【摘要】：腦出血破入腦室的高發(fā)病率、高死亡率和高致殘率給全球帶來沉重的經(jīng)濟負擔(dān)。然而迄今為止,針對腦出血外科手術(shù)治療及血壓管理的幾項臨床實驗均為陰性結(jié)果,1000余種神經(jīng)保護藥物的臨床轉(zhuǎn)化也均宣告失敗,腦出血臨床治療尚且缺乏有效的治療方法。目前基礎(chǔ)研究也多集中研究單純腦出血,針對腦出血破入腦室這一嚴重腦出血亞型的研究尚為一片空白,此外面對腦出血破入腦室后復(fù)雜的病理損傷機制,目前單一靶點、單一分子通路的神經(jīng)保護藥物研究恐難有成效。腦苷肌肽,一種復(fù)合型多靶點神經(jīng)保護藥物,具有高度血腦屏障通透性,其主要成分為多肽、多種神經(jīng)節(jié)苷脂、游離氨基酸、次黃嘌呤等,臨床應(yīng)用于治療中樞神經(jīng)及周圍神經(jīng)損傷疾病,基礎(chǔ)研究也已證實其在阿茲海默病、創(chuàng)傷性腦損傷、視網(wǎng)膜變性等多種動物模型中具有神經(jīng)保護作用,但其在出血性卒中的神經(jīng)保護效應(yīng)及機制目前仍不清楚。本實驗第一部分建立大鼠腦出血破入腦室模型,研究不同劑量腦苷肌肽對大鼠腦出血破入腦室后神經(jīng)功能恢復(fù)的影響;第二部分,研究腦苷肌肽在腦出血破入腦室后神經(jīng)保護效應(yīng)的可能作用機制和潛在靶點,為腦出血破入腦室的臨床治療提供新的思路及方法。一、Ⅳ型膠原酶誘導(dǎo)大鼠腦出血并破入腦室模型的建立及腦苷肌肽最適劑量探索目的通過對傳統(tǒng)大鼠腦出血模型進行改良,建立大鼠腦出血破入腦室標準化模型,運用行為學(xué)評估手段,評估多靶點復(fù)合神經(jīng)保護藥物腦苷肌肽不同劑量對大鼠腦出血后神經(jīng)功能恢復(fù)的影響。方法參照Rosenberg等提出的方法,并在本實驗室前期研究工作基礎(chǔ)上,調(diào)整Ⅳ型膠原酶注射坐標及劑量,將0.2U Ⅳ型膠原酶注入大鼠右側(cè)腦基底節(jié)區(qū)(前囟旁3毫米、后2毫米、深6毫米),3天后灌注取腦觀察血腫破入腦室情況。將大鼠隨機分為假手術(shù)組(Sham組),模型組(ICH組),模型+腦苷肌肽高劑量治療干預(yù)組(ICH+4ml/kg CEGI),模型+腦苷肌肽低劑量治療干預(yù)組(ICH+1ml/kg CEGI),模型+神經(jīng)節(jié)苷脂治療干預(yù)組(ICH+GM1),術(shù)后3天、7天、14天運用改良加西亞量表,轉(zhuǎn)角試驗兩種方法評估大鼠神經(jīng)功能恢復(fù)情況。結(jié)果1、術(shù)后3天可見血腫形成且破入側(cè)腦室,腦室擴張;2、神經(jīng)節(jié)苷脂與4ml/kg腦苷肌肽能夠顯著改善大鼠腦出血預(yù)后,促進大鼠神經(jīng)功能恢復(fù);3、低劑量腦苷肌肽干預(yù)組大鼠神經(jīng)功能缺損與模型組無明顯差異。結(jié)論此模型建立方法效果確切,成功模擬了臨床腦出血并破入腦室這一嚴重腦出血亞型,此外,行為學(xué)結(jié)果提示,4ml/kg腦苷肌肽能夠改善大鼠腦出血預(yù)后,其機制需要進一步探究。二、腦苷肌肽治療大鼠腦出血并破入腦室的機制研究目的在第一部分研究中,我們發(fā)現(xiàn)大鼠腦出血破入腦室在經(jīng)過4ml/kg腦苷肌肽連續(xù)干預(yù)14天后,其神經(jīng)功能缺損得到顯著改善,新近文獻表明腦苷肌肽的幾項主要組成成分具有抗氧化、促進軸突再生、再髓鞘化等作用,因此,本實驗擬探究腦苷肌肽治療大鼠腦出血破入腦室的可能機制。方法將實驗動物隨機分成假手術(shù)組(Sham組),模型組(ICH組),模型+腦苷肌肽治療干預(yù)組(ICH+4ml/kg CEGI)。術(shù)后2小時首次腹腔注射給藥,連續(xù)給藥14天。分別在術(shù)后3天、7天、14天運用小動物7.0T核磁共振成像動態(tài)觀察血腫、側(cè)腦室體積變化。術(shù)后7天、14天運用激光共聚焦、透射電子顯微鏡等技術(shù)手段動態(tài)觀察腦出血后血腫周圍及皮層白質(zhì)損傷。結(jié)果1、腦苷肌肽治療干預(yù)組術(shù)后第14天血腫吸收率較模型組顯著提高;2、腦苷肌肽治療干預(yù)組術(shù)后第14天側(cè)腦室體積擴張程度顯著減緩,遠期重度腦積水發(fā)生率顯著下降;3、腦苷肌肽治療干預(yù)組術(shù)后7天、14天皮層髓鞘堿性蛋白表達量顯著增加,血腫周圍白質(zhì)纖維束超微結(jié)構(gòu)損傷明顯改善。結(jié)論大鼠腦出血破入腦室,經(jīng)過腦苷肌肽干預(yù)后,出血后血腫周圍及皮層白質(zhì)損傷能夠顯著緩解,血腫吸收得到促進,側(cè)腦室體積擴張程度減緩,遠期重度腦積水發(fā)生率降低,神經(jīng)功能恢復(fù)得到促進,腦苷肌肽具有顯著的神經(jīng)保護作用,因此可作為腦出血破入腦室臨床治療的新方法。
