线粒体ATP敏感钾通道对早产鼠高氧肺损伤的保护作用
发布时间：2018-11-27 13:51
【摘要】：目的：探讨线粒体ATP敏感钾通道(Mitochondrial ATP-sensitivePotassium Channel,mitoKATP)对早产鼠高氧肺损伤的保护作用。 方法：早产Wistar大鼠72只，随机分为对照组（n=24）、高氧组（n=24）和二氮嗪组（n=24）。二氮嗪组在高氧暴露前30min，按10mg/kg腹腔注射二氮嗪，其余两组在相同时点腹腔注射等量生理盐水，高氧组和二氮嗪组均置于950mL·L~(-1)氧气中，对照组置于同一条件的常压空气中。分别于空气或高氧暴露1、3、7d时收集肺组织，HE染色观察肺组织病理形态变化,原位末端标记法（TUNEL）检测肺组织细胞凋亡率,免疫组化SP法检测肺组织丝氨酸蛋白酶(Omi/HtrA2)、半胱氨酸蛋白酶-9（caspase-9）和凋亡抑制蛋白（XIAP）的表达情况,激光共聚集显微镜下观察间接免疫荧光双染色Omi/HtrA2的胞内转位情况。 结果：对照组d1、d3、d7肺组织无明显的炎性改变，肺泡结构逐渐发育成熟；高氧组随着高氧暴露时间延长肺组织内逐渐出现小血管充血扩张，红细胞和炎性细胞渗出，肺泡间隔增厚，间质内胶原样物质增生，肺泡数目减少，，肺泡结构简单化和囊泡化，部分肺组织可见肺泡萎缩和肺不张。与对照组比较,高氧组细胞凋亡率(d1:25.02±4.48vs12.30±1.19,d3:43.96±2.76vs13.83±1.33,d7:56.78±2.23vs12.56±1.29)明显增加(P0.01),Omi/HtrA2表达(d1:9.72±0.89vs5.06±1.04,d3:11.08±1.73vs5.26±1.90,d7:13.32±1.12vs6.46±1.45)、caspase-9表达(d1:10.13±0.77vs5.55±0.53,d3:12.66±0.61vs5.11±1.27,d7:14.58±0.46vs5.05±1.12)增多(p0.01)，Omi/HtrA2胞内转位率(d1:23.84±2.20vs5.75±0.82,d3:43.04±2.36v8.11±0.94,d7:54.19±2.87vs8.85±0.72)明显增高(p0.01),XIAP表达(d1:5.32±0.37vs6.68±0.40,d3:3.29±0.31vs6.96±0.62, d7:2.40±0.25vs6.65±0.43)减少(p0.01)。与高氧组比较，二氮嗪组肺组织受损得到改善，细胞凋亡率（d1：19.82±3.16、d3:31.75±2.39、d7:37.93±2.56）减少，差异有统计学意义（P0.01），Omi/HtrA2表达（d1:7.59±0.40、d3:8.37±0.45、d7:9.23±0.27）减少，差异具有统计学意义（P0.01），caspase-9表达（d1:8.31±0.39、d3:10.32±0.50、d7:12.61±0.41）减少,差异具有统计学意义(p0.01),Omi/HtrA2胞内转位率（d1:18.40±1.90、d3:38.44±0.94、d7:40.04±1.28）明显减少，差异具有统计学意义(p0.01)，XIAP表达增多（d1:5.83±0.39、d3:4.95±0.16、d7:3.87±0.44），差异具统计学意义(p0.01)。 结论：二氮嗪可能通过开放mitoKATP减少Omi/HtrA2和caspase-9的表达，减少Omi/HtrA2在细胞内的转位，阻抑细胞凋亡，从而减轻早产鼠高氧肺损伤。
[Abstract]:Objective: to investigate the protective effect of mitochondrial ATP sensitive potassium channel (Mitochondrial ATP-sensitivePotassium Channel,mitoKATP) on hyperoxia lung injury in preterm rats. Methods: 72 preterm Wistar rats were randomly divided into control group (n = 24), hyperoxia group (n = 24) and diazoxide group (n = 24). The diazazine group was injected intraperitoneally with 10mg/kg 30 min before hyperoxia exposure, and the other two groups were injected with the same amount of saline at the same time. The hyperoxia group and diazazine group were placed in 950mL L ~ (-1) oxygen. The control group was placed in atmospheric air under the same conditions. Lung tissues were collected after air or hyperoxia exposure for 7 days. HE staining was used to observe the histopathological changes of lung tissues. The apoptosis rate of lung tissue was detected by in situ end labeling (TUNEL). The expression of serine protease (Omi/HtrA2), cysteine protease-9 (caspase-9) and apoptosis-inhibiting protein (XIAP) in lung tissue was detected by immunohistochemical SP method. The intracellular translocation of Omi/HtrA2 by indirect immunofluorescence double staining was observed under laser co-aggregation microscope. Results: in the control group, there were no obvious inflammatory changes and alveolar structure matured gradually. With the extension of hyperoxia exposure time, small blood vessel congestion and dilatation, exudation of erythrocytes and inflammatory cells, thickening of alveolar septum, proliferation of collagenous substance in interstitium, decrease of alveolar number, simplification of alveolar structure and vesicle in hyperoxia group. Alveolar atrophy and atelectasis were seen in some lung tissues. Compared with the control group, the apoptosis rate of hyperoxia group (d 1: 25.02 卤1.19 d 3: 43.96 卤1.33 2.76vs13.83 卤1.33 d 7: 56.78 卤2.23vs12.56 卤1.29) was significantly increased (P0.01). The expression of Omi/HtrA2 (d1: 9.72 卤0.89vs5.06 卤1.04d3: 11.08 卤1.73vs5.26 卤1.90d7: 13.32 卤1.12vs6.46 卤1.45) and caspase-9 (d1: 10.13 卤0.77vs5.55 卤0.53d3: 12.66 卤0.61vs5.11 卤1.27) were detected. D7: 14.58 卤0.46vs5.05 卤1.12 increased (p0.01), and the intracellular transposition rate of Omi/HtrA2 (d1: 23.84 卤2.20vs5.75 卤0.82d3: 43.04 卤2.36v8.11 卤0.94d7: 54.19 卤2.87vs8.85 卤0.72) increased significantly (p0.01). The expression of XIAP (d1: 5.32 卤0.37vs6.68 卤0.40d3: 3.29 卤0.31vs6.96 卤0.62d 7: 2.40 卤0.25vs6.65 卤0.43) was decreased (p0.01). Compared with the hyperoxia group, the lung tissue damage in the diazazine group was improved, and the apoptosis rate (d 1: 19.82 卤3.16 d 3: 31.75 卤2.39 d: 7 37.93 卤2.56) was decreased significantly (P0.01). The expression of Omi/HtrA2 (d17.59 卤0.40d3w 8.37 卤0.45d7: 9.23 卤0.27) decreased, the difference was statistically significant (P0.01), the expression of caspase-9 (D1: 8.31 卤0.39d3w 10.32 卤0.50), D7: 12.61 卤0.41, the difference was statistically significant (p0.01), and the intracellular transposition rate of Omi/HtrA2 (d1: 18.40 卤1.90) was significantly decreased (38.44 卤0.94d7: 40.04 卤1.28). The difference was statistically significant (p0.01), XIAP expression increased (d 1: 5.83 卤0.39 d 34.95 卤0.16 d 7: 3.87 卤0.44), the difference was statistically significant (p0.01). Conclusion: diazazine may reduce the expression of Omi/HtrA2 and caspase-9, decrease the translocation of Omi/HtrA2 in cells and inhibit apoptosis by opening up mitoKATP, thus attenuating hyperoxia lung injury in preterm rats.
