大鼠脑出血后线粒体DNA水平下降与平肝熄风汤的作用

Effect of pinggan xifeng decoction on decreasing the level of mitochondrial DNA in rats with intracerebral hemorrhage

目的:观察大鼠脑出血后线粒体DNA(mtDNA)水平的改变,及中药平肝熄风汤的干预作用。方法:实验于2003-11在湘雅医院中心实验室进行,取SD大鼠65只,随机分为4组:正常对照组(n=5),假手术组、模型组和平肝熄风汤组(n=20);后3组于造模后设4h,1,3,7d4个时间点,每个时间点5只。①造模:模型组和平肝熄风汤组用0.4UⅦ型胶原酶于脑内定位注射诱导大鼠脑出血模型,假手术组注入灭菌生理盐水2μL,正常对照组不干预。②给药:平肝熄风汤组动物清醒后开始灌服平肝熄风汤穴钩藤15g,大黄10g,天竺黄5g,三七6g等雪生药82.52g/穴kg·d雪,每天上下午各1次,3mL/次。其他3组给予灭菌生理盐水灌服。③在相应时间点测定神经缺损症状积分(从向出血侧旋转,前爪抓力,爬竿能力3个方面评定,每项0～5分,0分为正常),然后处死,运用紫外分光光度计测定脑海马mtDNA水平。结果:65只大鼠进入结果分析。①神经缺损症状积分:模型组和平肝熄风汤组大鼠于造模后4h达高峰(8.90±1.14,8.80±0.84),随后逐渐下降,至7d仍显著高于假手术组和正常组穴P<0.01雪;平肝熄风汤组1,3,7d均低于模型组穴P<0.05,0.01雪。②mtDNA水平:模型组和平肝熄风汤组各时间点均显著低于假手术组(P<0.01),假手术组与正常对照组无差别(P>0.05),平肝熄风汤组造模后3,7d两个时间点明显高于模型组(P<0.05)。结论:大鼠脑出血后mtDNA水平下降,平肝熄风汤能阻断mtDNA水平的下降,而发挥其神经保护作用。

AIM： To observe the interventional effect of pinggan xifeng decoction （PGXFD） on the change of mitochondrial DNA （mtDNA） content in rats with intracerebral hemorrhage （ICH） injury. METHODS： The experiment was completed at Central Laboratory of Xi- angya Hospital in November 2003. Totally 65 SD rats were randomly divided into four groups： normal control group （n=5）, sham operation group （n=20）, model group （n=20） and PGXFD group （n=20）. Four time points which were 4 hours, 1, 3 and 7 days were set after operation with 5 rats in each group after modeling ①Modeling： 0.4 U type Ⅶ collagenase was stereotaxically injected into the brains of rats in model group and PGXFD groups to establish intracerebral hemorrhage model; 2 μL saline was injected into the rats in sham operation group; rats in normal control group were not treated. ② Administration： 82.52 g/（kg ·d） raw PGXFD was perfused into rats in PGXFD group after wakefulness for once every morning and afternoon with 3 mL a time. PGXFD was consisted of gouteng 15 g, dahuang 10 g, tianzhuhuang 5 g, sanqi 6 g, etc. Rats in other three groups were perfused with sterile saline. ③ Scores of neurological defect were measured in the corresponding time points on the aspects of turning to hemorrhage side, grasping with anterior claw, and climbing ability （each item scores from 0 to 5 points, and 0 point was regarded as normality）, then rats were sacrificed. Level of hippocampal mtDNA was detected with ultraviolet spectrophotometer. RESULTS： Totally 65 rats entered the final analysis. ① Scores of neurological defects of rats in model group and PGXFD group reached the peak 4 hours after modeling （8.90±1.14, 8.80±0.84）, and then decreased gradually, but were still more than those in sham operation group and normal group on the 7^th day （P 〈 0.01）. Scores in PGXFD group were less than those in model group on the 1^st, 3^rd and 7^th days （P 〈 0.05, 0.01）. ② mtDNA level in model group and PGXFD group was lower than that in sham operation group at each time point （P 〈 0.01）, and there was not significance between sham operation group and normal control group （P 〉 0.05）. mtDNA level in PGXFD group was higher than that in model group at 3^rd and 7^th days after modeling （P 〈 0.05）. CONCLUSION： mtDNA content decreases in rats suffered from ICH. PGXFD can protect neuron through inhibiting the decrease of mtDNA content.