Effects of DDPH on HECCM-induced Proliferation and Immunophenotypes of the Pulmonary Vascular Pericytes

摘要

In order 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 induced by hypoxic endothelial cell conditioned medium (HECCM)from porcine pulmonary arteries, the cultured pericytes were divided into 4 groups according to the endothelial cell conditioned medium (ECCM)used: normoxic ECCM (NECCM)group, NECCM+DDPH group, HECCM group and HECCM+DDPH group. Cell culture, immunocytochemical staining, image analysis and flow cytometric method were used to investigate the effects of HECCM and DDPH on the expression of a-smooth muscle actin (α-SM-Actin) antigen, CD34 antigen, S-100 antigen and proliferating cell nuclear antigen (PCNA) and cell cycle in pericytes. The results showed that the α-SM-Actin antigen in the pericytes in HECCM group was stronger positively expressed than in the other three groups, but CD34 antigen and S-100 antigen were negatively expressed. The expression of α-SM-Actin antigen, CD34 antigen and S-100 antigen was positive in the groups of NECCM,NECCM +DDPH and HECCM +DDPH; The expression of α-SM-Actin and PCNA in HECCM group was 1.32 times (P＜0. 01) and 1.24 times (P＜0. 05) that in NECCM group, 1.30 times (P＜0.01) and 1.21 times (P＜0. 05) that in HECCM+DDPH group, respectively. The percentage of the cells in the GO-G1 phase in the HECCM group was lower by 11.7 % and 9. 1 %, in S phase higher by 5.6 % and 4.2 %, in G2-M phase higher by 6.1% and 4.9 % than in the groups of NECCM,HECCM+DDPH, respectively. The inhibitory rate of DDPH on the increased α-SM-Actin and PCNA syntheses in pericytes induced by HECCM were 23.4 % and 17. 1 % respectively. The inhibitory rate on the increased pericytes from GO-G1 phase to S phase was 8. 3 %. These results suggest that DDPH can directly inhibit pericytes from proliferation and immunophenotypical transformation of smooth muscle-like cells induced by HECCM.