【学位授予单位】：泸州医学院
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R722.6
本文编号：2361010
[Abstract]:Objective: to investigate the protective effect of mitochondrial ATP sensitive potassium channel (Mitochondrial ATP-sensitivePotassium Channel,mitoKATP) on hyperoxia lung injury in preterm rats. Methods: 72 preterm Wistar rats were randomly divided into control group (n = 24), hyperoxia group (n = 24) and diazoxide group (n = 24). The diazazine group was injected intraperitoneally with 10mg/kg 30 min before hyperoxia exposure, and the other two groups were injected with the same amount of saline at the same time. The hyperoxia group and diazazine group were placed in 950mL L ~ (-1) oxygen. The control group was placed in atmospheric air under the same conditions. Lung tissues were collected after air or hyperoxia exposure for 7 days. HE staining was used to observe the histopathological changes of lung tissues. The apoptosis rate of lung tissue was detected by in situ end labeling (TUNEL). The expression of serine protease (Omi/HtrA2), cysteine protease-9 (caspase-9) and apoptosis-inhibiting protein (XIAP) in lung tissue was detected by immunohistochemical SP method. The intracellular translocation of Omi/HtrA2 by indirect immunofluorescence double staining was observed under laser co-aggregation microscope. Results: in the control group, there were no obvious inflammatory changes and alveolar structure matured gradually. With the extension of hyperoxia exposure time, small blood vessel congestion and dilatation, exudation of erythrocytes and inflammatory cells, thickening of alveolar septum, proliferation of collagenous substance in interstitium, decrease of alveolar number, simplification of alveolar structure and vesicle in hyperoxia group. Alveolar atrophy and atelectasis were seen in some lung tissues. Compared with the control group, the apoptosis rate of hyperoxia group (d 1: 25.02 卤1.19 d 3: 43.96 卤1.33 2.76vs13.83 卤1.33 d 7: 56.78 卤2.23vs12.56 卤1.29) was significantly increased (P0.01). The expression of Omi/HtrA2 (d1: 9.72 卤0.89vs5.06 卤1.04d3: 11.08 卤1.73vs5.26 卤1.90d7: 13.32 卤1.12vs6.46 卤1.45) and caspase-9 (d1: 10.13 卤0.77vs5.55 卤0.53d3: 12.66 卤0.61vs5.11 卤1.27) were detected. D7: 14.58 卤0.46vs5.05 卤1.12 increased (p0.01), and the intracellular transposition rate of Omi/HtrA2 (d1: 23.84 卤2.20vs5.75 卤0.82d3: 43.04 卤2.36v8.11 卤0.94d7: 54.19 卤2.87vs8.85 卤0.72) increased significantly (p0.01). The expression of XIAP (d1: 5.32 卤0.37vs6.68 卤0.40d3: 3.29 卤0.31vs6.96 卤0.62d 7: 2.40 卤0.25vs6.65 卤0.43) was decreased (p0.01). Compared with the hyperoxia group, the lung tissue damage in the diazazine group was improved, and the apoptosis rate (d 1: 19.82 卤3.16 d 3: 31.75 卤2.39 d: 7 37.93 卤2.56) was decreased significantly (P0.01). The expression of Omi/HtrA2 (d17.59 卤0.40d3w 8.37 卤0.45d7: 9.23 卤0.27) decreased, the difference was statistically significant (P0.01), the expression of caspase-9 (D1: 8.31 卤0.39d3w 10.32 卤0.50), D7: 12.61 卤0.41, the difference was statistically significant (p0.01), and the intracellular transposition rate of Omi/HtrA2 (d1: 18.40 卤1.90) was significantly decreased (38.44 卤0.94d7: 40.04 卤1.28). The difference was statistically significant (p0.01), XIAP expression increased (d 1: 5.83 卤0.39 d 34.95 卤0.16 d 7: 3.87 卤0.44), the difference was statistically significant (p0.01). Conclusion: diazazine may reduce the expression of Omi/HtrA2 and caspase-9, decrease the translocation of Omi/HtrA2 in cells and inhibit apoptosis by opening up mitoKATP, thus attenuating hyperoxia lung injury in preterm rats.
【学位授予单位】：泸州医学院
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R722.6
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