Aerobic Exercise Alleviates Inflammation, Oxidative Stress, and Apoptosis in Mice with Chronic Obstructive Pulmonary Disease

Figures & data

Table 1 PCR Primer Sequence Information of Mouse

Gene Name	Primer Sequence
IL-1β	F: 5ʹ GCTCATTGCTGGGTACTTACAA 3ʹ
	R: 5ʹCCAGACTTGGCACAAGACAGG 3ʹ
1L-10	F: 5ʹ GCTCTTACTGACTGGCATGAG 3ʹ
	R: 5ʹ CGCAGCTCTAGGAGCATGTG 3ʹ
IL-17	F: 5ʹ CCAAGAGGTGAGTGCTTCCC 3ʹ
	R: 5ʹ CTGTTGTTCAGACTCTCTCCCT 3ʹ
TNF-α	F: 5ʹ ATGTCTCAGCCTCTTCTCATTC 3ʹ
	R: 5ʹ GCTTGTCACTCGAATTTTGAGA 3ʹ
CXCL1	F: 5ʹ CTGGGATTCACCTCAAGAACATC 3ʹ
	R: 5ʹCAGGGTCAAGGCAAGCCTC 3ʹ
Sirt1	F: 5ʹ GCTGACGACTTCGACGACG 3ʹ
	R: 5ʹ TCGGTCAACAGGAGGTTGTCT 3ʹ
MMP-9	F: 5ʹ TCTTGGAGTAAGTCGAGAAGTGT 3ʹ
	R: 5ʹ GTTGAAACTGAGCGAAAAAGGC 3ʹ
GAPDH	F: 5ʹ AGGTCGGTGTGAACGGATTTG 3ʹ
	R: 5ʹ TGTAGACCATGTAGTTGAGGTCA 3ʹ

Abbreviation: PCR, polymerase chain reaction.

Table 2 Total and Differential Cell Counts in BALF (Cells/ml)

	Con	COPD	COPD + AE	P
Total cells	1.29 ± 0.86	9.98 ± 2.14^#	8.08 ± 0.94*	< 0.001
Neutrophils	0.04 ± 0.04	7.55 ± 1.23^#	5.84 ± 1.38*	< 0.001
Lymphocytes	0.03 ± 0.05	0.05 ± 0.14	0.05 ± 0.16	0.88
Macrophages	1.09 ± 0.48	2.12 ± 0.77	1.26 ± 0.57	0.007
Eosinophils	0.01± 0.03	0.01 ± 0.03	0.04 ± 0.05	0.21

Notes: Compared with the Con group, LPS administration increased the number of total cells, neutrophils, and macrophages. Compared with the LPS group, exercise administration decreased the number of total cells, neutrophils, and macrophages. ^#Indicates that the difference in the data is significant compared with the Con group *Indicates that the difference in the data is significant compared with the COPD group. Values are expressed as the means ± SD.

Table 3 The Levels of Inflammatory Factors in BALF and Serum (pg/ml)

	Con	COPD	COPD + AE	P
CXCL1 BALF	9.24 ± 3.65	1106.6 ± 357.31^#	1648.91 ± 256.32*	< 0.001
IL-17 BALF	8.31 ± 1.33	254.16 ± 97.69^#	251.44 ± 108.92	< 0.001
TGF-β BALF	21.35 ± 4.86	165.68 ± 63.75^#	98.45 ± 44.73*	< 0.001
IL-10 BALF	26.9 ± 20.7	503.2 ± 95.85^#	834.9 ± 216.7*	< 0.001
IL-1β BALF	12.65 ± 2.14	706.9 ± 141.5^#	563.1 ± 106.93*	< 0.001
TNF-α BALF	13.31 ± 5.92	152.3 ± 28.9^#	124.6 ± 13.9*	< 0.001
IL-10 serum	13.19 ± 15.65	506.6 ± 157.31^#	748.9 ± 143.3*	< 0.001
TNF-α serum	14.24 ± 8.46	54.45 ± 16.81^#	31.1 ± 10.21*	<0.001

Notes: The levels of CXCL1, IL-17, TGF-β, IL-10, IL-1β, and TNF-α in BALF and the levels of IL-10 and TNF-α in serum were detected. ^#Indicates that the difference in the data is significant compared with the Con group, *Indicates that the difference in the data is significant compared with the COPD group. Values are expressed as the means ± SD.

Abbreviations: IL, interleukin; TNF, tumor necrosis factor; CXCL, C-X-C motif chemokine ligand; TGF, transforming growth factor.

Figure 1 Detection of emphysema.

Notes: (A) Con group. (B) COPD group. (C) COPD+AE group. Hematoxylin-eosin staining revealed evidence of the degree of emphysema. (D) Detection of MLI. LPS administration increased the MLI, while 4 weeks of exercise intervention significantly decreased the MLI. The difference in MLI was significant between the Con and COPD + AE groups. ^#P<0.05 compared with the Con group. *P<0.05 compared with the COPD group (magnification, x100).

Abbreviation: MLI, mean linear intercept.

Figure 2 Detection of pulmonary fibrosis.

Notes: (A) Con group. (B) COPD group. (C) COPD+AE group. (D) Detection of the fibrotic score. Mason staining revealed evidence of the degree of pulmonary fibrosis. The fibrotic score increased after LPS administration, while exercise administration decreased fibrotic score. ^#P<0.05 compared with the Con group. *P<0.05 compared with the COPD group (magnification, x200).

Figure 3 Detection of airway fibrosis.

Notes: (A) Con group. (B) COPD group. (C) COPD+AE group. (D) Detection of collagen fiber deposition in the airway wall. The airway collagen increased after LPS administration, while exercise administration decreased fibrotic score. The results were expressed as μm² of collagen fibers per μm² of tissue area. ^#P<0.05 compared with the Con group. *P<0.05 compared with the COPD group (magnification, x40).

Figure 4 Detection of gene expression levels in lung tissue.

Notes: The gene expression levels of pro-inflammatory factors IL-1β (A), IL-17 (C), TNF-α (D), and CXCL1 (E) and profibrotic factors TGF-β (F) and MMP-9 (G) and decreased anti-inflammatory factor IL-10 (B) and lung-protective factor sirt1 (H) were detected. ^#P<0.05 compared with the Con group. *P<0.05 compared with the COPD group.

Figure 5 Detection of cell apoptosis.

Notes: (A) Con group. (B) COPD group. (C) COPD+AE group. (D) Detection of apoptosis index. TUNEL staining revealed evidence of cell apoptosis in the lung. LPS administration increased the number of apoptotic cells (red arrow) and apoptosis index. 4 weeks of exercise intervention decreased the number of apoptotic cells and apoptosis index in COPD mice. ^#P<0.05 compared with the Con group. *P<0.05 compared with the COPD group (magnification, x200).

Figure 6 Detection of oxidative stress injury.

Notes: The levels of MDA (A), MPO (B), GSH (C), and SOD (D) as indicators of oxidative stress injury were detected. LPS administration increased the levels of MDA and MPO and decreased the levels of SOD and GSH. 4 weeks of exercise intervention decreased the levels of MDA and MPO and increased the levels of SOD and GSH in mice with COPD. ^#P<0.05 compared with the Con group. *P<0.05 compared with the COPD group.

Figure 7 Detection of bronchial mucus cell hyperplasia and bronchoconstriction.

Notes: (A) Con group. (B) COPD group. (C) COPD+AE group. LPS administration led to severe bronchial mucus cell hyperplasia and bronchoconstriction, while 4 weeks of AE improved bronchial mucus cell hyperplasia and bronchoconstriction (magnification, x400).