高压氧对P53和Caspase-3在SAH后早期脑损伤中表达影响的研究

发布时间：2018-04-24 15:42

  本文选题：蛛网膜下腔出血 + 早期脑损伤　； 参考：《川北医学院》2014年硕士论文

【摘要】：背景与目的：蛛网膜下腔出血（subarachnoid hemorrhage，SAH）是高病死率和高致残率的脑血管疾病之一，分为自发性和外伤性两类。由颅内动脉瘤破裂所致的SAH占所有自发性SAH的比例高达80%左右，又称动脉瘤性蛛网膜下腔出血，其致死率和致残率极高，出血后3天内死亡率可达45%。学者认为出血后的早期脑损伤（early brain injury，EBI）是其高死亡率和致残率的主要因素之一。高压氧治疗（hyperbaric oxygentherapy，HBOT）是在高于一个大气压的环境里吸入100%的氧，目前已广泛应用于多种疾病的治疗，由于SAH患者病情变化快且风险大，高压氧应用于SAH的早期治疗研究极少。本研究采用颈内动脉血管内穿刺法建立大鼠SAH动物模型，观察大鼠脑皮质中P53和Caspase-3的表达情况，以及高压氧干预后大鼠神经行为学、死亡率和脑皮质中P53和Caspase-3的表达变化。 方法：选取体重为280-320g的健康成年雄性SD大鼠100只，，随机分为正常空白对照组10只、假手术组（sham组）30只、蛛网膜下腔出血组（SAH组）30只、高压氧干预的SAH组（HBOT+SAH组）30只；除空白对照组外，其余各组按实验动物处死取标本时间再分为24h、48h、72h三个亚组，各亚组10只。空白对照组不做任何特殊处理，常规饲养3天后麻醉、灌注、取脑组织标本保存；SAH组和HBOT+SAH组均用颈内动脉血管内穿刺法建模，建模成功后除HBOT+SAH组大鼠待麻醉苏醒后行高压氧治疗外，其余饲养环境和术后护理均一样；sham组建模过程与SAH组相同，但不刺破血管。sham组、SAH组和HBOT+SAH组分别于建模成功后24h、48h、72h记录各亚组大鼠神经行为学评分和死亡率，然后麻醉、灌注、取各亚组大鼠脑组织保存。采用免疫组织化学法检测大鼠脑皮质中P53和Caspase-3的表达情况。 使用Motic Image Advance3.2图像分析系统对免疫组织切片进行分析，计算其阳性细胞百分比，Microsoft office excel2003软件录入数据，正态分布计量资料以S表示。采用spss19.0软件进行数据分析，比较前进行正态性和方差齐性检验，两组间比较采用t检验，各亚组间两两比较采用SNK法（q检验），P＜0.05认为有统计学意义。 结果： 1.死亡率：空白对照组和假手术组大鼠死亡率均为0，两组比较无统计学意义；SAH组和HBOT+SAH组总死亡率分别为40%和16.67%，两组死亡率比较有统计学意义（P＜0.05）；SAH组和HBOT+SAH组在24h内死亡率分别为23.33%和13.33%，24h内两组死亡率比较无统计学差异（P＞0.05）。 2.神经行为学评分：SAH组与HBOT+SAH组在24h、48h两个亚组比较无差异（P＞0.05）；但HBOT+SAH组中72h亚组的评分明显低于SAH组中72h亚组，差异有统计学意义（P＜0.05）；SAH组中三个亚组间两两比较无明显差异（P＞0.05）；HBOT+SAH组中三亚组间两两比较：24h亚组与48h亚组间比较无统计学意义（P＞0.05）；24h亚组与72h亚组、48h亚组与72h亚组比较均有统计学意义（P＜0.05）。 3.P53在大脑皮质中阳性表达为细胞核呈棕褐色或棕黄色颗粒，并伴有少量胞质表达。空白对照组和假手术组均有少量表达，但两组比较无统计学意义（P＞0.05）；SAH三亚组间两两比较，仅24h与48h亚组P53阳性表达细胞百分比无差异；SAH各亚组比对应sham各亚组阳性表达明显增多，有统计学意义（P＜0.05）；HBOT+SAH组三亚组间两两比较均有统计学意义；P53在HBOT+SAH组48h、72h亚组表达低于相对应的SAH两亚组，差异有统计学意义（P＜0.01）；HBOT+SAH-24h组与SAH-24h组比较无差异（P＞0.05）。 4.Caspase-3在胞质上表达，阳性表达为棕黄色颗粒。空白对照组和假手术组均处于正常低水平表达，差异无统计学意义（P＞0.05）；SAH各亚组比对应sham各亚组阳性表达明显增多，有统计学意义（P＜0.05）；SAH组各时间亚组间两两比较均有差异；HBOT+SAH各亚组间两两比较差异有统计学意义。Caspase-3在SAH组与HBOT+SAH组比较：在24h、48h、72h亚组中均有统计学意义（P＜0.05） 结论： 1.大鼠SAH后早期脑损伤与脑皮质中P53和Caspase-3的表达增加有关。 2.高压氧治疗能降低大鼠SAH后早期（72小时内）的总死亡率，并在一定程度上改善神经功能，对治疗EBI有一定作用，但对降低24h内大鼠死亡率效果不明显。 3.高压氧可能通过降低P53和Caspase-3的表达，从而对SAH后早期脑损伤起保护作用。
[Abstract]:Background and objective: subarachnoid hemorrhage (SAH) is one of the high mortality and high disability rate of cerebrovascular disease, which is divided into two types of spontaneous and traumatic. The proportion of SAH caused by intracranial aneurysm rupture is about 80% of all spontaneous SAH, also known as aneurysmal subarachnoid hemorrhage, its mortality and cause The mortality rate is very high, the death rate within 3 days after bleeding can reach 45%. scholar. The