【摘要】：目的：探討線粒體ATP敏感鉀通道(Mitochondrial ATP-sensitivePotassium Channel,mitoKATP)對早產(chǎn)鼠高氧肺損傷的保護(hù)作用。 方法：早產(chǎn)Wistar大鼠72只，隨機(jī)分為對照組（n=24）、高氧組（n=24）和二氮嗪組（n=24）。二氮嗪組在高氧暴露前30min，按10mg/kg腹腔注射二氮嗪，其余兩組在相同時(shí)點(diǎn)腹腔注射等量生理鹽水，高氧組和二氮嗪組均置于950mL·L~(-1)氧氣中，對照組置于同一條件的常壓空氣中。分別于空氣或高氧暴露1、3、7d時(shí)收集肺組織，HE染色觀察肺組織病理形態(tài)變化,原位末端標(biāo)記法（TUNEL）檢測肺組織細(xì)胞凋亡率,免疫組化SP法檢測肺組織絲氨酸蛋白酶(Omi/HtrA2)、半胱氨酸蛋白酶-9（caspase-9）和凋亡抑制蛋白（XIAP）的表達(dá)情況,激光共聚集顯微鏡下觀察間接免疫熒光雙染色Omi/HtrA2的胞內(nèi)轉(zhuǎn)位情況。 結(jié)果：對照組d1、d3、d7肺組織無明顯的炎性改變，肺泡結(jié)構(gòu)逐漸發(fā)育成熟；高氧組隨著高氧暴露時(shí)間延長肺組織內(nèi)逐漸出現(xiàn)小血管充血擴(kuò)張，紅細(xì)胞和炎性細(xì)胞滲出，肺泡間隔增厚，間質(zhì)內(nèi)膠原樣物質(zhì)增生，肺泡數(shù)目減少，，肺泡結(jié)構(gòu)簡單化和囊泡化，部分肺組織可見肺泡萎縮和肺不張。與對照組比較,高氧組細(xì)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)明顯增加(P0.01),Omi/HtrA2表達(dá)(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表達(dá)(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胞內(nèi)轉(zhuǎn)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)明顯增高(p0.01),XIAP表達(dá)(d1:5.32±0.37vs6.68±0.40,d3:3.29±0.31vs6.96±0.62, d7:2.40±0.25vs6.65±0.43)減少(p0.01)。與高氧組比較，二氮嗪組肺組織受損得到改善，細(xì)胞凋亡率（d1：19.82±3.16、d3:31.75±2.39、d7:37.93±2.56）減少，差異有統(tǒng)計(jì)學(xué)意義（P0.01），Omi/HtrA2表達(dá)（d1:7.59±0.40、d3:8.37±0.45、d7:9.23±0.27）減少，差異具有統(tǒng)計(jì)學(xué)意義（P0.01），caspase-9表達(dá)（d1:8.31±0.39、d3:10.32±0.50、d7:12.61±0.41）減少,差異具有統(tǒng)計(jì)學(xué)意義(p0.01),Omi/HtrA2胞內(nèi)轉(zhuǎn)位率（d1:18.40±1.90、d3:38.44±0.94、d7:40.04±1.28）明顯減少，差異具有統(tǒng)計(jì)學(xué)意義(p0.01)，XIAP表達(dá)增多（d1:5.83±0.39、d3:4.95±0.16、d7:3.87±0.44），差異具統(tǒng)計(jì)學(xué)意義(p0.01)。 結(jié)論：二氮嗪可能通過開放mitoKATP減少Omi/HtrA2和caspase-9的表達(dá)，減少Omi/HtrA2在細(xì)胞內(nèi)的轉(zhuǎn)位，阻抑細(xì)胞凋亡，從而減輕早產(chǎn)鼠高氧肺損傷。
[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 鹵1.19 d 3: 43.96 鹵1.33 2.76vs13.83 鹵1.33 d 7: 56.78 鹵2.23vs12.56 鹵1.29) was significantly increased (P0.01). The expression of Omi/HtrA2 (d1: 9.72 鹵0.89vs5.06 鹵1.04d3: 11.08 鹵1.73vs5.26 鹵1.90d7: 13.32 鹵1.12vs6.46 鹵1.45) and caspase-9 (d1: 10.13 鹵0.77vs5.55 鹵0.53d3: 12.66 鹵0.61vs5.11 鹵1.27) were detected. D7: 14.58 鹵0.46vs5.05 鹵1.12 increased (p0.01), and the intracellular transposition rate of Omi/HtrA2 (d1: 23.84 鹵2.20vs5.75 鹵0.82d3: 43.04 鹵2.36v8.11 鹵0.94d7: 54.19 鹵2.87vs8.85 鹵0.72) increased significantly (p0.01). The expression of XIAP (d1: 5.32 鹵0.37vs6.68 鹵0.40d3: 3.29 鹵0.31vs6.96 鹵0.62d 7: 2.40 鹵0.25vs6.65 鹵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 鹵3.16 d 3: 31.75 鹵2.39 d: 7 37.93 鹵2.56) was decreased significantly (P0.01). The expression of Omi/HtrA2 (d17.59 鹵0.40d3w 8.37 鹵0.45d7: 9.23 鹵0.27) decreased, the difference was statistically significant (P0.01), the expression of caspase-9 (D1: 8.31 鹵0.39d3w 10.32 鹵0.50), D7: 12.61 鹵0.41, the difference was statistically significant (p0.01), and the intracellular transposition rate of Omi/HtrA2 (d1: 18.40 鹵1.90) was significantly decreased (38.44 鹵0.94d7: 40.04 鹵1.28). The difference was statistically significant (p0.01), XIAP expression increased (d 1: 5.83 鹵0.39 d 34.95 鹵0.16 d 7: 3.87 鹵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.
【學(xué)位授予單位】：瀘州醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R722.6

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