ABSTRACT
Objective To investigate the molecular mechanism of Ezrin in promoting human lung cancer metastasis.Methods The expression of Ezrin in lung cancer cell lines was detected by using RT-PCR and western blot;the cell scratch test was used to detect the effect of Ezrin on the migration ability of lung cancer cells;the mechanism of Ezrin for regulating L1CAM was detected by western blot.Results The western blot detection results showed that compared with the low metastatic lung cancer cell line H460 and EBC-1,the expression of Ezrin protein the high metastatic lung cancer cell line 95D and PC9 were higher (P＜0.05).After silencing the expression of Ezrin in 95D cells (siEzrin-95D) by using the genetic method,the cell scratch test showed that the migration ability of siEzrin-95D cells was significantly weakened compared with 95D cells (P＜0.05).Compared with 95D cells and H460 cells,after genetically silencing the Ezzin expression in 95D and H460 cells,the L1CAM expression was significantly down-regulated (P＜0.05).Conclusion Ezrin promotes lung cancer metastasis possibly by regulating the expression of L1CAM.
ABSTRACT
Objective To investigate the anti-inflammatory effect of penehyclidine hydrochloride on oleic acid-induced acute respiratory distress syndrome (ARDS) in rats.Methods According to randomize number table method, 30 adult male Sprague-Dawley (SD) rats were divided into control group, model group and treatment group. Catheters were placed in femoral vein in each group. The control group was injected with 1.1 mL/kg physiological saline; the model group was injected with 0.1 mL/kg oleic acid and then injected with 1.0 mL/kg normal saline to establish ARDS model; the treatment group was injected with 0.1 mL/kg oleic acid and then injected with 1.0 mL/kg penehyclidine hydrochloride. At 3 hours after the model was established, blood gas analysis was carried out in each group, oxygenation index (PaO2/FiO2) was calculated, and the levels of serum interleukins (IL-1, IL-6) were measured by enzyme linked immunosorbent assay (ELISA). Rats were sacrificed to harvest lung tissue, and the lung wet/dry ratio (W/D) was calculated; the morphological changes of lung tissue was observed under microscope; the superoxide dismutase (SOD) activity and malondialdehyde (MDA) levels were detected by colorimetry; the expression of tumor necrosis factor-α(TNF-α) in lung tissue was detected by immunohistochemical method. The correlations between lung W/D ratio and various indicators were analyzed using Pearson correlation test.Results Compared with the control group, PaO2/FiO2 in model group was significantly decreased, lung W/D ratio was significantly increased, serum IL-1, IL-6 levels and lung tissue MDA content were significantly increased, lung tissue SOD activity was significantly decreased; the alveolar space was filled with exudate, neutrophils and red blood cells, and there were obvious edema and broadening in pulmonary interstitial and alveolar under light microscope; the type Ⅱ alveolar epithelial cells were partly destroyed, accompanied by lamellar body degeneration and emptying phenomenon under electron microscope; and immunohistochemistry showed that the positive expression of TNF-α in lung tissue was significantly increased. Compared with the model group, PaO2/FiO2 in the treatment group was significantly improved [mmHg (1 mmHg = 0.133 kPa): 204.42±31.61 vs. 113.91±47.78,P < 0.05], the lung W/D ratio was significantly decreased (5.80±0.44 vs. 6.82±0.59,P < 0.01), serum IL-1, IL-6 levels and lung tissue MDA content were significantly decreased [IL-1 (μg/L): 18.38±0.28 vs. 20.04±0.39, IL-6 (μg/L): 12.64±0.67 vs. 14.28±1.33, MDA (nmol/mg): 3.95±0.28 vs. 5.17±0.29, allP < 0.05], the activity of SOD in lung tissue was significantly increased (U/mg: 48.75±2.41 vs. 45.09±1.69,P < 0.01), histological and pathological changes were significantly reduced, and the positive expression of TNF-α in lung tissue was significantly reduced [positive cell rate: (25.80±3.44)％ vs. (38.82±3.59)％,P < 0.01].Conclusion Penehyclidine hydrochloride can effectively improve the oxygenation, alleviate lung injury and reduce pulmonary edema in oleic acid induced ARDS rat by decreasing lung W/D ratio, inhibiting oxidative stress and inflammatory response.