Inhaled corticosteroid normalizes some but not all airway vascular remodeling in COPD

Figures & data

Figure 1 Rbm and LP vessels stained with anti-Collagen IV antibody in (A) and anti-Factor VIII antibody in (B). The epithelium sits on the basement membrane. The thickness of the Rbm, is shown with the two-headed arrow. Vessels are in contact or embedded within the Rbm (arrows). Arrowheads indicate vessels in the lamina propria. Magnification ×400; scale bar =20 µm.

Abbreviations: Rbm, reticular basement membrane; LP, lamina propria.

Figure 2 Study subjects and design.

Abbreviations: FP, fluticasone propionate; COPD, chronic obstructive pulmonary disease.

Table 1 Demographics of the study groupsa

Groups (numbers)	ICS (n=23)	Placebo (n=11)	Normal nonsmoking control (n=8)
Age, years	61 (46–69)	61 (52–69)	54 (32–68)
Female/male	9/14	4/7	2/6
Current smoker/ex-smoker	13/10	4/7	NA
Pack-year smoking history	44 (18–150)	51 (22–148)	0
GOLD stage I/II	12/11	5/6	NA
FEV1/FVC ratio	59 (41–68)	57 (38–68)	79 (71–88)b
%DLCO predicted	65 (44–87)	66 (45–90)	–
TLC, L	7.6 (5.3–8.4)	6.2 (5.1–9.8)	–
RV, L	2.8 (2.1–4.0)	2.4 (1.6–4.7)	–

Notes: Data from a normal control group have been added to this table from a comparable previous study. Adapted from Soltani A, Wood-Baker R, Sohal SS, Muller HK, Reid D, Walters EH. Reticular basement membrane vessels are increased in COPD bronchial mucosa by both factor VIII and collagen IV immunostaining and are hyperpermeable. J Allergy (Cairo). 2012:958383. Creative Commons license and disclaimer available from: http://creativecommons.org/licenses/by/4.0/legalcode.⁷ This additional material was used for the comparison made in Figure 3 between normal controls and both treatment groups before and after interventions.

a All data in the table are presented as median (range);

b significantly different between normal controls and COPD groups. “–”, no data.

Abbreviations: DLCO, diffusion capacity of lung diffusion for carbon monoxide; FEV₁, forced expiratory volume in 1 second; FVC, forced vital capacity; GOLD, Global initiative for chronic Obstructive Lung Disease; ICS, inhaled corticosteroid; NA, not applicable; RV, residual volume; TLC, total lung capacity.

Table 2 Comparison of baseline tissue vascular parameters between groupsa

Groupsb	ICS	Placebo	P-values
No of Rbm vessels/mm Rbm	5.1 (0.0–15.2)	3.6 (1.8–6.5)	0.5
No of vessels in the Rbm stained for VEGF/mm Rbm	0.6 (0.0–5.4)	1.0 (0.0–5.4)	0.9
No of vessels stained for TGF-β1/mm Rbm	1.3 (0.0–8.1)	2.5 (0.7–4.0)	1.0
Density of LP vessels number/mm^2	289 (158–585)	277 (200–641)	0.7
No of vessels stained for VEGF in the LP/mm^2	113 (21–276)	144 (16–366)	0.5

Notes: Also see Figure 1 and Figures S1 and S2.

a All data in the table are presented as median (range);

b numbers of subjects for ICS vs placebo for Factor VIII vessel staining were 13 vs 6; for VEGF staining 16 vs 7; and for TGF-β1 staining 8 vs 3.

Abbreviations: ICS, inhaled corticosteroid; LP, lamina propria; Rbm, reticular basement membrane.

Figure 3 The effects of ICS or placebo on LP vessels. The box plot of H-N shows that the LP was hypovascular in both treatment groups. Bars indicate medians. Dots and triangles represent current smoking and ex-smoking COPD subjects. There was strong trend for an increase in vessels overall, but this was confined to active smokers in whom the change was significant (P=0.05).

