Over-expression of nuclear factor-κB family genes and inflammatory molecules is related to chronic obstructive pulmonary disease

Figures & data

Table 1 Primer sequences of candidate genes

Gene name	Primer	Sequence
β-actin	Forward	5′-CTGGAACGGTGAAGGTGACA-3′
	Reverse	5′-CGGCCACATTGTGAACTTTG-3′
NF-κB1	Forward	5′-CACAAGGCAGCAAATAGACGAG-3′
	Reverse	5′-TGGGGCATTTTGTTGAGAGTT-3′
NF-κB2	Forward	5′-GGCTGGTGCTGACATCCAT-3′
	Reverse	5′-CTGCTTCGGGTGTCCTTCTC-3′
c-REL	Forward	5′-GACGACTGCTCTTCCTCCTGTT-3′
	Reverse	5′-TCATCTCCTCCTCTGACACTTCC-3′

Table 2 Baseline characteristics of the COPD group and control group

	COPD group (n = 186)				Control group (n = 180) (%)	t/χ^2	P-value
	Moderate (n = 53) (%)	Severe (n = 60) (%)	Very severe (n = 73) (%)	Total (%)
Age	62.21 ± 13.22	63.25 ± 10.92	59.43 ± 9.62	63.17 ± 11.54	63.03 ± 11.53	0.115	0.909
Sex
Male	24 (45.3)	30 (50.0)	43 (58.9)	97 (52.2%)	77 (42.8)	3.222	0.073
Female	29 (54.7)	30 (50.0)	30 (41.1)	89 (47.8%)	103 (57.2)
Smoking
No	35 (66.0)	36 (60.0)	35 (47.9)	106 (53.0%)	135 (75.0)	13.195	<0.001
Yes	18 (34.0)	24 (40.0)	38 (52.1)	80 (47.0%)	45 (25.0)
Pack years	45.00 ± 44.75	40.00 ± 27.50	20.00 ± 30.00

Notes: Age and pack years are expressed as mean ± standard error of the mean. The difference in age between the COPD group and the control group was compared with Student’s t-test. Sex and smoking are expressed as n (%). The differences were compared with Pearson’s chi-square test.

Abbreviation: COPD, chronic obstructive pulmonary disease.

Figure 1 Relative mRNA levels of NF-κB family genes in COPD group and control group. Relative mRNA levels of NF-κB family genes were expressed as the mean value ± standard deviation. *P < 0.05, compared with the control group.

Abbreviations: NF-κB, nuclear factor-κB; COPD, chronic obstructive pulmonary disease.

Figure 2 Relative mRNA levels of NF-κB family genes among patients with different severities of COPD. Relative mRNA levels of NF-κB family genes were expressed as the mean value ± standard deviation.

Abbreviations: NF-κB, nuclear factor-κB; COPD, chronic obstructive pulmonary disease.

Table 3 Differences in inflammatory molecules levels between COPD group and control group (mean ± standard deviation)

Inflammatory molecules	COPD group (n = 186)	Control group (n = 180)	t	P-value	SD (95% CI)a
COX-2 (U/L)	71.60 ± 9.40	50.64 ± 0.09	21.508	<0.001	1.328 (1.249–1.411)
CRP (mg/L)	6.20 ± 1.21	4.15 ± 1.03	17.256	<0.001	6.088 (4.162–0.904)
IL-1β (pg/mL)	43.33 ± 5.70	28.60 ± 5.07	25.894	<0.001	2.334 (1.845–2.951)
IL-6 (pg/mL)	29.90 ± 5.32	20.57 ± 3.37	19.858	<0.010	1.818 (1.590–2.079)
IL-8 (pg/mL)	76.28 ± 13.29	49.99 ± 10.71	20.661	<0.001	1.221 (1.169–1.274)
IL-12 (pg/mL)	30.42 ± 5.72	21.74 ± 4.05	16.609	<0.001	1.495 (1.372–1.628)
TNF-α (pg/mL)	45.35 ± 8.25	29.24 ± 4.25	23.287	<0.001	1.530 (1.388–1.687)

Note:

a Compared with controls after adjustment for smoking.

