The efficacy of aminophylline on the apoptosis of human trachea smooth muscle cells in vitro / 中国药理学通报

ABSTRACT

AIM To investigate weather and how aminophylline induces human trachea smooth muscle cells (TSMCs) to apoptosis. METHODS Human TSMCs were isolated and cultured in DMEM containing 10% fetal bovine serum. Passage 4～6 cell were used in experiment. The cells were cultured with aminophylline or 8 Br cAMP for 24 or 48 h. Light micros copy and electron microscopy were used to observe morphological change. DNA fragmentation was analyzed by agarose gels electrophoresis. SP Immunohistological staing method was performed to detect the changes of expressions of p53,bcl 2 and bax gene. The apoptosis cell percentage were detected by situ end labeling technique(TUNEL) of fragmental DNA. RESULTS ①Aminophylline or 8 Br cAMP decreased the number of viable cells in time and concentration dependent manner; ② Electron microscopic examination showed nuclear contraction, chromatin condensation and apoptotic bodies formation in aminophylline group or 8 Br cAMP group; ③ Agarose gel electrophoresis of fragmented DNA showed a ladder like pattern; ④ The expression of p53 or bax gene in apoptosis group was significantly higher than in control group, but the expression of bcl 2 gene was lower than in control group; ⑤ The positive rate of TUNEL in aminophylline or 8 Br cAMP group was significantly higher than in control group ( P