[Abstract]:The high morbidity, high mortality and high disability rate of the cerebral hemorrhage to the ventricles have brought a heavy economic burden to the world. So far, the clinical conversion of more than 1000 kinds of neuroprotective drugs has failed due to the negative result of several clinical trials for the surgical treatment of cerebral hemorrhage and the management of blood pressure, and the clinical treatment of the cerebral hemorrhage is still short and the effective treatment method is not available. The present basic research also focuses on the study of the simple cerebral hemorrhage, and the study on the cerebral hemorrhage of the cerebral hemorrhage is a blank, and in addition to the complicated pathological damage mechanism after the cerebral hemorrhage is broken into the ventricle, the single target point is not present at present. The study of the neuroprotective drug of a single molecular pathway may be difficult to achieve. the brain papillary muscle peptide, a compound multi-target nerve protection medicine, has the high blood-brain barrier permeability, the main component of the brain-derived muscle peptide is a polypeptide, a plurality of ganglionic fat, a free amino acid, a secondary flavopterin and the like, and is clinically applied to the treatment of the central nervous system and the peripheral nerve injury disease, The underlying studies have also been shown to have neuroprotective effects in various animal models, such as Alzheimer's disease, traumatic brain injury, and retinal degeneration, but their neuroprotective effects and mechanisms in hemorrhagic stroke are still unclear. In the first part of this experiment, the cerebral hemorrhage of rats was established to break into the ventricular model, and the effect of different doses of the brain myostatin on the recovery of the neurological function after the cerebral hemorrhage of the rat was broken into the ventricle was studied. The second part, To study the possible mechanism and potential target of the neuroprotective effect of the brain-derived myopeptide in the post-cerebral hemorrhage of the cerebral hemorrhage, and to provide a new thought and method for the clinical treatment of the cerebral hemorrhage. one, type 鈪，

本文編號：2388818