涤痰汤对脑出血大鼠神经元线粒体内细胞色素C释放的影响  

Effects of ditan decoction on the release of Cytochrome C from neuronal mitochondria in rats with intracerebral hemorrhage

作　　者：王立[1] 梁清华[1] 王涛[2] 包太成[1] 侯俊良[1] 

机构地区：[1]中南大学湘雅医学院中西医结合研究所,湖南省长沙市410008 [2]中南大学湘雅二医院眼科,湖南省长沙市410011

出　　处：《中国临床康复》2005年第25期61-63,i0005,共4页Chinese Journal of Clinical Rehabilitation

基　　金：湖南省自然科学基金(01JJY2024);湖南省科技基金(1013-50)~~

摘　　要：目的:观察大鼠脑出血后血肿周围神经元线粒体内激活半胱氨酸天冬氨酸蛋白酶及诱导细胞凋亡的细胞色素C的释放规律以及血肿变化和中药涤痰汤的干预作用。方法:实验于2003-10/2004-03在中南大学中西医结合研究所实验室进行。取健康SD大鼠65只,随机分成4组:正常组(n=5),假手术组(n=20),模型组(n=20),涤痰汤组(n=20),除正常组外,其余3组设4个时间点:6h,1,3,5d,每个时间点5只大鼠。正常组不干预,模型组和涤痰汤组用脑内立体定位注射含0.4UⅫ型胶原酶的生理盐水2μL诱导大鼠脑出血模型,假手术组注入2μL生理盐水。涤痰汤组大鼠手术清醒后即灌服涤痰汤中药煎液(半夏、南星各12g,橘红7.5g,枳实、赤茯苓各10g,石菖蒲、人参各5g,竹茹3.5g,甘草2.5g)生药2.67g/(kg·d),模型组和假手术组用蒸馏水灌胃,2mL/次,2次/d。神经元线粒体内细胞色素C的释放情况应用免疫组织化学法检测,不同时间点大鼠脑内血肿的最大直径用直尺测量。结果:65只动物均进入结果分析。①脑组织细胞色素C阳性细胞计数:正常组未见阳性细胞;假手术组仅于6h,1d时见少数阳性表达细胞,余时间点未见阳性细胞;模型组血肿周围于6h出现阳性表达细胞,1d阳性细胞计数达高峰,3d出现轻度下降,5d明显下降犤(12.66±2.12),(51.74±2.13),(35.46±2.84),(17.34±2.08)个犦;涤痰汤组与同时间模型组比较在6h计数无明显差异犤(12.68±2.06)个,P>0.05犦,1,3,5d时阳性细胞减少明显犤(50.28±2.87),(31.12±3.05),(13.18±2.94)个,P<0.05犦。②脑血肿的最大直径:正常组和假手术组无血肿形成;模型组造模后于1d达到最大,3d有所变小,5d最小犤(4.08±0.16),(3.58±0.15),(2.48±0.15)mm犦;涤痰汤组在3d时血肿小于模型组犤(3.22±0.26)mm,P<0.05犦,5d时差距更明显犤(1.70±0.19),P<0.01犦。结论:脑出血后血肿周围的神经元细胞色素C释放明显增多,而且是脑出血的一个早期事件,从脑出血�AIM： To observe the release rule of the activation of cysteine aspartyl proteinase and the inductive apoptosis of Cytochrome C （Cyt-C） in neuronal mitochondrion around hematom after intraeerebral hemorrhage （ICH） in rats and the changes of hematoma and the internention effect of the traditional Chinese medicine, ditan decoction （DTT）. METHODS： The experiment was carried out in the Institute of Combined Traditional Chinese and Western Medicine of Central South University from October 2003 to March 2004, Sixty-five healthy SD rats were selected and divided randomly into four groups： the normal group （n=5）, the sham operation group （n=20） , the model group （n=20）, the ditan decoction group （n=20）, four time points （6 hours, 1 day, 3 days, and 5 days） was made except the normal group, five in every time point. Without interfere in the normal group. 2 μL saline with 0.4 U VⅡ type collagenase was stereotaxi-cally injected into the brain in rats to induce the ICH model in the model group and the ditan decoction group, and 2 μL saline was injected into the rats in sham operation group. The DTT traditional Chinese medicine liquid（pinellia tuber and southern star with 12g, respectively, red tangerine peel 7.5g, immature bitter orange and red India bread with 10g, respectively, grassleaved sweetflag rhizome and ginseng with 5g, respectively, bamboo shavings 3.5g and liquorice root 2.5 g） crude drug 2.67g/kg per day was drenched in awake rats after the operation in ditan decoction group. The rats in the model group and the sham operation group were treated with gastric perfusion of distilled water with 2mL every time and twice a day. Immunohistochemieal method was used to exam the release of Cyt-C from neuronal mitochondria and a ruler was used to measure the maximum diameter of hematoma in rat＇s brain at different time points. RESULTS： Sixty-five rats were all involved in the result analysis. ① The positive cells quantity of Cyt-C in brain tissue： The positive cells did

关 键 词：脑出血/中药疗法 细胞色素C 大鼠 

分 类 号：R745[医药卫生—神经病学与精神病学]