探討三黃瀉心湯與黃芩素在大鼠蜘蛛膜下腔出血模式的治療效果

Effects of San-Huang-Xie-Xin-Tang and Baicalein on Subarachnoid Hemorrhage in Rat

許博淳(Po-Chun Hsu)

指導教授 ： 關皚麗

共同指導教授 ： 葉竹來(Jwu-Lai Yeh)

高雄醫學大學/醫學院/藥理學科研究所/碩士(2010年)

摘要

蜘蛛膜下腔出血（SAH）是一種嚴重且致命的疾病，但是這項疾病的致病機轉仍然不是非常清楚。在先前的研究結果顯示，在蜘蛛膜下腔出血後急性的腦部局部出血會壓迫到既有的血液循環，導致全腦的缺血，繼而引發進一步的蛋白與細胞激素的釋放，最終導致腦神經元的死亡。在神經元死亡之後會再引發腦部水腫或是血腦障壁的缺損情形，最終引發腦血管痙攣現象。
先前的研究指出三黃瀉心湯具有胃保護、治療肺高壓、減少心肌梗塞等作用。然而三黃瀉心湯或是其中所含有的活性成分都尚未在蜘蛛膜下腔出血模式中被研究，因此本實驗的目的為探討三黃瀉心湯在大鼠蜘蛛膜下腔出血下的預防與治療作用，評估兩種不同投藥方式下的治療效果，並且選擇黃芩素（Baicalein）作為效果對照。
實驗利用大鼠蜘蛛膜下腔出血二次模式進行，在第零與二天進行自體腦池注射尾動脈血液，以模擬蜘蛛膜下腔出血病程發展。最後在第七天時將大鼠犧牲。期間大鼠每日的體重變化都會被記錄以觀察活動力與食慾的變化，操作蜘蛛膜下腔出血時得到的腦脊髓液也會利用明膠蛋白酵素電泳法（Gelatin Zymography）分析腦脊髓液中的基質金屬蛋白酶活性。腦基底動脈會取下後觀察腦血管痙攣的情形，海馬迴部分也會使用西式點墨法評估蛋白表現的變化。
三黃瀉心湯能夠抑制轉化生長因子的釋放，以及MMP的活性與表現；也能夠抑制MAPK的磷酸化，進而抑制下游的COX-2、Bax表現，進而抑制腦部神經元細胞的凋亡以及發炎反應。三黃瀉心湯也能夠抑制HIF-1α的生成，進而使得ET-1的量減少，達到抑制血管痙攣的效果。總體而言，三黃瀉心湯的優點在於成分複方，不同的有效成分間可達到協同的治療作用，且傳統中藥取得較為方便且經濟；黃芩素的優點則在於處方單一，可個別分析各種標的上的治療效果，成分也較容易控制，也能促進大鼠的活力與食慾。

關鍵字

黃芩素 ； 蜘蛛膜下腔出血 ； 三黃瀉心湯

並列摘要

Subarachnoid hemorrhage (SAH) ranks a lethal disease worldwide. Its pathology, mechanism and treatment remains not fully understood. Previous studies showed that blood would suppress the circulation of whole brain, causing cerebral ischemia. Cerebral ischemia caused cytokines released and cell signal transduction. The results are the death of neuron, and caused brain edema, Blood-brain barrier (BBB) disruption and vasospasm.
San-Huang-Xie-Xin-Tang (SHXT) has been used to gastric protection, pulmonary hypertension and myocardial infarction. The effects of SHXT and its active components have not been discussed. In this study, we will investigate the neuroprotective effects of SHXT in a reproducible rodent SAH model, and baicalein was used for effects comparing.
Male SD rats were anesthetized and blood was drawn from tail artery and injected into the cisterna magna twice separated by 2 days to induce SAH. Rats were separated into 5 groups. There was (1) Normal, (2) SAH, (3) Prevention of SHXT, (4) Treatment of SHXT and (5) Baicalein groups. Rat weight was recorded for assessment of appetite and activity. The basilar artery was then obtained and embedded for further histological examination. Cerebrospinal fluid was obtained for gelatin zymography to measure the MMPs activity. The CA1 area of hippocampus tissue was homogenized for western blotting analysis.
SHXT and baicalein decreased brain inflammation and neuron apoptosis related proteins in hippocampus. The results offered evidence that SHXT and baicalein attenuate brain inflammatory, ET-1, TGF-β, HIF-1α, MAPK, cytokine expression and neuron apoptosis. Further, the neuroprotective effect of SHXT and baicalein could be investigated for further use.

並列關鍵字

baicalein ； SAH ； SHXT ； San-Huang-Xie-Xin-Tang ； Subarachnoid Hemorrhage

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