脉络宁及葛根素注射液联用对2型糖尿病小鼠脑损伤相关因子表达的影响

【作者】 杨祝仁；

【导师】 李兴广；

【作者基本信息】 北京中医药大学 ， 临床中药学， 2008， 硕士

【摘要】 糖尿病是严重危害人类健康的常见病,已成为仅次于肿瘤和心血管疾病之后的第三大疾病。糖尿病由于体内存在胰岛素抵抗和(或)胰岛素分泌不足,使糖和脂代谢发生紊乱,极易发生大血管和微血管病变,并给大脑带来一系列损害,应引起高度重视。常见2型糖尿病对大脑的损害主要表现为脑血管疾病(主要包括血栓性脑梗死、脑栓塞、短暂性脑缺血发作、腔隙性脑梗死以及出血性脑卒中)和糖尿病性脑病(由于糖尿病糖代谢紊乱导致脑血管改变,损害了中枢神经系统,使大脑在结构、神经生理及神经精神等方面发生病理改变,以获得性认知行为缺陷为特征的糖尿病慢性并发症)。中医药对2型糖尿病脑损伤的防治作用,已被临床认可并广泛使用。深入探讨中医药对2型糖尿病脑并发症的作用机理对于指导临床有着重要意义。脉络宁注射液和葛根注射液在临床常用于2型糖尿病并发症的治疗,对于改善高血糖、高血脂,预防控制微血管和大血管并发症等均有较好疗效,本实验通过观察两药合用的干预作用,旨在探讨脉络宁注射液和葛根素注射液联用防治糖尿病脑损伤的作用机制。1.文献研究:就中医对糖尿病脑损伤的认识;中药治疗糖尿病脑损伤的研究进展;现代医学对于糖尿病脑损伤的发病机制、相关研究方法、以及KKAy小鼠动物模型建立等方面进行了总结和综述。2.实验研究:目的:研究脉络宁注射液与葛根素注射液联用治疗糖尿病小鼠脑损伤可能的作用机制。探讨从中医“毒损脑络”理论出发,利用益气养阴、通络化瘀中药治疗糖尿病脑损伤的作用机制。方法:①将2型糖尿病模型KKAy小鼠分为模型组和治疗组。同龄C57BL/J小鼠作为正常组。治疗组小鼠14周龄开始以腹腔注射的方式给药。隔周测定各组小鼠的体重和血糖,并分别于20、24和28周龄处死小鼠取血,检测生化指标,同时取小鼠脑组织观察其病理形态变化。②采用免疫组化化学法,检测小鼠脑组织中ICAM-1和Caspase-3的表达。③采用酶联免疫吸附法(ELISA)方法,检测小鼠血清中sICAM-1和sE-selectin含量。结果:①KKAy小鼠14周龄以后出现了明显的肥胖、倦怠、多尿、高血糖;20周以后出现高脂血症;28周出现脑组织形态学改变;脉络宁注射液与葛根素注射液联用治疗后,治疗组体重增长减缓,血糖降低,血脂受到控制,KKAy小鼠脑组织病理改变减轻。②KKAy小鼠脑组织中ICAM-1和Caspase-3表达增多。脉络宁注射液与葛根素注射液联用治疗后,表达有所降低。③KKAy小鼠血清中sICAM-1含量显著增加, sE-selectin含量无改变。结论:①脉络宁注射液与葛根素注射液联用可以改善KKAy小鼠的体重、高血糖、高血脂,减轻脑组织的病理损伤。②脉络宁注射液与葛根素注射液联用可以减少粘附分子ICAM-1的表达及血清中含量。同时可以减少Caspase-3的表达。这种对血管内皮细胞的粘附分子ICAM-1及凋亡级联反应中关键因子Caspase-3的控制作用,可能是两种中药抗糖尿病脑损伤的作用机理。更多还原

【Abstract】 Diabetes mellitus (DM) is a common disease that seriously hurts people’s health. And it is regarded as the third leading cause of death following tumor and cardiovascular disease.Diabetic mellitus makes the Lipid and glucose metabolic disorder , that is highly susceptible to macrovascular and microvascular diseases. Diabetic mellitus is caused by Insulin Resistance and absolutely or relatively absence of insulin so that affects the brain. Above all, it is necessary to pay attention to the harm of Diabetic mellitus.The injury of the brain which is affected by Type 2 diabetes mellitus is cerebrovascular disease and diabetic encephalopathy.Traditional Chinese Medicine control for Brain Injury of type 2 diabetes mellitus is approved by clinics and widely used.To further study the curative effect of Chinese medicine in treating complication of type 2 diabetes mellitus has great importance to the clinical guidance.Mailuoning injection combines Puerarin injection was used to treat complication of type 2 diabetes mellitus in clinic , they can reduce hyperglycemia, hyperlipemia and control the macrovascular and microvascular diseases. We combine the Mailuoning injection combines Puerarin injection to investigate the mechanism of control for Brain Injury of diabetes mellitus.1.Literature research:According to the Chinese medicine to diabete’s understanding,the Chinese medicine treats diabetes’research development:The modern medicine the pathogenesis which damages to diabetes and related technique, KKAy mice animal model’s aspects and so on establishment,carry on the summary and the review.2.Experiment research:Objective:Investigate the mechanism of Mailuoning injection combines Puerarin injection to treat Brain Injury of diabetes mellitus. Discuss the specific mechanism of Chinese medicine which supplementing qi and nourishing yin、removing blood stasis and activating collaterals in treatment of Brain Injury of diabetes mellitus.Methods:①KKAy mice were randomly divided into two groups: model and treatment group. C57BL/J mice were used to be controls. Treatment group were given by peritoneal from fourteen weeks old. Weight and fasting plasma-glucose were measured fortnightly. They were sacrificed at 20, 24 and 28weeks of age respectively. Serum creatinine, urea, lipids were measured, the changes of the pathomorphology was also be observed.②ICAM-1 and Caspase-3 were detected in cerebral of mice tissue by immunohisto-chemical method.③sICAM-1 and E-selectin were detected in blood of mice tissue by ELISA method. Results:①KKAy mice developed obesity, hyperglycemia after 14 weeks age; developed hyperlipemia at 20 weeks of age; developed the changes of the pathomorphology at 28 weeks of age. After the treatment of Mailuoning injection combines Puerarin injection, the weight of KKAy mice was declined, the hyperglycemia was declined and the hyperlipemia was controlled, the changes of the pathomorphology was also declined.②The expression of ICAM-1 and Caspase-3 in brain of KKAy mice were upregulated. while after the treatment of the Mailuoning injection combines Puerarin injection, the expression was inhibited.③The content of sICAM-1 in blood of KKAy mice was increased, and the content of sE-selectin was invariable.Conclusion:①Mailuoning injection combines Puerarin injection improved weight, hyperglycemia, hyperlipemia and reduce the pathology damage of brain.②Mailuoning injection combines Puerarin injection inhibited the expression of ICAM-1 and Caspase-3. The control action of ICAM-1 -the vascular endothelial cells adhesion molecule and apoptosis cascades reaction’s key effector Caspase-3 maybe the mechanism of control for Brain Injury of diabetes mellitus.更多还原