慢性间隙性缺氧对大鼠氧化应激及血管内皮的影响

发布时间：2018-05-21 09:41

本文选题：慢性间隙性缺氧 + 氧化应激　；参考：《重庆医科大学》2012年硕士论文

【摘要】：目的：制作一个可应用于研究正常压力下慢性间隙性缺氧（Chronicintermittent hypoxia, CIH）的SD大鼠动物模型，为研究睡眠呼吸疾病特征性的间隙性缺氧—复氧中的病理生理过程建立一个稳定可靠的实验平台。通过研究正常生活条件、慢性间隙性缺氧以及缺氧后复氧的情况下对SD大鼠的氧化应激系统以及主动脉血管内皮系统的影响，探索睡眠呼吸疾病患者中氧化应激系统紊乱与主动脉内皮细胞损伤的可能机制，为临床疾病的治疗提供思路。方法： SD性成熟大鼠24只按照随机数字法分为三组即：空白对照组（Unhandled control group）（正常生活环境饲养四周，UC组）、慢性间隙性缺氧组（Chronic intermittent hypoxia group）（慢性间隙性缺氧舱间缺氧四周，CIH组）以及复氧组（Removalof hypoxia group）（间隙性缺氧四周后再正常饲养四周，共八周，RH组）；所有大鼠造模成功后测定各组血清中氧化应激指标：丙二醛（Malondialdehyde, MDA）、超氧歧化物（Superoxide dismutase, SOD）、一氧化氮（Nitric oxide, NO）水平。放血法处死大鼠，取出主动脉进行HE染色，并用免疫组化方法观察内皮型一氧化氮合酶（Endothelialnitric oxide synthase, eNOS）的表达情况。结果： 1、与UC组比较，CIH组血清的氧化指标MDA含量显著升高，SOD、NO含量明显降低，差别有统计学意义（P0.05）。RH组与UC组比较，MDA、NO、SOD含量均无明显差异，差别无统计学意义（p0.05）。 2、主动脉HE染色：光镜下见UC组血管内皮完整、连续、光滑，内皮细胞无脱落，内膜、中膜、外膜分界清楚。CIH组与UC组比，内皮细胞部分脱落，主动脉内膜不完整，，部分中膜平滑肌增生，排列紊乱，少部分平滑肌向内膜层突起。RH组镜下形态基本同UC组，内膜细胞完整，平滑肌弹力纤维层排列整齐，中膜层未见明显突起。 3、主动脉免疫组化发现： UC组及RH组均可见棕黄色颗粒位于主动脉内皮细胞胞浆中，CIH组内皮染色较浅，棕黄色颗粒较少。免疫组化半定量分析结果：CIH组的平均灰度值明显低于UC组（p0.05）,RH组与UC组平均灰度值比较无明显差异（p0.05）。结论： 1、慢性间隙性缺氧中SD大鼠氧化应激系统被激活，缺氧后复氧能使其基本恢复至正常水平。 2、慢性间隙性缺氧过程损伤大鼠主动脉内皮细胞，并导致内皮型一氧化氮表达减少。复氧可以使内皮功能恢复至正常水平。
[Abstract]:Objective: A SD rat model of chronic interstitial hypoxia (CIH) under normal pressure was made to establish a stable and reliable experimental platform for the study of the pathophysiological process in the characteristic intermittent hypoxia reoxygenation of sleep respiratory diseases. By studying the effects of normal living conditions, chronic intermittent hypoxia and reoxygenation after hypoxia on oxidative stress system and aortic vascular endothelial system in SD rats, To explore the possible mechanism of oxidative stress system disorder and aortic endothelial cell injury in patients with sleep apnea disease, and to provide ideas for the treatment of clinical diseases. Methods: Twenty-four SD sexual mature rats were divided into three groups according to the random number method: the blank control group was unhandled control group (normal living environment was fed for four weeks) and the chronic interstitial hypoxia group was chronic intermittent hypoxia group (chronic intermittent hypoxia for four weeks). In the reoxygenation group, the hypoxia group was fed normally for four weeks after intermittent hypoxia. The levels of serum oxidative stress (MDA), malondialdehyde (MDA), malondialdehyde (MDAA), superoxide dismutase (SOD), nitric oxide (no) were measured in all rats. The rats were killed by bloodletting, the aorta was removed for HE staining, and the expression of Endothelia oxide synthase, eNOS) was observed by immunohistochemical method. Results: 1.Compared with UC group, the content of serum MDA increased significantly, and the content of MDA decreased significantly. There was no significant difference between RH group and UC group, but there was no significant difference between RH group and UC group. 2HE staining of aorta: under light microscope, the vascular endothelium in UC group was intact, continuous, smooth, endothelium cells were not exfoliated, intima, media and adventitia boundaries were clear. Compared with UC group, the endothelial cells in CIH group were partially exfoliated, and the aortic intima was incomplete. Some of the medial smooth muscle proliferated and arranged disorderly. A few of the smooth muscle cells were similar to the UC group under microscope. The intimal cells were intact, the smooth muscle elastic fiber layer was arranged neatly, and there was no obvious protuberance in the middle layer. 3. The results of immunohistochemistry of aorta showed that brown granules were found in endothelial cytoplasm of aortic endothelial cells in UC group and RH group. In CIH group, the staining of endothelium was lighter and the brown granules were less. The results of semi-quantitative immunohistochemical analysis showed that the average gray value of the group of CIH was significantly lower than that of the group of UC (P 0.05) and that of the group of UC was not significantly different from that of the group of UC (P 0.05). Conclusion: 1. The oxidative stress system of SD rats was activated during chronic interstitial hypoxia. 2, chronic interstitial hypoxia injures aorta endothelial cells and reduces the expression of endothelial nitric oxide (no). Reoxygenation can restore endothelial function to normal level.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R766.4

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