Effect of APN on collagen type III expression in control or PQ-incubated lung fibroblast.

<p>PQ (0.3 mmol/L, 24 hours) increased, whereas pretreated with APN (5 ug/mL, 2 hours) rescued, collagen type III expression in lung fibroblasts at 24 and 72 h after PQ incubation. The collagen type III protein and mRNA expression were a determined via Western blot and RT-PCR, respectively. Values represent the mean ± standard error of the mean of three parallel measurements. * P<0.05. APN, adiponectin; PQ, paraquat.</p

Effect of APN on TGF-β1, CTGF and α-SMA expression in lung tissues from control animals or PQ-exposed animals, treated with vehicle or APN. PQ (10 mg/kg, IP) increased, whereas APN (100 μg/kg or 500μg/kg, IV) revised, TGF-β1 (A), CTGF (B) and α-SMA (C) expression in mice lung tissues at 3, 14 and 21 d after PQ injection.

<p>The protein and mRNA expression were a determined via Western blot and RT-PCR, respectively. Values represent the mean ± standard error of the mean of four parallel measurements. *P<0.05. APN, adiponectin; TGF-β1, transforming growth factor-β1; CTGF, connective tissue growth factor; α-SMA, α-smooth muscle actin; PQ, paraquat.</p

Lung fibrosis scores across groups.

<p>*: PQ group VS control group, <i>p<0</i>.<i>05</i></p><p>^#:PQ + APN group VS PQ group, <i>p<0</i>.<i>05</i></p><p>Lung fibrosis scores across groups.</p

APN receptor expression in lung fibroblast.

<p>Both AdipoR1 and AdipoR2 expression were detected in WI-38 lung fibroblasts. PQ (0.3 mmol/L, 24 hours) treatments up-regulated protein and mRNA expression of both APN receptors in lung fibroblasts WI-38 at 24, 48 and 72 h after PQ incubation. APN (5 ug/mL, 2 hours) pretreatment further enhanced AdipoR1, but not AdipoR2, up-regulation. AdipoR1-specific siRNA successfully abrogated AdipoR1 expression. AdipoR1 and AdipoR2 expression were determined by Western blot and RT-PCR. Values represent the mean ± standard error of the mean of four parallel measurements. *P<0.05. APN, adiponectin; PQ, paraquat, AdipoR, APN receptor.</p

AdipoR1-specific siRNA reversed the protective effect of APN against PQ-mediated fibrosis in lung fibroblasts WI-38 at 72 h after PQ incubation.

<p>Cell viability, apoptosis and collagen type III mRNA expression in lung fibroblasts were measured via CCK-8 assay, Annexin V-FITC/PI staining with flow cytometry, and RT-PCR, respectively. Values represent the mean ± standard error of the mean of three parallel measurements. *P<0.05. APN, adiponectin; PQ, paraquat, AdipoR, APN receptor.</p

Effect of APN on TIMP-1 blood levels in control animals or PQ-exposed animals, treated with vehicle or APN.

<p>PQ (10 mg/kg, IP) increased, whereas APN (100 μg/kg or 500μg/kg, IV) reversed, TIMP-1 levels at 3, 7, 14and 21 d after PQ injection. TIMP-1 levels were determined by an enzyme-linked immunosorbent assay. Values represent the mean ± standard error of the mean of four parallel measurements. *P<0.05. APN, adiponectin; TIMP-1, tissue inhibitor of metalloproteinase-1; PQ, paraquat.</p

Histological evaluation of mice lungs in the control, PQ, PQ+ low-dose APN or PQ+ high-dose APN group on day 3, 7, 14, 21 after PQ injection with Masson stain (×200 magnification).

<p>Compared to control, PQ (10 mg/kg) exposure increased fibrosis (seen as blue collagen deposition) in the alveolar regions and small bronchioles from day 3, peaking on day 21. Both low-dose (100 μg/kg) and high-dose (500 μg/kg) APN groups exhibited decreased collagen deposition, in a dose-dependent manner.</p

Primer sequences used for RT-PCR.

<p>Primer sequences used for RT-PCR.</p

Correlation between DR and CTRPs.

<p>Correlation between DR and CTRPs.</p

Correlation between DR and laboratory characteristics.

<p>Correlation between DR and laboratory characteristics.</p