Herbal Formula, PM014, Attenuates Lung Inflammation in a Murine Model of Chronic Obstructive Pulmonary Disease

Abstract

Chronic obstructive pulmonary disease (COPD), which is characterized by airway obstruction, leads to, as the two major forms of COPD, chronic bronchitis and emphysema. This study was conducted to evaluate the effects of herbal formula, PM014, in a murine model of COPD. Balb/c mice were treated once with each herb extract in PM014 or PM014 mixture via an oral injection. Lipopolysaccharide (LPS) or elastase/LPS were administrated to the mice to induce a disease that resembles COPD. PM014 treatment significantly attenuated the increased accumulation of immune cells in bronchoalveolar lavage fluid (BALF) compared to control mice. In addition, the TNF-α and IL-6 levels in BALF were decreased in the PM014 mice. Furthermore, histological analysis demonstrated that PM014 attenuated the hazardous effects of lung inflammation. These data suggest that PM014 exerts beneficial effects against forms of COPD such as lung inflammation.

Figure 1

A representative DART-MS spectrum of PM104 extract.

Figure 2

The therapeutic effect of mixed herbs, PM014, and individual herbs. Lipopolysaccaride was administrated to mice to induce a disease that resembles COPD. The numbers of neutrophils, macrophages, lymphocytes, and total cells were determined in BAL fluid. Con: control, LPS: lipopolysaccaride, DXT: dexametasone, RG: Rehmannia glutinosa, PS: Paeonia suffruticosa, SC: Schizandra chinensis, AC: Asparagus cochinchinensis, PA: Prunus armeniaca, SB: Scutellaria baicalensis, SS: Stemona sessilifolia. Data are expressed as the mean number of cells ± S.E.M. (*P < 0.05, **P < 0.01, ***P < 0.001 versus LPS; n = 4–6).

Figure 3

Effect of PM014 on immune cell profiles and pro-inflammatory cytokines. (a) Effect of PM014 on immune cell profiles in BAL fluid. The numbers of neutrophils, macrophages, lymphocytes, and total cells were determined in BAL fluid. (b) The levels of IL-6 and TNF-α in BAL fluid were determined by ELISA. Data are expressed as the mean number of cells ± SEM (*P < 0.05, **P < 0.01, ***P < 0.001 versus LPS; n = 6).

Figure 4

Effect of PM014 on lung tissue. LPS treatment induced parenchymal and airway inflammation. Mouse lung sections were stained with hematoxylin and eosin (magnification ×200).

Figure 5

Effect of PM014 on the expression of neutrophil elastase and proliferating cell nuclear antigen. Mouse lung sections were stained immunohistochemically. Mouse lung sections were stained with anti-NE goat polyclonal antibody (a) or with anti-PCNA goat polyclonal antibody (b). After the slides were incubated with avidin-biotin peroxidase complex, the color was developed with 3,3′-diaminobenzidine tetrachloride.

Figure 6

Effect of PM014 on lung inflammation in elastase/LPS-exposed mice. (a) Experimental plan of repeated elastase and LPS exposure. (b) Effect of PM014 on immune cell profiles in BAL fluid. The numbers of neutrophils, macrophages, lymphocytes, and total cells were determined in BAL fluid. (c) The levels of IL-6 and TNF-α in BAL fluid were determined by ELISA. PPE: porcine pancreatic elastase, LPS: lipopolysaccharide. Data are expressed as the mean number of cells ± SEM (*P < 0.05, **P < 0.01 versus LPS; n = 5-6).

Figure 7

Effect of PM014 on lung tissue changes in elastase/LPS-exposed mice. (a) Mouse lung sections were stained with hematoxylin and eosin (magnification ×200). (b) Mouse lung sections were stained with PAS (magnification ×400). PPE: porcine pancreatic elastase, LPS: lipopolysaccaride.