【作者】 李洁；

【导师】 李兵；

【作者基本信息】 重庆医科大学 ， 耳鼻咽喉科学， 2012， 硕士

【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（P<0.05）。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（p>0.05）。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（p<0.05）,RH组与UC组平均灰度值比较无明显差异（p>0.05）。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。更多还原

【Abstract】 Objective：To make a chronic intermittent hypoxia (CIH) Sprague-Dawley（SD）rats model, as a stable and reliable experimental platform for the study ofthe Pathophysiology in sleep disorder breathing. To study the effects oforthobiosis environment, chronic intermittent hypoxia and removal ofhypoxia on oxidative stress system and aorta endothelial cell inSprague-Dawley rats, explore the possible mechanism of oxidative stresssystem disorder and the injury of aorta endothelial cell in patients with sleepdisorder breathing，and to provide a guidance for the clinical treatment ofpatients.Method：Twenty-four sex maturation rats were divided as random number intothree groups：unhandled control group (feeding for4weeks in normalenvironment, UC group), chronic intermittent hypoxia group (feeding for4weeks in chronic intermittent hypoxia cabin, CIH group), reoxygenationgroup(feeding for4weeks in normal environment after4weeks in CIH, RH group).Tested the activity of superoxide dismutase (SOD),malondialdehyde (MDA) and nitric oxide (NO) in blood serum of eachgroup after animal model accomplished. Killed rats by depletion method，dislodged rat’saortaforhematoxylinand eosin（HE）staining, observed theexpression of endothelial nitric oxide synthase (eNOS) byimmunohistochemisty.Results：1．Compared with UC group, the activities of SOD and NO weresignificantly lower in CIH group, but MDA was significantly higher. Thesedifferences were statistically significant (all of P<0.05respectively). Therewere no significantly differences in the weigh of MDA、SOD、NO betweenUC group and RH group (P>0.05).2．HE staining of the aorta: In light microscope the UC group showedaorta endothelial integrity, continuous, smooth and non-shedding ofendothelial cells；tunica intima，tunica media and tunica adventitia clear。Compared with UC group, in the CIH group, endothelial cells part off,aortic intima incomplete, partial smooth muscle cell of tunica media hyperproliferation，arrangement disorder, few smooth muscle cyrto to the tunicaintima. The aorta cells of RH group appeared similar to that of UC group inlight microscope, tunica intima cells full, smooth muscle elastic fibersarranged regularity, and no significant convex in the tunica media.3．Immunohistochemisty of the aorta: we found that brown granules were in aortic endothelialcell’s cytoplasm in UC group and RH group, dyewas lighter and brown granules was less in the endothelial of CIH group.Semi-quantitative analysis of immunohistochemical results: compared withUC group, the average gray level value was significantly below in CIHgroup (p<0.05), and there was no significant difference of average graylevel value between the UC group and RH group (p>0.05).Conclusion：1．Chronic intermittent hypoxia could activate the oxidative stresssystem， reoxygenation after hypoxia can make it back to normal levels.2．Chronic intermittent hypoxia could injure the aortal endothelialcell，and reduce the expression of nitric oxide synthase. Reoxygenationcould make the action of endothelial cell back to the normal levels.更多还原