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目的 观察具有抗高血压作用新药 1 (2 ,6 二甲基苯氧基 ) 2 (3,4 二甲氧基苯乙氨基 )丙烷盐酸盐 (DDPH)对低氧内皮细胞条件培养液 (HECCM)诱导的大鼠肺血管周细胞 (PC)增殖和免疫表型转化有无抑制作用 ,为开发具有降压和防止肺血管构型重建新药物提供实验依据。方法 实验根据不同条件培养液分为 4组 :常氧内皮细胞条件培养液 (NECCM)组 ,NECCM +DDPH组 ,HECCM组和HECCM +DDPH组。应用细胞培养、免疫细胞化学染色、图像分析及流式细胞术观察HECCM和DDPH对PC平滑肌肌动蛋白 (α SM actin)和增殖细胞核抗原 (PCNA)的表达及细胞周期的影响。结果 ①HECCM组PCα SM actin呈强阳性表达 ,CD34 和S 10 0呈阴性反应 ,而其它各组PCα SM actin、CD34及S 10 0均呈阳性表达 ;②HECCM组PCα SM actin和PCNA的表达量分别是NECCM组的 1 32倍 (F=11 0 9,P =0 0 0 0 1)和 1 2 4倍 (F =2 89,P =0 0 2 5 7) ,是HECCM +DDPH组的 1 30倍 (F =3 6 5 ,P =0 0 0 70 )和 1 2 1倍 (F =2 6 3,P =0 0 414) ;③HECCM组PC的G0 G1期细胞百分率分别比NECCM组和HECCM +DDPH组低 11 7%和 9 1% ,S期细胞百分率分别高 5 6 %和 4 2 % ,G2 M期细胞百分率分别高 6 1%和 4 9% ;④DDPH对HECCM引起的PCs合成α SM actin和PCNA增高?
Objective To observe the effects of 1, 2,6-dimethylphenoxy-2 (3,4-dimethoxyphenylethylamino) propane hydrochloride (DDPH), a new antihypertensive drug, on hypoxic endothelial cell conditioned medium (HECCM ) Induced rat pulmonary pericytes (PC) proliferation and immunophenotypic transformation without inhibition, to provide experimental evidence for the development of antihypertensive and prevent pulmonary vascular reconstruction of new drugs. Methods The culture medium was divided into 4 groups according to different conditions: normoxic endothelial cell conditioned medium (NECCM), NECCM + DDPH, HECCM and HECCM + DDPH groups. Cell culture, immunocytochemical staining, image analysis and flow cytometry were used to observe the effects of HECCM and DDPH on the expression of α SM actin and proliferating cell nuclear antigen (PCNA) and cell cycle. Results ① The positive expression of PCα SM actin in HECCM group was negative, but the expression of PCα SM actin, CD34 and S 10 0 was negative in CD34 and S 10 0 in other groups. ② The expressions of PCα SM actin and PCNA in HECCM group were The NECCM group was 130 times (F = 11 0 9, P = 0 0 0 0 1) and 1 2 4 times (F = 2 89, P = 0 0 2 5 7) (F = 36.5, P = 0 0 0 70) and 12 1 times (F = 2633, P = 0 0 414). ③ The percentages of cells in G0 G1 phase in HCECCM group were significantly higher than those in NECCM group and HECCM + DDPH group Group were 11 7% and 91% respectively, the percentage of cells in S phase were 56% and 42% respectively, and the percentage of cells in G2 M phase was 61% and 49% respectively; ④ DDPH inhibited the synthesis of α SM actin and PCNA increased?