Abbreviations: COPD, chronic obstructive pulmonary disease; SD, standard deviation; CI, confidence interval; COX-2, cyclooxygenase-2; CRP, C-reactive protein; IL, interleukin; TNF-α, tumor necrosis factor-α.

Table 4 Differences in inflammatory molecules levels between smokers and nonsmokers with or without COPD

Inflammatory molecules	Control group		COPD group		F	P-value
	Nonsmoker	Smoker	Nonsmoker	Smoker
COX-2 (U/L)	51.09 ± 8.74	49.30 ± 10.06	71.20 ± 9.56^a,b	72.14 ± 9.21^a,b	154.662	<0.001
CRP (mg/L)	4.13 ± 1.05	4.21 ± 0.96	6.09 ± 1.16^a,b	6.34 ± 1.26^a,b	100.269	<0.001
IL-1β (pg/mL)	28.48 ± 5.05	28.97 ± 5.17	42.90 ± 5.53^a,b	43.90 ± 5.91^a,b	223.974	<0.001
IL-6 (pg/mL)	20.55 ± 3.37	20.63 ± 3.44	29.51 ± 4.97^a,b	30.42 ± 5.75^a,b	131.998	<0.010
IL-8 (pg/mL)	49.90 ± 10.73	50.28 ± 10.76	77.75 ± 13.10^a,b	74.31 ± 13.37^a,b	144.158	<0.001
IL-12 (pg/mL)	21.82 ± 3.94	21.51 ± 4.41	29.75 ± 5.24^a,b	31.31 ± 6.23^a,b	94.101	<0.001
TNF-α (pg/mL)	29.19 ± 4.23	29.38 ± 4.35	44.68 ± 7.81^a,b	46.23 ± 8.78^a,b	181.852	<0.001

Notes:

a P < 0.05, compared with nonsmoker controls.

b P < 0.05, compared with smoker controls.

Abbreviations: COPD, chronic obstructive pulmonary disease; COX-2, cyclooxygenase-2; CRP, C-reactive protein; IL, interleukin; TNF-α, tumor necrosis factor-α.

Table 5 Differences in inflammatory molecules levels among patients with different severities of COPD

Inflammatory molecules	Different severities of COPD			F	P-value
	Moderate (n = 53)	Severe (n = 60)	Very severe (n = 73)
COX-2 (U/L)	70.98 ± 10.13	71.49 ± 9.41	72.64 ± 8.75	0.374	0.689
CRP (mg/L)	6.07 ± 1.16	6.36 ± 0.94	6.22 ± 1.44	0.583	0.560
IL-1β (pg/mL)	43.89 ± 5.45	42.61 ± 5.63	43.33 ± 5.44	0.537	0.586
IL-6 (pg/mL)	28.48 ± 4.90	30.53 ± 4.97	30.53 ± 6.46	1.753	0.177
IL-8 (pg/mL)	74.77 ± 13.79	76.90 ± 12.91	74.97 ± 12.86	0.336	0.715
IL-12 (pg/mL)	28.76 ± 5.03	30.85 ± 5.27	31.03 ± 6.95	1.768	0.175
TNF-α (pg/mL)	43.78 ± 8.00	46.97 ± 7.46	45.65 ± 9.70	1.350	0.263

Abbreviations: COPD, chronic obstructive pulmonary disease; COX-2, cyclooxygenase-2; CRP, C-reactive protein; IL, interleukin; TNF-α, tumor necrosis factor-α.

Figure 3 Subgroup analysis of inflammatory molecules levels based on the mRNA expression levels of NF-κB family genes in patients with COPD: (A) IL-1β, (B) IL-6, (C) IL-8, (D) IL-12, (E) TNF-α, (F) COX-2 and (G) CRP. *P < 0.05, compared with over-expression subgroup.

Abbreviations: NF-κB, nuclear factor-κB; COPD, chronic obstructive pulmonary disease; IL, interleukin; TNF-α, tumor necrosis factor-α; COX-2, cyclooxygenase-2; CRP, C-reactive protein.