Of nitric oxide synthase in the role of anti-tumor cells.doc

 PAGE \* MERGEFORMAT 9 Of nitric oxide synthase in the role of anti-tumor cells Keywords:: alveolar macrophage nitric oxide synthase in tumor Activated macrophages (M) in the anti-tumor process is of great significance. Of nitric oxide (NO) as the activation of the M, one of the cytotoxic effector molecule in the anti-tumor effects play an important role [1]. This study by measuring the lung M in inducible nitric oxide synthase (iNOS) and its product NO levels, and application-specific NOS inhibitor, to observe the lung tumor cell killing effect of M on the effects to investigate the activity of pulmonary M NOS The relationship between tumor mass destruction in order to clarify the lung M in the anti-tumor immunity status. 1 Materials and methods 1.1 Preparation of lung M and nurture body mass 20 ~ 25 g of pure C57 male mice, 32, purchased from Shanghai Sciple - Bikai experimental animal Co., Ltd., were randomly divided into experimental group and control group, 16 rats in each, are After intraperitoneal injection of 2% sodium pentobarbital anesthesia, abdominal aorta bleeding death, the trachea after exposure to the neck, with 37 ℃ saline for bronchoalveolar lavage, each 1 ml, a total of 15 times. The recovered lavage fluid to 1 500 r / min centrifugation 10 min, discard supernatant, containing 10% fetal calf serum minimum essential medium (MEM) dubbed in 1 * 106 / ml of cell suspension. Suspension of cells seeded in 96 well plate, 37 ℃ , 5% CO2 culture 2 h, washing away the non-adherent cells were added to different doses as needed (50,100,150,200 U / ml) of IFN-γ (INFγ ), continued to train 24 h. 1.2 Tumor cell culture supernatant collected will be made sterile mast cell tumor cells P815 (Shanghai Cancer Institute) to 5 * 105 / ml density cultured in RPMI 1640 medium, Chi 37 ℃ , 5% CO2 incubator cultured 48 h, 2 500 r / min centrifugation 20 min, the supernatant, set -20 ℃ to save stand-by. 1.3 iNOS activity was determined by Smith thin-layer chroma