| Objectives through the establishment of intermittent hypoxia in rat model of PERK/e IF2 a pathway related factors in p-perk, chop in different time of intermittent hypoxia in rat lung tissue expression and the protective effect of edaravone on p-perk, expression of chop.Methods 90 male Wistar rats, were randomly divided into control group( UC group), intermittent hypoxia group(IH group) and edaravone group(ED group). Each group was subdivided into 3D, 7d, 14 d, 21 d, 28 d of the five sub groups, each sub group with 6 rats in the subjects of the experiment. Every day at the same time, ED group of rats by intravenous injection of Edaravone Injection(3mg/Kg), 30 min and IH group rats were placed in the organic glass box feeding together. First given nitrogen until hypoxia(low oxygen concentration to 5% 30s), and then filled with air and oxygen 40 s the concentration of recovery 21%, continue to inject air 50 s, maintain the oxygen concentration in the 21%. above as a cycle time is 2min, the oxygen concentration in hypoxia box formed between 5%~21% periodically, caused by intermittent hypoxia; UC group of organic glass daily at the same time and in the same IH group feeding box, continuous injection P-perk, comparative observation method of dyeing, again clean, be fast exposure time of 2 min into the air, oxygen concentration remained at 21%. According to the design were rats, in the heart and lungs at room temperature. Heparin saline through the left ascending aortic heart blood, remove the lungs with physiological saline twice, the left lung immersed in 4% paraformaldehyde fixed and paraffin embedded sections were used for HE staining and immunohistochemistry method staining; the right lung tissue preserved in- 80 DEG C refrigerator for RT-PCR was used to detect the chop m RNA.HE UC group of IH group and ED group rats lung tissue pathology morphological changes; Immune Histochemistry method was used to detect the chop protein expression by RT-PCR method for detection of three groups of rat lung tissue of chop m RNA expression.Results 1. using HE staining to observe the morphological changes in lung tissue of rats under optical microscope: significant changes in UC rats lung tissue morphology was compared with UC group, IH group and ED group were with prolonged intermittent hypoxia time appeared pulmonary edema, alveolar septum, part of alveolar fusion with balloon the cavity is larger, alveolar hemorrhage and exudation of visible protein, changes of capillary bed visible inflammatory cell infiltration in pathology, compared with group IH, the pathological changes of the lung tissue of ED rats with light.2. p-PERK immunohistochemical method to detect the rat lung tissue, the expression of CHOP protein: UC group at different time p-PERK, significant changes in the expression of CHOP protein was observed; compared with UC group, IH group and ED group in 3D, 7d, 14 d, 21 d, 28 d and p-PERK at each time point, the expression of CHOP protein were significantly increased(P<0.05), and with the prolonged time of intermittent hypoxia, the expression increased trend P-perk in potential, and IH group compared to the ED group at each time point in rats lung tissue, chop protein expression were decreased( P<0.05). RT-PCR was used to detect the chop m RNA content: UC group at each time point comparison of chop m RNA content had no obvious change; and UC group compared to the IH group and ED groups in 3D, 7d, 14 d, 21 d, 28 d at each time point of chop m RNA expression increased significantly(P<0.05), and with the intermittent hypoxia time prolonged, chop m RNA expression showed a gradual increasing trend; and the IH group compared to the ED group rats lung tissue at each time point of chop m RNA expression were decreased( P<0.05). 4, correlation analysis: p-perk and chop protein expression had positive correlation(r = 0.837, P<0.01 with statistical significance.Conclusions 1 Intermittent hypoxia induced lung injury in rats, and PERK/e IF2 a pathway related factor p-PERK ?CHOP expression in lung tissue. 2 Edaravone reduces p-PERK in lung tissue of rats with intermittent hypoxia, the expression of CHOP and the degree of lung injury in rats. |
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