【作者】 王海涛；

【导师】 曹晓岚；

【作者基本信息】 山东中医药大学 ， 中西医结合临床， 2012， 博士

【摘要】 目的：探讨糖尿病并脑梗死的病机治法，提出“气虚血瘀-瘀热生毒-毒损脑络”为糖尿病并脑梗死的主要病机，清热解毒益气活血法为其有效治疗方法，并应用临床确有疗效的导师经验方益消复瘫汤为治疗药物。研究糖尿病合并脑梗死的可能机制，观察糖尿病并脑梗死后神经损伤的影响，探讨益消复瘫汤治疗糖尿病并脑梗死的作用及机制。方法：应用SD大鼠高脂高糖饲料喂养4周联合小剂量STZ（30mg/kg）腹腔注射方法建立糖尿病模型，1周后筛选血糖合格（血糖≥16.7mmol/L）大鼠进行线栓法建立局灶性大脑中动脉阻塞（middle cerebral artery occlusion,MCAO）脑梗死模型。应用益消复瘫汤高、低剂量为治疗组，并设假手术对照、单纯脑梗死组、单纯糖尿病组为对照，观察比较各组大鼠死亡率并分析死亡原因，行为学观察进行神经功能评分，脑组织TTC染色法计算梗死容积，干湿法计算脑组织含水量，应用尼氏染色观察海马CA1区神经元损伤，比色法计算脑组织匀浆髓过氧化物酶（myeloperoxidase，MPO）活性观察炎性细胞浸润情况，western blot方法检测脑组织炎症信号通路TLR4及其下游肿瘤坏死因子α（tumor necrosis factorα，TNF-α）、白细胞介素-1β（interleukin-1β，IL-1β）蛋白表达。结果：糖尿病脑梗死组大鼠表现了更重的神经损伤，神经行为学评分、脑梗死容积、脑水肿程度、脑组织炎性浸润程度，TLR4、TNF-α、IL-1β表达均重于各对照组，益消复瘫汤能抑制糖尿病脑梗死大鼠TLR4信号通路，减少TLR4蛋白表达，降低TNF-α、IL-1β等炎症因子表达，降低脑组织MPO活性，减少梗死面积，发挥神经保护作用。结论：利用高脂高糖膳食联合小剂量STZ注射基础上进行线栓可成功建立糖尿病脑梗死模型，益消复瘫汤能抑制糖尿病脑梗死后的TLR4炎性信号通路，减少TLR4、TNF-α、IL-1β表达，降低脑组织MPO活性，减轻神经损伤，发挥脑保护作用。更多还原

【Abstract】 Objective： Study the pathology mechanism and treatment method of cerebralinfarction with diabetic,we put forward the main pathology mechanism "qi deficiency andblood stasis,stasis heat reducing toxicant,toxicant damaging brain collaterals". So thetreatment methods of clearing away heat and toxic materials and supplementing qi andactivating blood circulation maybe a valid therapy,and Yi-Xiao-Fu-tan decoction(YXFTT)maybe valid treatment drug. We investigated the changes and the molecular mechanisms ofcerebral vascular damage and tested the therapeutic effects of YXFTT in diabetic rats afterstroke.Methods: Male Sprague-Dawley rats were fed with high fat and high sugar diet for4weeks,then injected intraperitoneally with low dose of streptozotocin (STZ)(30mg/kg)serves as an alternative animal model for type2diabetes,the diabetes rats (blood glucose≥16.7mmol/L)then underwent permanent middle cerebral artery occlusiont.we treated thestroke rats in diabetic with YXFTT for a week, at the same time,we set sham group,MCAOwithout diabetic group,diabete group with sham operation group to control. Neurologicfunction,cerebral infarct volume, brain water content,myeloperoxidase (MPO) activity andhippocampal CA1neurons injury were measured24h after MCAO. Expression ofTLR4,and the downstream effector molecules tumor necrosis factor-alpha (TNF-α) andinterleukin-1beta (IL-1β) were determined by western blot.Results: The stroke rats with diabetes showed significantly higher mortality, biggerinfarcts, increased cerebral edema, worsened neurological status compared to rats withoutdiabtes. The diabetes rats also showed significantly higher post-ischemic inflammatorymarkers MPO activity, exacerbated proinflammatory gene expression compared to rats without diabetes. In addition, the post-ischemic neuroprotective YXFTT reduced cerebralinfarct area and infarct volume. YXFTT reduced the expression of TLR4and TNF-α andIL-1β. Our results suggest that YXFTT inhibits the TLR4signaling pathway in cerebralischemia with diabetes, which may be a mechanism underlying the YXFTT’sneuroprotection.Conclusions: With high fat and high sugar diet combined with small dose injection ofSTZ,then underwent permanent middle cerebral artery occlusion can be successfullyestablished based on diabetic cerebral infarction model, YXFTT in experiments withdiabetic cerebral infarction can reduce nerve damage, reduce the expression of TLR4signaling pathway, play a protective role.更多还原