本文選題：高濃度氧 + 肺損傷��； 參考：《瀘州醫(yī)學(xué)院》2013年碩士論文

【摘要】：目的研究DRP1（dynamin related protein1）和OPA1（optic atrophy1)在高體積氧誘導(dǎo)早產(chǎn)大鼠肺損傷時細(xì)胞凋亡中的作用。 方法1.以早產(chǎn)大鼠肺組織細(xì)胞為研究對象。隨機(jī)將早產(chǎn)Wistar大鼠48只分為高氧組和對照組，高氧組吸入95%的氧（建立高氧肺損傷模型）,對照組吸入21%的氧的常壓空氣。相同常規(guī)喂養(yǎng)1d、3d、7d分批離斷早產(chǎn)鼠頸放血處死后取肺組織做石蠟切片。2.檢測指標(biāo)：采用蘇木精-伊紅（hematein eosin HE）染色在光鏡下觀察早產(chǎn)鼠肺組織的病理形態(tài)學(xué)變化；免疫組織化學(xué)鏈霉菌抗生物素蛋白-過氧化物酶連結(jié)法（streptavidin-peroxidase SP）觀察與線粒體形態(tài)變化相關(guān)的蛋白：動力相關(guān)蛋白(dynamin related protein1DRPl)和視神經(jīng)萎縮癥蛋白(optic atrophyOPAl)在各組早產(chǎn)大鼠肺組織的表達(dá)和分布及其比值變化；采用原位末端轉(zhuǎn)移酶標(biāo)記技術(shù)（terminal-deoxynucleotidyl transferase mediated nick endlabeling TUNEL）評價各組早產(chǎn)大鼠肺組織細(xì)胞凋亡情況。3.統(tǒng)計學(xué)處理：應(yīng)用SPSS16.0軟件，所有數(shù)據(jù)以x±s表示，組間比較采用單因素方差分析，兩組比較進(jìn)行配對t檢驗和采用LSD法檢驗（a=0.05），指標(biāo)間的關(guān)系采用直線相關(guān)分析進(jìn)行處理,P0.05為差異有統(tǒng)計學(xué)意義。 結(jié)果1.高氧組早產(chǎn)大鼠肺組織細(xì)胞可見典型的肺損傷病理形態(tài)學(xué)改變。在光鏡下觀察，對照組大鼠肺組織細(xì)胞緊密排列多角扁平狀，呈鵝卵石樣改變,透明度好，胞質(zhì)中顆粒稀少。高氧組大鼠肺組織細(xì)胞數(shù)量隨時間（1d、3d、7d）較對照組顯著減少，細(xì)胞形態(tài)改變不規(guī)則，肺泡結(jié)構(gòu)破壞明顯，透光度減弱，，較多空泡、脂肪小滴、顆粒聚集物出現(xiàn)在胞質(zhì)中，細(xì)胞間隙變大，細(xì)胞碎片大量填充細(xì)胞間，間隔增粗肺泡腔變小。2.免疫組織化學(xué)法測定DRP1表達(dá)：高氧組與對照組相比，高氧組細(xì)胞胞質(zhì)中DRPl表達(dá)顯著升高，差異有統(tǒng)計學(xué)意義(p0．05)。DRP1在高氧組平均光密度(Mean±SD)1d(1634.57±108.85),3d(8008.14±530.26),7d(13132.32±594.57);DRP1在對照組平均光密度值(Mean±SD)1d(789.86±58.95),3d(772.31±42.82),7d(770.39±67.8)。3.免疫組織化學(xué)法測定OPAl表達(dá)：高氧組與對照組相比,高氧組細(xì)胞胞質(zhì)中OPA1表達(dá)顯著降低，差異有統(tǒng)計學(xué)意義(p0.05)。OPA1在高氧組平均光密度值(Mean±SD)1d(3126.91±264.91),3d(1399.81±268.62),7d(579.76±99.20);OPA1在對照組平均光密度值(Mean±SD)1d(3935.42±263.99),3d(3764.15±281.53),7d(3907.80±231.34)。4.與對照組相比,高氧組DRP1/OPA1表達(dá)的比值呈時間依賴性增高，差異有統(tǒng)計學(xué)意義(p0．05):高氧組DRP1/OPA1的比值表達(dá)(Mean±SD)1d(0.52±0.01),3d(5.72±0.05),7d(22.68±0.07);對照組DRP1/OPA1的比值表達(dá)(Mean±SD)1d (0.200±0.01),3d(0.205±0.01),7d（0.197±0.01）。5.對照組中可見少量黃色或棕黃色的TUNEL陽性細(xì)胞，高氧組可見大量TUNEL陽性細(xì)胞出現(xiàn)在支氣管上皮細(xì)胞和肺泡上皮細(xì)胞中。高氧暴露時間越長，TUNEL陽性細(xì)胞越多，細(xì)胞凋亡指數(shù)增高,細(xì)胞凋亡隨高氧暴露時間有增加趨勢:高氧組凋亡指數(shù)1d(26.39±1.45),3d(43.87±1.62)，7d(56.34±1.56)；對照組凋亡指數(shù)1d(14.54±1.88),3d(14.68±2.01),7d(14.45±1.75)。6.在高氧組，DRP1/OPA1比值表達(dá)與細(xì)胞凋亡指數(shù)相比，呈顯著正相關(guān)（r=0.725，P0.01）。 結(jié)論1.高氧暴露導(dǎo)致早產(chǎn)鼠肺組織產(chǎn)生急性肺損傷病理學(xué)改變，這一形態(tài)學(xué)改變表現(xiàn)出與高氧暴露有時間依賴性。2.高氧暴露可致新生大鼠肺組織細(xì)胞凋亡，其機(jī)制通過提高DRP1和降低OPA1的表達(dá)水平致線粒體融合分離失衡使線粒體片段化加強(qiáng)促進(jìn)細(xì)胞凋亡，參與高氧肺損傷。3.DRP1/OPA1蛋白比值表達(dá)增高變化與高氧暴露有時間依賴性，細(xì)胞凋亡指數(shù)隨高氧暴露時間有上升趨勢，DRP1/OPA1比值與細(xì)胞凋亡指數(shù)相比具有顯著正相關(guān)性。
[Abstract]:Objective to study the role of DRP1 (dynamin related protein1) and OPA1 (optic atrophy1) in the apoptosis of lung injury induced by high-volume oxygen in premature rats.
