急性高血糖对大鼠脑出血后神经损伤的影响

发布时间：2018-05-08 21:30

  本文选题：脑出血 + 高血糖　； 参考：《浙江大学》2013年硕士论文

【摘要】：背景与目的： 脑出血(intracerebral hemorrhage,ICH)是神经系统的常见疾病,占急性脑血管病的10%-15%,死亡率与致残率极高。脑出血后除血肿的占位效应导致原发性损伤外,继发性损伤如血肿周围脑组织发生一系列病理变化也是导致脑出血患者神经损伤加重、预后不良的重要因素。近年来发现许多脑出血(ICH)患者入院时存在高血糖且预后不良,本实验建立伴有急性高血糖状态的大鼠脑出血模型来研究急性高血糖对大鼠脑出血后神经损伤的影响。 长期以来自噬被认为是细胞的一种重要分解代谢途径,可清除细胞内衰老细胞器和错误折叠蛋白,在缺氧缺血等应激状态下有助于维持细胞能量稳态、促进细胞存活。近年来研究发现自噬参与多种疾病的病理生理过程,ICH后自噬也激活,但自噬激活后究竟是加重ICH后神经损伤还是作为神经功能保护机制至今仍未明确。细胞自噬水平与细胞本身生理病理状态及周围环境等密切相关,微管相关蛋白轻链3(LC3)和Beclin-1是两个经典的自噬标记物,本实验通过检测血肿周围细胞LC3和Beclin-1水平来观察自噬变化并研究ICH后高糖对自噬的影响。 研究方法： 成年SD雄性大鼠随机分成四组,即正常对照组(C组)、高血糖组(G组)、脑出血组(Ⅰ组)、伴急性高血糖的脑出血组(IG组)。大鼠右侧尾状核注射股动脉自体血建立脑出血动物实验模型(Ⅰ组),注血前30分钟腹腔注射50%葡萄糖溶液(6ml/kg)建立伴有急性高血糖的大鼠脑出血模型(IG组)。观察术前及术后24小时神经功能缺失症状(转角试验及前肢运动不对称试验)并评分、术后24小时血肿大小、脑组织含水量及脑组织HE染色切片,评估ICH后神经损伤程度；免疫组化及免疫印迹技术(Western blot, WB)检测术后24小时血肿周围脑组织Beclin-1和LC3蛋白,评估血肿周围细胞自噬水平变化。 结果： 1.脑出血24小时后转角试验及前肢运动不对称试验评分均升高(P0.01),血肿周围脑组织含水量升高(P0.05),组织切片HE染色可见血肿周围组织水肿明显,细胞分布不均匀,神经元细胞数量明显减少,中性粒细胞及淋巴细胞浸润。 2.脑出血24小时后脑组织LC3和Beclin-1蛋白免疫组化染色血肿周围可见阳性细胞,较正常对照组染色深,WB结果可见血肿周围组织LC3和Beclin-1蛋白水平升高。 3.无ICH的单纯高血糖不影响大鼠神经功能缺失症状评分及脑含水量,但与血糖正常的脑出血组相比,急性高血糖可使脑出血24小时后前肢运动不对称试验评分升高(P0.01),血肿周围脑组织含水量升高(P0.01)。 4.无ICH的单纯高血糖不影响大鼠脑右侧基底节LC3和Beclin-1蛋白水平,但与血糖正常的脑出血组相比,急性高血糖使脑出血24小时后血肿周围LC3和Beclin-1阳性细胞染色变浅,并降低血肿周围组织LC3和Beclin-1水平。 结论： 1.大鼠脑出血后出现明显神经功能缺失症状和脑水肿,血肿周围组织自噬水平升高。 2.急性高血糖可加重大鼠脑出血后神经功能缺失症状和脑水肿,降低血肿周围细胞自噬水平。
[Abstract]:Background and purpose:
Intracerebral hemorrhage (ICH) is a common disease of the nervous system, which accounts for the 10%-15% of acute cerebrovascular disease. The mortality and disability rate are very high. The secondary injury, such as the pathological changes of the brain tissue around hematoma, is also the cause of nerve injury in the patients with cerebral hemorrhage. In recent years, many patients with cerebral hemorrhage (ICH) have high blood sugar and poor prognosis. In this experiment, a rat model of cerebral hemorrhage with acute hyperglycemia was established to study the effect of acute hyperglycemia on the nerve injury after cerebral hemorrhage in rats.
Autophagy has long been considered to be an important catabolic pathway for cells, which can remove intracellular senescent organelles and misfolded proteins. In the stress state of hypoxia and ischemia, it helps to maintain cell energy homeostasis and promote cell survival. In recent years, autophagy has been found to be involved in the pathophysiological process of multiple diseases. Autophagy is also activated after ICH. But it is still not clear whether the autophagy is aggravated after the activation of ICH or as a protective mechanism for nerve function. The level of autophagy is closely related to the physiological and pathological state of the cells and the surrounding environment. Microtubule related protein light chain 3 (LC3) and Beclin-1 are two classic autophagic markers. This experiment is done by detecting the hematoma around the hematoma. Cell LC3 and Beclin-1 levels were used to observe the changes of autophagy and the effect of high glucose after ICH on autophagy.
Research methods:
Adult SD male rats were randomly divided into four groups, namely, normal control group (group C), hyperglycemia group (group G), cerebral hemorrhage group (group I), group of cerebral hemorrhage (group IG) with acute hyperglycemia (group IG). Rat right caudate nucleus injection of femoral artery autologous blood to establish an animal experimental model of cerebral hemorrhage (group I), intraperitoneal injection of 50% glucose solution (6ml/kg) before injection of blood (6ml/kg) was established. Acute hyperglycemic rat model of cerebral hemorrhage (group IG). The symptoms of nerve function loss (angle test and forelimb motion asymmetry test) 24 hours before and after operation were observed, and the size of hematoma, water content of brain tissue and HE staining section of brain tissue were observed after 24 hours of operation, and the degree of deity injury after ICH was evaluated; immunoblotting and immunoblotting (Wester N blot (WB) was used to detect the Beclin-1 and LC3 proteins around the hematoma around 24 hours after operation, and to assess the changes of autophagy level around the hematoma.
Result锛，

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