Effects of hypoxia on the function of smooth muscle cells from different orders of pulmonary arteries

doi:10.3969/j.issn.1006-7795.2014.02.018

Abstract

Abstract: Objective To explore the effects of hypoxia on the proliferation and migration of rat pulmonary artery smooth muscle cells(PASMCs) isolated and cultured from different orders of pulmonary arteries. Methods The pulmonary artery branches were gently isolated from male Sprague-Dawley rats(250-350 g) from Capital Medical University and eventually cut into three groups according to the vascular grading: the proximal pulmonary arteries(pulmonary trunk and the first order branches of the intrapulmonary arteries), the middle pulmonary arteries(second to third order intrapulmonary arteries), and the distal pulmonary arteries(forth to fifth order intrapulmonary arteries). Primary PASMCs were cultured from the pulmonary arteries and α-smooth muscle actin(α-SMA) in the cells was identified by immunofluorescence. PASMCs were cultured under normoxia(DMEM+2%FBS, 5%CO₂, 21%O₂, 37 ℃) and hypoxia(DMEM+2%FBS, 5%CO₂, 3%O₂, 37 ℃). Cell proliferation was quantified by using 0.5 g/L methyl thiazolyl tetrazolium(MTT) assay. Western blotting was used for detecting the protein expression level of proliferating cell nuclear antigen(PCNA) in PASMCs. Wound healing test was explored to assess the migration function of the cells. Results Cells isolated from different orders of pulmonary arteries from rats tended to be long spindle and grew in the "peak-valley" mode under microscope. Immunofluorescence results showed that all of the three groups of cells are positively express α-SMA which is a marker of smooth muscle cells. Under physiological condition(normoxic), three groups of PASMCs have the similar ability of proliferation and migration. While after incubation under hypoxia(3%O₂, 48 h), all cells from the three groups showed increased proliferation ability(assessed by MTT and PCNA expression) as well as the migration ability. Moreover, compared to the proximal pulmonary arteries, PASMCs from the middle and distal pulmonary arteries showed much more sensitive to hypoxia stimulus. Conclusion These results suggest that PASMCs from middle and distal pulmonary arteries have higher ability of proliferation and migration than those from proximal under hypoxia condition. This will help to give us some information about the possible mechanism of different responses of different orders of pulmonary arteries under the same pathophysiological stimulus.

Key words: pulmonary artery smooth muscle cells, hypoxia, proliferation, migration

CLC Number:

R332

Guo Huan, Wang Wang, Sun Ran, Liu Jie, Wang Jun. Effects of hypoxia on the function of smooth muscle cells from different orders of pulmonary arteries[J]. Journal of Capital Medical University, 2014, 35(2): 231-236.

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URL: https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/10.3969/j.issn.1006-7795.2014.02.018

https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/Y2014/V35/I2/231

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