大黄苷元联合溶栓对血栓栓塞性脑缺血大鼠肺胃损伤的保护作用

Protective effects of Rhubarb Aglycone combined with thrombolysis on the injury of lung and stomach in rats with thrombus-occluded cerebral ischemia

目的:观察大黄苷元联合溶栓治疗对大鼠脑缺血损伤肺胃组织的保护作用。方法:实验于2005-08/2006-07在河南中医学院老年医学研究所实验室完成。①260只SD大鼠采用随机数字法分为假手术组20只、模型组60只、尿激酶组60只、大黄苷元组60只、大黄苷元+尿激酶组60只;除假手术组外,其余各组根据缺血后动脉用药时间又各分为3,6,9h3个时间点,每个时间点20只。②自体血栓结合线栓阻塞大鼠大脑中动脉制备局灶性脑缺血动物模型。③各组大鼠均于术前4d灌胃用药,大黄苷元组、大黄苷元+尿激酶组用大黄苷元灌胃(灌胃体积为每100g大鼠1mL),假手术组、模型组和尿激酶组用等体积的生理盐水灌胃;动脉用药除假手术组外,其余各组分别于造模后3,6,9h经导管由区域动脉给药,尿激酶组与大黄苷元+尿激酶组用尿激酶(用药体积为20μL),模型组和大黄苷元组区域动脉用同等体积的生理盐水。④动脉给药后24h,观察大鼠脑组织病理损伤、颅内和胃出血率、脑和肺组织含水量、肺和胃病理损伤变化。结果:实验过程中因麻醉、操作等原因死亡及剔除大鼠156只,进入结果分析104只。①颅内和胃出血率:尿激酶组9h大鼠颅内出血率较模型组高(66.67%,28.57%,P<0.05);尿激酶组9h脑和胃出血率较3h高(脑:66.67%,18.75%;胃:41.18%,17.65,P<0.05);大黄苷元+尿激酶组9h颅内出血率较尿激酶组9h低(P<0.05)。②脑和肺及胃组织病理改变:各模型组大鼠脑、胃和肺组织病理损伤均较假手术组明显;各用药组脑和肺组织分别较相应时间模型组减轻;各组脑、胃和肺组织损伤9h均较其3h明显;大黄苷元+尿激酶组9h较相应时间点尿激酶组和大黄苷元组损伤减轻(P<0.05)。③脑和肺组织含水量:各模型组脑和肺组织含水量均较假手术组增高(P<0.01);尿激酶组和大黄苷元+尿激酶组各时间点均较模型组降低(P<0.01);各组9h分别较其3h脑和肺含水量增加(P<0.01,P<0.05);大黄苷元+尿激酶组6h脑组织和9h肺含水量分别较尿激酶组降低(P<0.05)。结论:脑缺血后延迟溶栓治疗可引起大鼠脑和胃出血率增高、脑组织和肺组织水肿加重,脑和肺及胃组织病理损伤明显;大黄苷元联合溶栓可降低脑出血率,改善神经细胞超微结构,降低脑和肺组织含水量,对脑缺血肺和胃组织损伤具有保护作用。

AIM： To observe protective effects of Rhubarb Aglycone combined with thrombolysis therapy on injury of lung and stomach in rats with cerebral ischemia. METHODS： The experiment was performed from August 2005 to July 2006 in Laboratory of Institute of Gerontological Medicine, Henan University of Traditional Chinese Medicine. （1）260 SD rats were randomly divided into sham operation group （n =20）, model group （n =60）, urokinase group （n =60）, Rhubarb Aglycone group （n =60）, Rhubarb Aglycone plus urokinase group （n =60）. Other groups were divided into 3-hour, 6-hour and 9-hour groups again respectively according to administration time through artery of postischemia except sham operation group, 20 rats in each subgroup. （2） Thrombus-occluded cerebral ischemia model group was duplicated by autologous blood blot and inserted nylon thread. （3）Rats in each group were administrated through gastric perfusion 4 days before operation. Rats in Rhubarb Aglycone group and Rhubarb Aglycone plus urokinase group were administrated with Rhubarb Aglycone （lavaged volume was 1 mL per 100 g）, and in sham operation group, model group and urokinase group were lavaged with saline of the same volume. Rats in other groups were given with medicine respectively through artery using catheter at hours 3, 6 and 9 after operation except sham operation group. Rats in urokinase group and Rhubarb Aglycone plus urokinase group were given with urokinase （volume was 20 μL）, and in model group and Rhubarb Aglycone group were administrated with saline of the same volume. （4）At hour 24 after administration through artery, pathological injury of cerebral tissues, brain and stomach hemorrhage ratio, and pathologic changes of brain, lung and stomach were observed, then brain and lung water ratio （BWR, LWR） were measured. RESULTS： 156 rats were dropped out, because of anaesthesia and operation, and 104 rats were involved in the result analysis, （1）brain and stomach hemorrhage ratio： Compared with model group, brain hemorrhage ratio increased in urokinase group at hour 9 （66.67%,28.57%,P 〈 0.05）. Brain and stomach hemorrhage ratio were all higher in 9-h urokinase group than that in 3-h group （brain.66.67% ,18.75% ;stomach.41.18% ,17.65,P 〈 0.05）, and brain hemorrhage ratio was lower in g-h Rhubarb Aglycone plus urokinase group than that in g-h urokinase group （P〈 0.05）. （2）brain, lung and stomach pathologic changes： Compared with sham operation group, pathologic changes of brain, lung and stomach were more obvious in each model group, while the injury of brain and lung in each treatment group lessened compared with corresponding model groups. The injury of brain, stomach and lung in 9-h groups were obvious than that in 3-h groups. In 9-h Rhubarb Aglycone plus urokinase group, the injury became abated compared with urokinase group and Rhubarb Aglycone group, respectively （P 〈 0.05）. （3）BWR and LWR： BWR and LWR in model groups were all higher than that of sham operation group （P〈 0.01 ）. Compared with model group, BWR and LWR in urokinase group and Rhubarb Aglycone plus urokinase group were lower（P〈 0.01 ）. BWR and LWR in each 9-h group increased than that in 3-h group （P〈 0.01 ,P〈 0.05）. BWR in 6-h Rhubarb Aglycone plus urokinase group and LWR in 9-h Rhubarb Aglycone plus urokinase group were all lower than that in corresponding urokinase groups（P 〈 0.05）. CONCLUSION： Delayed thrombolysis therapy can lead to increase of brain and stomach hemorrhage ratio, aggravation of BWR and LWR, obvious pathologic changes of brain, lung and stomach. Rhubarb Aglycone associated thrombolysis therapy can decrease brain hemorrhage ratio, improve changes of neurocyte ultramicrostructure, reduce BWR and LWR, then perform protective effects on lung and stomach caused by cerebral ischemia.