Abbreviations: LP, lamina propria; H-N, healthy nonsmokers; ICS, inhaled corticosteroid; COPD, chronic obstructive pulmonary disease.

Table 3 Changes with treatmenta

Measurements	ICS			Placebo
	Before	After	P-values	Before	After	P-values
Number of Rbm vessels/mm Rbm	5.1 (0.0–15.2)	2.3 (0.8–13)	0.5	3.6 (1.8–6.5)	2.8 (1.3–10.8)	0.9
Number of LP vessels/mm^2 of LP	289 (158–585)	386 (213–444)	0.08	277 (200–641)	295 (173–377)	0.5
% of vessels stained for VEGF/mm Rbmc	19% (0–256)	16% (0–302)	0.5	0% (0–49)	19% (0–250)	0.9
% of vessels stained for TGF-β1/mm Rbmb,c	47% (0–154)	94% (0–1,416)	0.3	–	–	–
% of vessels stained for p-Smad 2/3/mm Rbmb,c	69% (46–119)	0% (0–89)	0.1	–	–	–
% of vessels stained for VEGF/mm^2 of LPc	61% (7–90)	35% (0–130)	0.2	32% (5–180)	88% (26–256)	0.3
% of vessels stained for p-Smad 2/3/mm^2 of LPb,c	24% (4–80)	10% (5–43)	0.03	–	–	–
TLC	7.6 (5.3–8.4)	7.6 (5.2–8.2)	0.4	6.2 (5.1–9.8)	6.3 (5.8–9.4)	0.5
RV	2.8 (2.1–4.0)	2.8 (2.0–4.2)	0.7	2.4 (1.6–4.7)	2.6 (2.1–4.5)	0.4

Notes:

a All data in the table are presented as median (range);

b some cells do not contain numbers because the number of paired tissues was too small for formal analysis (the number of paired tissues for TGF-β1 and SMAD 2/3 in the placebo arm was three;);

c percentage of vessels stained for VEGF/millimeter Rbm, percentage of vessels stained for TGF-β1/mm Rbm, percentage of vessels stained for p-SMAD2/3/mm Rbm, percentage of vessels stained for VEGF/mm² of LP, and percentage of vessels stained for p-SMAD 2/3/mm² of LP are calculated as vessels stained for VEGF, TGF-β1, p-SMAD 2/3×100/total number of vessels stained for Factor VIII per mm of the Rbm or per mm² of LP, respectively. Also see Figure 1 and Figures S1 and S2. “–”, no data.

Abbreviations: ICS, inhaled corticosteroid; LP, lamina propria; Rbm, reticular basement membrane; TLC, total lung capacity; RV, residual volume.

Figure 4 Significant correlation between the baseline number and change in LP vessels with ICS (fluticasone propionate) (r=−0.7, P=0.01). Circles and triangles present current smoking and ex-smoking COPD subjects, respectively.

Abbreviations: LP, lamina propria; ICS, inhaled corticosteroid; COPD, chronic obstructive pulmonary disease.

Figure S1 VEGF-stained vessels in the Rbm and LP are pointed out with narrow arrows and wide arrows, respectively.

Notes: The lamina propria is situated beneath the Rbm. The width of the Rbm is shown by a two-headed arrow. Magnification ×400.

Abbreviations: Rbm, reticular basement membrane; LP, lamina propria.

Figure S2 TGF-β-stained vessels in the Rbm are pointed out with arrows. The generalized dark immunostaining in the lamina propria, which is situated beneath the Rbm, impedes vessel identification. The width of the Rbm is shown by a two-headed arrow. Magnification ×400.

Abbreviation: Rbm, reticular basement membrane.

SoltaniAWood-BakerRSohalSSMullerHKReidDWaltersEHReticular basement membrane vessels are increased in COPD bronchial mucosa by both factor VIII and collagen IV immunostaining and are hyperpermeableJ Allergy (Cairo)201295838322500190

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