笑氣對大鼠血腦屏障通透性的影響

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题名:	笑氣對大鼠血腦屏障通透性的影響 The Impact of Nitrous Oxide on Blood-Brain Barrier Permeability in Rats
作者:	徐仁國;Jen-Kuo Hsu
贡献者:	臨床醫學研究所碩士班
关键词:	血腦屏障;笑氣;blood brain barrier;nitrous oxide
日期:	2016-06-25
上传时间:	2017-03-22 14:21:03 (UTC+8)
出版者:	中國醫藥大學
摘要:	研究背景血腦屏障是維持中樞神經系統正常運作不可或缺的結構。特別是在腦部手術中，血腦屏障的完整與否關係到病人的預後。而手術中經常需要使用到吸入性麻醉藥，目前已知一些吸入性麻醉藥會對血腦屏障的通透性有所影響，此實驗的目的是要了解另一種吸入性麻醉藥-笑氣對於血腦屏障的影響。研究目的 Evans Blue dye是一種常用來評估血腦屏障對於大分子量物質通透性的試劑。此研究是利用尾靜脈注射Evans Blue dye到老鼠體內後，再用讓老鼠吸不同濃度的笑氣，測量大腦中不同區域的Evans Blue dye的含量，藉以評估笑氣是否會對血腦屏障造成影響。研究方法將老鼠分成五組(Control、20% N2O、40%N2O、80%N2O、20%Carbogen)，將老鼠用isoflurane麻醉，從尾靜脈打入Evans Blue dye，再將老鼠依實驗組別吸入不同濃度的氣體九十分鐘。九十分鐘後抽取股動脈的血液，再用生理食鹽水灌流十分鐘後，取出腦部不同區域的組織及肝臟組織。將血液、肝組織、腦組織用TCA淬取出內含的Evans Blue dye，再用spectrophotometry測量不同樣本的吸光值，經過換算後得到原始血液及組織內含的Evans Blue dye濃度，最後做統計分析。研究結果比較吸入不同濃度笑氣的老鼠，在不同腦區的Evans Blue dye含量並沒有顯著的差異性；與對照組(吸入空氣)的老鼠比較也沒有顯著的差異性。研究結論由此實驗可以證明從20%到80%的笑氣並不會改變大鼠在正常生理狀態下的血腦屏障對於Evans Blue dye的通透性。 BACKGROUND The blood-brain barrier plays an important role of maintaining the function of central nervous system. The integrity of blood-brain barrier is closely related to the prognosis of the patient especially in brain surgery. Inhalational anesthetics were frequently used during operation. Some inhalational anesthetics have variable degree of influence on blood-brain barrier permeability. The goal of this study is to uncover the impact on blood-brain barrier permeability of the nitrous oxide-an inhalational anesthetic. OBJECTIVE Evans Blue dye is an inert tracer frequently used to evaluate the permeability to high molecular weight substance of blood-brain barrier . After being injected with Evans Blue dye into the tail vein, the rats were then put into a translucent plastic box filled with different concentrations of nitrous oxide. We utilize spectrophotometry to detect the absorbance of residual Evans Blue dye in different brain regions to see if the nitrous oxide would alter the permeability of blood-brain barrier. METHODS The rats were allocated to five groups (Control、20%N2O、40%N2O、80%N2O、20%Carbogen). Isoflurane was used to anesthetize the rats. Evans Blue dye was injected into the tail vein of rats. Thereafter, the rats were put into a translucent plastic box filled with different concentrations of nitrous oxide according to its group for 90 minutes. Blood sample were collected from femoral artery after inhalation. After 10 minutes perfusion of normal saline, the brain was harvested for collecting samples from different brain region. A piece of liver tissue was also collected. Evans Blue dye contained in blood、liver and brain tissue was retrieved using trichloroacetic acid. Spectrophotometry was utilized to detect the absorbance of Evans Blue dye of samples. After converting the absorbance value to real content of Evans Blue dye of tissue, the content of Evans Blue dye of experimental groups was compared using statistics. RESULTS There was no significant difference of Evans Blue dye content of different brain regions among nitrous oxide groups, and there was also no significant difference when comparing the nitrous oxide groups and control group. CONCLUSIONS In conclusion, up to 80% nitrous oxide up don’t change the blood-brain barrier permeability to Evans Blue dye in rats under normal brain physiology.
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