early brain injury after bleeding (early brain injury, EBI) is one of the main factors of high mortality and disability. The hyperbaric oxygen therapy (hyperbaric oxygentherapy, HBOT) is inhaling 100% oxygen in the atmosphere above a atmospheric pressure, and is now widely used in a variety of diseases. The treatment of the disease, due to the rapid change and risk of SAH patients, is very rare in the early treatment of SAH. This study established rat SAH animal model by intravascular puncture of internal carotid artery, observed the expression of P53 and Caspase-3 in the rat brain cortex, and the neurobehavioral, mortality and cerebral cortex of rats after hyperbaric oxygen drying. Changes in the expression of P53 and Caspase-3 in the mass.
Methods: 100 healthy adult male SD rats with a weight of 280-320g were randomly divided into 10 normal blank control group, 30 sham operation group (Group sham), 30 subarachnoid hemorrhage group (group SAH) and 30 SAH group (group HBOT+SAH) with hyperbaric oxygen intervention (group HBOT+SAH). 48h, 72h three subgroups, each subgroup 10. The blank control group did not do any special treatment, the routine feeding 3 days after anesthesia, perfusion, the brain tissue specimens preserved, SAH and HBOT+SAH group were modeled by internal carotid artery puncture method, after the modeling was successful, after the HBOT+SAH group rats were awakened after the anesthesia and hyperbaric oxygen treatment, the rest of the environment and the operation of the environment and the operation. The modeling process of the sham group was the same as that in the SAH group, but it did not break the blood vessel.Sham group, and the SAH and HBOT+SAH groups respectively recorded the neurobehavioral score and mortality of each subgroup of the subgroups after the successful modeling, 24h, 48h, 72h, and then anaesthetized and perfused the brain tissue of the subgroups of the rats. The Immunochemistry method was used to detect P in the cerebral cortex of rats. The expression of 53 and Caspase-3.
The Motic Image Advance3.2 image analysis system was used to analyze the immune tissue section and calculate the percentage of the positive cells. The Microsoft office Excel2003 software was recorded in the data. The normal distribution measurement data were expressed in S. The spss19.0 software was used to analyze the data, and the normal and variance homogeneity tests were carried out before the comparison, and the two groups were compared with t. The SNK method (q test) was used for comparison between the two subgroups. P < 0.05 had statistical significance.
Result锛

本文编号：1797246