丹參與槐花對大鼠腦缺血-再灌流損傷引發之腦梗塞的效用; Effects of Salvia miltiorrhiza and Sophora japonica on Cerebral Infarct Induced by Ischemia-Reperfusion Injury Rats

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Title:	丹參與槐花對大鼠腦缺血-再灌流損傷引發之腦梗塞的效用;Effects of Salvia miltiorrhiza and Sophora japonica on Cerebral Infarct Induced by Ischemia-Reperfusion Injury Rats
Authors:	姥芝瑞;Lao chih jui
Contributors:	中國醫藥大學中國醫學研究所
Keywords:	丹參;槐花;腦梗塞;缺血再灌流;自由基;細胞凋亡;Salvia Miltorrhiza;Sophora Japonica;Cerebral Infarct;Ischemia-reperfusion;Free radicals;ED1;interleukin-1β;Apoptosis.
Date:	2004
Issue Date:	2010-01-20
Abstract:	傳統中醫學認為血瘀為腦中風的主要因素之一，因此臨床上常使用活血化瘀的中藥來治療。丹參(Salvia miltiorrhiza Bge)、槐花(Sophora japonica L)為傳統的中藥。丹參具有活血化瘀的功效，臨床上常用來治療腦中風和缺血性心臟疾病，一些研究報告說明在血瘀證的兔子或老鼠，丹參具有改善微循環、血液黏稠度和紅血球的變形性的能力。槐花具有涼血、止血、清肝瀉火的作用，臨床上常用來治療痔瘡出血。最近的研究發現，槐花與銀杏都含有相同的成份Rutin和Quercetin，銀杏已被廣泛的應用在心血管疾病和腦痴呆的治療。本研究的目的是探討丹參與槐花對腦梗塞的效用，並探討其可能作用機轉。方法是將300-350公克的Sprague-Dawley(SD)大鼠的兩側總頸動脈和右側中大腦動脈的腦血流阻斷90分鐘，經24小時的再灌流，建立一個大鼠腦缺血-再灌流損傷引發之腦梗塞之動物模型。總共使用84 隻Sprague-Dawley大鼠，丹參組30隻，槐花組54隻。丹參組分別使用丹參15 mg/kg、30 mg/kg，而槐花組分別使用槐花50 mg/kg、100 mg/kg、200 mg/kg於腦血流阻斷前30分鐘，以及槐花200 mg/kg於腦血流阻斷後30分鐘，從腹腔注射給藥。經24小時再灌流後，將動物犧牲取腦並作成切片，經2 % 2,3,5-triphenyl-terazolium chloride染色後，計算腦梗塞面積與切片總面積之百分比。此外，丹參組同時測量周邊血液中的Luminol-chemiluminesenence (CL) counts和Lucigenin - CL counts。槐花組觀察大鼠神經缺損等級及計算腦梗塞區域的ED1、interleukin-1β(IL-1 β)染色陽性細胞和apoptosis細胞。以大鼠腦梗塞面積百分比的變化來評估丹參對腦梗塞的效用；以神經缺損等級、腦梗塞面積百分比的變化，來評估槐花對腦梗塞的效用。結果顯示丹參15 mg/kg 、30 mg/kg於腦血流阻斷前30分鐘給藥能減少梗塞面積和Luminol-CL counts。槐花100、200 mg/kg於腦血流阻斷前30分鐘給藥以及槐花200 mg/kg於腦血流阻斷後30分鐘給藥都能減少腦梗塞面積和神經缺損狀態。另外，槐花200 mg/kg於腦血流阻斷前30分鐘給藥能減少腦梗塞區的ED1、IL-1β染色陽性細胞和apoptosis細胞的數目。總結，丹參能減少大鼠的腦梗塞面積和Luminol-CL count。槐花能減少大鼠腦梗塞和神經缺損狀態，推測丹參與槐花可應用於治療人類的腦梗塞。丹參減少腦梗塞面積的作用部分與自由基的清除有關，而槐花減少腦梗塞面積的作用與抑制發炎細胞microglia、IL-1β以及apoptosis有關。; According to the theory of traditional Chinese medicine, blood stasis could result in cerebral infarction. It could be improved by the medicine that could quicken the blood and dispell stasis. Both Salvia Miltorrhiza Bunge (SM) and Sophora Japonica L.(SJ) are traditional Chinese herbs. Since Salvia Miltorrhiza Bunge quickens the blood and dispells stasis, it is commonly used to treat disorders of blood stasis such as ischemic brain (cerebral infarct) and heart diseases. Because Sophora Japonica L. cools the blood and stops bleeding, it is used to treat hemorrhoid with bleeding. Both the SJ and the Ginkgo biloba have components of quercetin and rutin. Ginkgo biloba has been widely used to treat cerebrovascular disorders and dementia in humans. The aim of the present study was to investigate the effect of SM and SJ on cerebral infarct. A total of 30 Sprague-Dawley (SD) male rats were used in the SM study and 54 in the SJ study. Focal cerebral infract was established by occluding both the bilateral common corotid arteries and the right middle cerebral artery for 90 minutes. SM and SJ were administered intraperitoneally 30 minutes prior to blocking the blood flow, respectively. In addition, SJ was administered intraperitoneally 30 minutes after the blood flow occlusion. Twenty-four hrs after reperfusion, the rats were killed and their brain tissue was stained with 2,3,5-triphenly-terazolium chloride and the areas of infarct were calculated. The luminal - chemiluminesence (CL) counts and lucigenin-CL counts of peripherial blood were measured in the SM study. The neurological status, ED1,interleukin-1β(IL-1β)- immunoreacting cells, and apoptotic cells were also measured in the cerebral infarct zone in the SJ study. Pretreatment with the intraperitoneal injection of 30 mg/kg and 15 mg/kg of SM reduced the area of cerebral infarct and the luminol-CL counts of peripheral blood. In addition, pretreatment with SJ at 100 mg/kg, 200 mg/kg and post-treatment with SJ at 200 mg/kg reduced the area of cerebral infarct by % and the grading scale of neurological deficit. Pretreatment with SJ at 200 mg/kg also reduced ED1, interleukin-1β- immunoreacting cells, and apoptotic cells. These results suggest that the SM and the SJ are potential drugs to protect against the cerebral infarct in humans. The neuroprotective effect of the SM may be partially due to free radical scavenging activities. As for the SJ, its suppressive action of microglia, IL-1β and apoptosis could result in the improvement of the cerebral infract.
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