Objective: To study the effects of 1-(2, 6-dimethylphenoxy)-2-(3, 4-dimethoxyphenylethylamino) propane hydrochloride (DDPH) on proliferation and immunophenotypes of newborn rat pulmonary vascular pericytes (PCs) induced by hypoxic endothelial cell conditioned medium (HECCM) from porcine pulmonary arteries.
Methods: The PCs were divided into 4 groups according to the endothelial cell conditioned medium (ECCM): normoxic ECCM (NECCM) group, NECCM + DDPH group, HECCM group and HECCM + DDPH group. Cell culture, immunocytochemical staining, image analysis and flow cytometry were used to investigate the effects of HECCM and DDPH on the expression of alpha-smooth muscle actin (alpha-SM-actin) and proliferating cell nuclear antigen (PCNA) and cell cycle in PCs.
Results: The alpha-SM-actin antigen in the PCs in HECCM group was more positively expressed than that in the other three groups, but CD(34) antigen and S-100 antigen were negatively expressed. The expression of alpha-SM-actin antigen, CD(34) and S-100 was positive in the groups of NECCM, NECCM + DDPH and HECCM + DDPH. The expression of alpha-SM-actin and PCNA in HECCM group was 1.32 times (F = 11.09, P = 0.000 1) and 1.24 times (F = 2.89, P = 0.025 7) that in NECCM group, 1.30 times (F = 3.65, P = 0.007 0) and 1.21 times (F = 2.63, P = 0.041 4) that in HECCM + DDPH group, respectively; The percentage of the cells in the G(0)-G(1) phase in the HECCM group was lower by 11.7% and 9.1%, in S phase higher by 5.6% and 4.2%, in G(2)-M phase higher by 6.1% and 4.9% than that in the groups of NECCM, HECCM + DDPH, respectively. The inhibitory rates of DDPH on the increased alpha-SM-actin and PCNA syntheses in PCs induced by HECCM were 23.4% and 17.1% respectively. The inhibitory rate on the increased PCs from G(0)-G(1) phase to S phase was 8.3%.
Conclusion: DDPH might directly inhibit PCs from proliferation and immunophenotypical transformation of smooth muscle-like cells induced by HECCM.