【作者】 危玲；

【导师】 周小青；

【作者基本信息】 湖南中医药大学 ， 中医诊断学， 2011， 博士

【摘要】 目的探讨丹龙醒脑方对血管性痴呆(VD)老年肾虚证候的治疗机制,并从不同方面阐明丹龙醒脑方对VD大鼠模型的治疗具有类似雌激素治疗特征。方法从第一周开始对100只SD雌性大鼠进行皮下注射D-半乳糖造成老年模型；第四周加用甲状腺素片混悬液灌胃造成肾阴虚模型；造模到第六周末,把大鼠随机分成两部分,行手术造模。一部分大鼠行假手术,仅分离双侧颈总动脉(CCA)；其余大鼠均采用CCA反复结扎再灌注加剪尾放血方法建立VD大鼠模型。夹闭双侧CCA,反复3次,每次10mmin,中间间隔10mmin,同时距尾尖1cm剪尾放血2ml,术后正常喂养。从术后第3d开始治疗。假手术组大鼠正常饮食饮水,不做其他处理；其余手术大鼠随机分成三组：模型+生理盐水组、模型+丹龙醒脑方组、模型+雌激素组。模型+生理盐水组每天以0.9%生理盐水灌胃；模型+雌激素组每三天腹腔注射一次苯甲酸雌二醇注射液；模型+丹龙醒脑方组每天用中药丹龙醒脑方水煎剂灌胃,连续治疗8W。治疗进行到第7W时,进行水迷宫测试各组大鼠学习记忆功能；治疗8W完成后,处死大鼠,取血进行雌二醇(E2),睾酮(T),前列环素(PGI2),内皮型一氧化氮合成酶(eNOS)检测；取海马区脑组织作石蜡切片、HE染色,进行组织病理形态学检查,计数海马区正常神经细胞数,用Tunel法计海马区凋亡神经细胞数；取海马区脑组织,测Bax,Bc1-2蛋白表达；用RT-PCR技术测ERmRNA结果丹龙醒脑方组,雌二醇组与模型组相比：①水迷宫实验的错误次数、游泳时间减少；②海马区正常神经细胞计数明显增多,凋亡细胞减少；③E2水平提高,T水平无明显变化,E2/T比值增大；④eNOS,PGI2含量增加；⑤Bax蛋白表达减弱,Bcl-2蛋白表达增强；⑥海马ERmRNA表达增强。丹龙醒脑方组疗效与雌二醇组比较,略优于雌二醇组。结论以补肾活血通络法组方的丹龙醒脑方通过与ER结合,发挥类雌激素样作用,舒张血管,增加脑血流量,直接保护海马区神经细胞,从而改善VD大鼠模型痴呆症状。这可能是丹龙醒脑方治疗老年肾虚VD的作用机制之一。更多还原

【Abstract】 OBJECETIVE DanLongXingNao Fang to Vascular dementia (VD) treatment of senile kidney deficiency syndrome mechanism and clarify the different aspects of DanLongXingNao Fang treatment of VD rats treated with estrogen-like characteristics.METHOD The first week to 100 SD female big mouse to carry on the hypodermic injection D-galactose to create the old age model, the fourth week adds with the thyroxin piece suspension fills the stomach to create the kidney Yin deficient model, the seventh week divides into chastically the big mouse 4 groups:Sham-operation group, model+ physiological saline group, model+ DanLongXingNao Fang group, model+ estrogen group. In addition to separation of the sham group, only bilateral common carotid artery (CCA), the other rats in each group were repeated with CCA occlusion and reperfusion plus tails bleeding Methods VD rats. Clipping bilateral CCA, repeated three times, each time 10min, an interval of 10 min, while blood from the tip of the tail end cut 1cm 2ml, after normal feeding. After the technique 3d starts to treat. The sham-operation group normal diet potable water, does not make other processing; The model+ physiological saline group fills the stomach every day by 0.9% physiological saline; Model+ estrogen group inject once every three days of estradiol benzoate injection; Model+ DanLongXingnaoFang group uses traditional Chinese medicine DanLongXingnao Fang water decoctum to fill the stomach every day, treats 8W continuously. Treats 7W to carry on the water labyrinth to test each group of big mouse study memory function; After treating 8W, takes the big mouse hepatorenal, line of hepatorenal hydrometry; Draws blood carries on the estradiol (E2),the testosterone (T), the prostacyclin (PGI2), the bast nitrogen monoxide synzyme (eNOS) examines; Hippocampal brain tissue obtained as paraffin, HE staining, the histopathological examination, count the number of nerve cells in hippocampus of normal, the counting hippocampal normal nerve cell number, counts with the Tunel law perishes weakly the cell number; Takes the hippocampal brain organization, measured that the Bax, Bcl-2 protein expresses, measures the ERmRNA content with the RT-PCR technology.Results DanLongXingNao Fang group, estradiol group compared with the saline group:①the number of errors the water maze test, reduction of swimming time;②hippocampus of normal nerve cell counts increased significantly decreased apoptotic cells;③E2 levels by improving, No significant changes in the level of T, E2/T ratio increases;④eNOS, PGI2 content increased;⑤Bax protein expression was decreased, Bcl-2 protein expression increased;⑥ERmRNA expression of the ippocampus. DanLongXingNao Fang curative effect of group compared with the estradiol group, slightly better than the estradiol group. COUCLUSION The kidney and activating blood and collaterals DanLongXingNao Fang combined with the ER，like estrogen play a role in relaxing blood vessels and increase cerebral blood flow，direct protection of neuronal cells, thereby improving the rat model of dementia VD symptoms. This may be DanLongXingnao Fang VD Treatment of senile kidney deficiency one of the mechanisms.更多还原