本文選題：腦出血 + 高血糖�。� 參考：《浙江大學(xué)》2013年碩士論文

【摘要】：背景與目的： 腦出血(intracerebral hemorrhage,ICH)是神經(jīng)系統(tǒng)的常見疾病,占急性腦血管病的10%-15%,死亡率與致殘率極高。腦出血后除血腫的占位效應(yīng)導(dǎo)致原發(fā)性損傷外,繼發(fā)性損傷如血腫周圍腦組織發(fā)生一系列病理變化也是導(dǎo)致腦出血患者神經(jīng)損傷加重、預(yù)后不良的重要因素。近年來發(fā)現(xiàn)許多腦出血(ICH)患者入院時(shí)存在高血糖且預(yù)后不良,本實(shí)驗(yàn)建立伴有急性高血糖狀態(tài)的大鼠腦出血模型來研究急性高血糖對大鼠腦出血后神經(jīng)損傷的影響。 長期以來自噬被認(rèn)為是細(xì)胞的一種重要分解代謝途徑,可清除細(xì)胞內(nèi)衰老細(xì)胞器和錯(cuò)誤折疊蛋白,在缺氧缺血等應(yīng)激狀態(tài)下有助于維持細(xì)胞能量穩(wěn)態(tài)、促進(jìn)細(xì)胞存活。近年來研究發(fā)現(xiàn)自噬參與多種疾病的病理生理過程,ICH后自噬也激活,但自噬激活后究竟是加重ICH后神經(jīng)損傷還是作為神經(jīng)功能保護(hù)機(jī)制至今仍未明確。細(xì)胞自噬水平與細(xì)胞本身生理病理狀態(tài)及周圍環(huán)境等密切相關(guān),微管相關(guān)蛋白輕鏈3(LC3)和Beclin-1是兩個(gè)經(jīng)典的自噬標(biāo)記物,本實(shí)驗(yàn)通過檢測血腫周圍細(xì)胞LC3和Beclin-1水平來觀察自噬變化并研究ICH后高糖對自噬的影響。 研究方法： 成年SD雄性大鼠隨機(jī)分成四組,即正常對照組(C組)、高血糖組(G組)、腦出血組(Ⅰ組)、伴急性高血糖的腦出血組(IG組)。大鼠右側(cè)尾狀核注射股動脈自體血建立腦出血動物實(shí)驗(yàn)?zāi)Ｐ?Ⅰ組),注血前30分鐘腹腔注射50%葡萄糖溶液(6ml/kg)建立伴有急性高血糖的大鼠腦出血模型(IG組)。觀察術(shù)前及術(shù)后24小時(shí)神經(jīng)功能缺失癥狀(轉(zhuǎn)角試驗(yàn)及前肢運(yùn)動不對稱試驗(yàn))并評分、術(shù)后24小時(shí)血腫大小、腦組織含水量及腦組織HE染色切片,評估ICH后神經(jīng)損傷程度；免疫組化及免疫印跡技術(shù)(Western blot, WB)檢測術(shù)后24小時(shí)血腫周圍腦組織Beclin-1和LC3蛋白,評估血腫周圍細(xì)胞自噬水平變化。 結(jié)果： 1.腦出血24小時(shí)后轉(zhuǎn)角試驗(yàn)及前肢運(yùn)動不對稱試驗(yàn)評分均升高(P0.01),血腫周圍腦組織含水量升高(P0.05),組織切片HE染色可見血腫周圍組織水腫明顯,細(xì)胞分布不均勻,神經(jīng)元細(xì)胞數(shù)量明顯減少,中性粒細(xì)胞及淋巴細(xì)胞浸潤。 2.腦出血24小時(shí)后腦組織LC3和Beclin-1蛋白免疫組化染色血腫周圍可見陽性細(xì)胞,較正常對照組染色深,WB結(jié)果可見血腫周圍組織LC3和Beclin-1蛋白水平升高。 3.無ICH的單純高血糖不影響大鼠神經(jīng)功能缺失癥狀評分及腦含水量,但與血糖正常的腦出血組相比,急性高血糖可使腦出血24小時(shí)后前肢運(yùn)動不對稱試驗(yàn)評分升高(P0.01),血腫周圍腦組織含水量升高(P0.01)。 4.無ICH的單純高血糖不影響大鼠腦右側(cè)基底節(jié)LC3和Beclin-1蛋白水平,但與血糖正常的腦出血組相比,急性高血糖使腦出血24小時(shí)后血腫周圍LC3和Beclin-1陽性細(xì)胞染色變淺,并降低血腫周圍組織LC3和Beclin-1水平。 結(jié)論： 1.大鼠腦出血后出現(xiàn)明顯神經(jīng)功能缺失癥狀和腦水腫,血腫周圍組織自噬水平升高。 2.急性高血糖可加重大鼠腦出血后神經(jīng)功能缺失癥狀和腦水腫,降低血腫周圍細(xì)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錛，

本文編號：1863136