleli AY, Correa C, Godoy I. Smoking status and tumor necrosis factor-alpha mediated systemic inflammation in COPD patients. J Inflamm (Lond) 2010;7:29.
Ito K, Lim S, Caramori G, Chung KF, Barnes PJ, Adcock IM. Cigarette smoking reduces histone deacetylase 2 expression, enhances cytokine expression, and inhibits glucocorticoid actions in alveolar macrophages. FASEB J 2001;15:1110-2.
Thomas PS, Yates DH, Barnes PJ. Tumor necrosis factor-alpha increases airway responsiveness and sputum neutrophilia in normal human subjects. Am J Respir Crit Care Med 1995;152:76-80.
Barnes PJ. Cytokine-directed therapies for the treatment of chronic airway diseases. Cytokine Growth Factor Rev 2003;14:511-22.
Li YT, He B, Wang YZ. Exposure to cigarette smoke upregulates AP-1 activity and induces TNF-alpha overexpression in mouse lungs. Inhal Toxicol 2009;21:641-7.
Lee H, Jung KH, Park S, Kil YS, Chung EY, Jang YP, et al
. Inhibitory effects of Stemona tuberosa
on lung inflammation in a subacute cigarette smoke-induced mouse model. BMC Complement Altern Med 2014;14:513.
Keatings VM, Collins PD, Scott DM, Barnes PJ. Differences in interleukin-8 and tumor necrosis factor-alpha in induced sputum from patients with chronic obstructive pulmonary disease or asthma. Am J Respir Crit Care Med 1996;153:530-4.
Botelho FM, Nikota JK, Bauer CM, Morissette MC, Iwakura Y, Kolbeck R, et al
. Cigarette smoke-induced accumulation of lung dendritic cells is interleukin-1α-dependent in mice. Respir Res 2012;13:81.
Zeidel A, Beilin B, Yardeni I, Mayburd E, Smirnov G, Bessler H. Immune response in asymptomatic smokers. Acta Anaesthesiol Scand 2002;46:959-64.
Chung KF. Cytokines as targets in chronic obstructive pulmonary disease. Curr Drug Targets 2006;7:675-81.
Ekberg-Jansson A, Andersson B, Bake B, Boijsen M, Enanden I, Rosengren A, et al
. Neutrophil-associated activation markers in healthy smokers relates to a fall in DL (CO) and to emphysematous changes on high resolution CT. Respir Med 2001;95:363-73.
Hammad DR, Elgazzar AG, Essawy TS, Abd El Sameie SA. Evaluation of serum interleukin-1 beta as an inflammatory marker in COPD patients. Egypt J Chest Dis Tuberc 2015;64:347-52.
Yao X, Huang J, Zhong H, Shen N, Faggioni R, Fung M, et al
. Targeting interleukin-6 in inflammatory autoimmune diseases and cancers. Pharmacol Ther 2014;141:125-39.
Song W, Zhao J, Li Z. Interleukin-6 in bronchoalveolar lavage fluid from patients with COPD. Chin Med J (Engl) 2001;114:1140-2.
Agustí A, Edwards LD, Rennard SI, MacNee W, Tal-Singer R, Miller BE, et al
. Persistent systemic inflammation is associated with poor clinical outcomes in COPD: A novel phenotype. PLoS One 2012;7:e37483.
Celli BR, Locantore N, Yates J, Tal-Singer R, Miller BE, Bakke P, et al
. Inflammatory biomarkers improve clinical prediction of mortality in chronic obstructive pulmonary disease. Am J Respir Crit Care Med 2012;185:1065-72.
de Boer WI, Sont JK, van Schadewijk A, Stolk J, van Krieken JH, Hiemstra PS. Monocyte chemoattractant protein 1, interleukin 8, and chronic airways inflammation in COPD. J Pathol 2000;190:619-26.
Traves SL, Culpitt SV, Russell RE, Barnes PJ, Donnelly LE. Increased levels of the chemokines GROalpha and MCP-1 in sputum samples from patients with COPD. Thorax 2002;57:590-5.
Macnee W. Pathogenesis of chronic obstructive pulmonary disease. Clin Chest Med 2007;28:479-513, v.
Barnes PJ. The cytokine network in chronic obstructive pulmonary disease. Am J Respir Cell Mol Biol 2009;41:631-8.
Dinarello CA. Proinflammatory cytokines. Chest 2000;118:503-8.
Lim S, Roche N, Oliver BG, Mattos W, Barnes PJ, Chung KF. Balance of matrix metalloprotease-9 and tissue inhibitor of metalloprotease-1 from alveolar macrophages in cigarette smokers. Regulation by interleukin-10. Am J Respir Crit Care Med 2000;162:1355-60.
Churg A, Zhou S, Wang X, Wang R, Wright JL. The role of interleukin-1ß in murine cigarette smoke–induced emphysema and small airway remodeling. Am J Respir Cell Mol Biol 2009;40:482-90.
Anthony D, McQualter JL, Bishara M, Lim EX, Yatmaz S, Seow HJ, et al
. SAA drives proinflammatory heterotypic macrophage differentiation in the lung via CSF-1R-dependent signaling. FASEB J 2014;28:3867-77.
Aldonyte R, Jansson L, Piitulainen E, Janciauskiene S. Circulating monocytes from healthy individuals and COPD patients. Respir Res 2003;4:11.
Caramori G, Adcock IM, Di Stefano A, Chung KF. Cytokine inhibition in the treatment of COPD. Int J Chron Obstruct Pulmon Dis 2014;9:397-412.
[Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5]