The mechanism underlying alpinetin-mediated alleviation of pancreatitis-associated lung injury through upregulating aquaporin-1

Figures & data

Figure 1 The effect of SAP on lung tissue.

Notes: (A) Twelve hours after establishing the SAP model using ursodeoxycholic acid sodium, the lung tissues from each group were stained by HE for microscopic observation (×200 magnification). (B) The change in the lung W/D. The W/D in the model group was significantly greater compared with the sham group (*P<0.05); each experiment was repeated three times.
Abbreviations: HE, hematoxylin and eosin; SAP, severe acute pancreatitis; W/D, wet-to-dry weight ratio.

Figure 2 The level of AQP-1 expression in the pancreatitis-associated lung injuries measured in the animal models and at the cellular level.

Notes: (A, B) The level of AQP-1 protein expressed in each group was analyzed using Western blot. (C): The level of AQP-1 mRNA expressed in lungs from each group was analyzed using RT-PCR. The results were compared with the control group; *P<0.05. Each experiment was repeated three times.
Abbreviations: AQP-1, aquaporin-1; HPMVEC, human pulmonary microvascular endothelial cell; LPS, lipopolysaccharide; RT-PCR, reverse transcription polymerase chain reaction.

Table 1 The change in the lung W/D (x ± s)

Group	6 hours	12 hours	24 hours
Sham group	2.96±0.35	3.16±0.17	3.34±0.38
Model group	4.12±0.24*	5.51±0.20*	8.08±0.23*
Alpinetin group	3.08±0.20**	3.22±0.19**	3.57±0.21**
AQP-1^−/− group	4.15±0.18	5.62±0.14	8.49±0.20
Dexamethasone group	3.03±0.35**	3.09±0.47**	3.34±0.18**

Notes: Lung W/D measurement. The groups were compared with the sham group, and

* P<0.05. The groups were also compared with the model group, and

** P<0.05. Each experiment was repeated three times.

Abbreviations: AQP-1^−/−, AQP-1 knockout pancreatitis-associated lung injury group; W/D, wet-to-dry weight ratio.

Figure 3 The effect of alpinetin on lung tissue in pancreatitis-associated lung injury.

Notes: (A) The lung tissues from sham group were stained by HE for microscopic observation. (B) The lung tissues from alpinetin group (160 μg/mL) were stained by HE for microscopic observation. (C) Sham group was observed in the AQP-1 expression in lung tissue by immunohistochemistry. (D) Alpinetin group (160 μg/mL) was observed in the AQP-1 expression in lung tissue by immunohistochemistry. The lung tissue was examined using immunohistochemical staining (×100 magnification).
Abbreviations: AQP-1, aquaporin-1; HE, hematoxylin and eosin.

Figure 4 Alpinetin can promote the HPMVEC proliferation.

Notes: Different alpinetin doses (0.01, 0.1, 1.0, and 10.0 μg/mL) were applied to the LPS-HPMVECs for 24, 48, and 72 hours; the MTT assay was used to measure cell viability. Each experiment was repeated three times. *P<0.05.
Abbreviations: HPMVEC, human pulmonary microvascular endothelial cell; LPS, lipopolysaccharide; MTT assay, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoli-umbromide; OD, optical density.

Figure 5 The level of AQP-1 expressed in pancreatitis-associated lung injury was measured in the animal models and at the cellular level.

Notes: (A) The level of AQP-1 protein expressed in the lung tissues of animal experiments was analyzed using Western blot. (B) The level of AQP-1 protein expressed in the lung at the cellular level was analyzed using Western blot. The results were compared with the control group, *P<0.05 and compared with the model group, **P<0.05. Each experiment was repeated three times.
Abbreviations: AQP-1, aquaporin-1; DXM, dextromethorphan; HPMVEC, human pulmonary microvascular endothelial cell; LPS, lipopolysaccharide.

Figure 6 The level of TNF-α expressed in pancreatitis-associated lung injury was measured in the animal models and at the cellular level.

Notes: (A) The level of TNF-α protein expressed for each group in the in vivo experiment was analyzed using Western blot. (B) The level of TNF-α protein expressed for each group at the cellular level was analyzed using Western blot. The results were compared with the control group, *P<0.05 and compared with the model group, **P<0.05. Each experiment was repeated three times.
Abbreviations: AQP-1, aquaporin-1; DXM, dextromethorphan; HPMVEC, human pulmonary microvascular endothelial cell; LPS, lipopolysaccharide; TNF, tumor necrosis factor.