Methods 1. the preterm rat lung tissue cells were studied. 48 Wistar rats were randomly divided into hyperoxia group and control group. The hyperoxia group inhaled 95% oxygen (high oxygen lung injury model), and the control group inhaled 21% oxygen atmospheric pressure. The same routine feeding 1D, 3D, 7d batches of premature rat neck bleeding after the death of the lung tissue to make paraffin cut .2. detection index: the pathological changes of lung tissue in premature rats were observed under the light microscope with hematoxylin and eosin (Hematein eosin HE) staining; the protein of immuno histochemical Streptomyces anti biotin peroxidase (streptavidin-peroxidase SP) was used to observe the protein of mitochondrial morphologic changes: dynamic related protein (dyn Amin related protein1DRPl) and optic dystrophy protein (optic atrophyOPAl) expression and distribution of lung tissue in each group of preterm rats and the changes in their ratio. The apoptosis of lung tissue in each group of preterm rats was evaluated by in situ terminal transferase labeling technique (terminal-deoxynucleotidyl transferase mediated Nick endlabeling TUNEL) State.3. statistics: using SPSS16.0 software, all the data were expressed in X + s, and the single factor variance analysis was used in the group. The two groups were compared with the paired t test and LSD test (a=0.05). The relationship between the indexes was treated with linear correlation analysis, and P0.05 was statistically significant.
Results the lung tissue cells in the 1. hyperoxic group showed typical pulmonary pathological changes. Under the light microscope, the lung tissue cells in the control group were closely arranged and flattened, with cobblestone like changes, the transparency was good, and the particles in the cytoplasm were scarce. The number of lung tissue cells in the hyperoxic group was significantly higher than that of the control group (1D, 3D, 7D). Decreased cell morphology changed irregularly, alveolar structure was destroyed obviously, light transmittance was weakened, more vacuoles, fat droplets, granular aggregates appeared in the cytoplasm, cell space became larger, cell fragments were filled with large numbers of cells, and spaced thickening alveolar cavity changed small.2. immunohistochemical method to determine DRP1 expression: hyperoxia group compared with control group, hyperoxia group was fine. The expression of DRPl in cytoplasm was significantly higher, the difference was statistically significant (P0.05).DRP1 in the average light density (Mean + SD) 1D (1634.57 + 108.85), 3D (8008.14 + 530.26), 7d (13132.32 + 594.57), DRP1 in the control group (Mean + SD) 1D (789.86 + 58.95), (772.31 + 42.82), 770.39 + 67.8) immunohistochemical method Al expression: compared with the control group, the expression of OPA1 in the cytoplasm of hyperoxia group decreased significantly, and the difference was statistically significant (P0.05).OPA1 in the hyperoxic group (Mean + SD) 1D (3126.91 + 264.91), 3D (1399.81 + 268.62), 7d (579.76 + 99.20), OPA1 in the control group (3935.42 + 263.99) (3935.42 + 263.99), 3764.15 + 281 .53), 7d (3907.80 + 231.34).4. compared with the control group, the ratio of DRP1/OPA1 expression in the hyperoxic group increased in time dependent, and the difference was statistically significant (P0.05): the ratio of DRP1/OPA1 in the hyperoxic group (Mean + SD) 1D (0.52 + 0.01), 3D (5.72 + 0.05), 7d (22.68 + 0.07), and the ratio of DRP1/OPA1 in the control group (0.200 + 0.01), 0.205 + 0. 01) a small amount of yellow or brown yellow TUNEL positive cells were found in the 7d (0.197 + 0.01).5. control group. A large number of TUNEL positive cells appeared in the bronchial epithelial cells and alveolar epithelial cells in the hyperoxia group. The longer the exposure time of the hyperoxia, the more TUNEL positive cells, the increase of apoptosis index, and the increase of apoptosis with the time of hyperoxia exposure. Potential: apoptosis index of hyperoxia group (26.39 + 1.45), 3D (43.87 + 1.62), 7d (56.34 + 1.56), apoptosis index 1D (14.54 + 1.88), 3D (14.68 + 2.01), 7d (14.45 + 1.75).6. in hyperoxia group. The expression of DRP1/OPA1 ratio was significantly positively correlated with apoptosis index (r=0.725, P0.01).
Conclusion 1. hyperoxia exposure leads to the pathological changes of acute lung injury in preterm rat lung tissue. This morphological change shows that the time dependent.2. hyperoxia exposure can induce apoptosis in the lung tissue of newborn rats. The mechanism by raising the expression of DRP1 and reducing the expression of OPA1 to the mitochondrial fusion separation disequilibrium causes the mitochondrial fragment. The increase of.3.DRP1/OPA1 protein ratio in hyperoxia lung injury is dependent on hyperoxia exposure, and the apoptotic index increases with the exposure time of hyperoxia, and the DRP1/OPA1 ratio has a significant positive correlation with the apoptosis index.
【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】：2013
【分類號】：R722.6

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