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  • Aldosterone Contributed to Pulmonary Arterial Hypertension Development via Stimulating Aquaporin Expression and Pulmonary Arterial Smooth Muscle Cells Proliferation.

Aldosterone Contributed to Pulmonary Arterial Hypertension Development via Stimulating Aquaporin Expression and Pulmonary Arterial Smooth Muscle Cells Proliferation.

Pharmacology (2020-01-30)
Yue Wang, Bin Zhong, Qiyong Wu, Tao Zhu, Yong Wang, Ming Zhang
ABSTRACT

The regulatory network of aquaporin (AQP) 1 and renin-angiotensin-aldosterone (ALDO) system are not quite clear in pulmonary arterial hypertension (PAH). Thus, we explored the role of AQP1, ALDO and spirolactone (SP) in the PAH animal model and pulmonary arterial smooth muscle cells (PASMCs). PAH rat model was established by monocrotaline (MCT) via intraperitoneal in SD rat. Hemodynamic measurement was conducted via the external jugular vein cannula. PASMCs were extracted from normal SD rat and cultured in SmGM medium. α-Actin expression was identified by immunocytochemistry. Protein levels were assessed by Western blot. Cell viability was assayed using the MTT method. Apoptosis rate was evaluated by flow cytometry. ALDO level was measured by ELISA. SP decreased AQP1 and β-catenin expressions in PAH rat model induced successfully by MCT. Moreover, ALDO increased AQP1 expression and cell viability in PASMCs, which were extracted from rat and identified by α-actin expression. AQP1 downregulation decreased β-catenin expression, and SP lowered AQP1 and β-catenin expressions elevated by ALDO in PASMCs. SP offset ALDO's effect on the upregulation of cell viability as well as AQP1 and β-catenin expressions in PASMCs. In addition, AQP1 downregulation and SP have a negative effect on Ki-67 and proliferating cell nuclear antigen expressions as well as cell viability after ALDO treatment in PASMCs. ALDO might contribute to PAH development via stimulating AQP1 expression and PASMCs proliferation. However, SP could be considered an effective drug regulating PASMCs proliferation through modulating AQP1 and β-catenin expressions in PAH.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
2-Phenylindole, technical grade, 95%
Pricing and availability is not currently available.
Roche
Cell Proliferation Kit I (MTT)
Pricing and availability is not currently available.
Sigma-Aldrich
SIGMAFAST Protease Inhibitor Cocktail Tablets, EDTA-Free, for use in purification of Histidine-tagged proteins
Pricing and availability is not currently available.
Monocrotaline, phyproof® Reference Substance
Pricing and availability is not currently available.
SAFC
Hanks′ Balanced Salt solution, HBSS Modified, with calcium, with magnesium, without phenol red, liquid, suitable for cell culture
Pricing and availability is not currently available.
SAFC
Hanks′ Balanced Salt solution, HBSS Modified, with phenol red, without calcium, without magnesium, liquid
Pricing and availability is not currently available.
Sigma-Aldrich
Bicinchoninic Acid Kit for Protein Determination, for 200-1000 μg/ml protein
Pricing and availability is not currently available.
Sigma-Aldrich
RIPA Buffer
Pricing and availability is not currently available.
Sigma-Aldrich
Collagenase type I, The collagenase type I (from Clostridium histolyticum) is a crude collagenase preparation that can be used for the isolation of primary cells or for tissue dissociation by enzymatic means.
Pricing and availability is not currently available.