从稳定细胞外基质探讨健脾补土法抗脑缺血损伤机制

Extra cellular matrix（ECM）is the living environment of neuron, it provide supporting, nutrition, protecting for neuron, it have some similarity as the carry and breeding of spleen in traditional Chinese medicine. Protecting ECM by the method of Invigorating the Spleen opens new avenues for protecting neurocyte, reducing damages of cerebral ischemia.Using cerebral ischemia-reperfusion rats models and hypoxic injured olfactory ensheathing cells models, taking ECM as a key, this project will base on our previous working basis that the method of Invigorating the Spleen can improve the clinical symptoms, reduce Neuron loss, increase the express of LN and decrease the express of MMPs-9 to systematically study the protective effects on cerebral histopathology, blood brain barrier, ECM in Brain Tissue and on the expression of matrix metalloproteinase and it's inhibitor(tissue inhibitor of meta11oproteinase-1,TIMPs), plasminogen activator(PA)and it's inhibitor(PAI) of Invigorating the Spleen Therapy composition. Among the cellular signal pathways, NF-κB pathway is the decisive pathway for ECM degradation. So we will investigate the relationship between the effects on protecting ECM and regulateing NF-κB pathway of Invigorating the Spleen Therapy furthermore.Contrasting the similar betweem ECM and spleen in traditional Chinese medicine, using orientation analogy law, we put forward the invigorating the Spleen therapy, emphasizing the therapy of invigorating the Spleen-the acquired, which help us provide new rules of treatment and therapy for cerebral ischemia disease. Emphasizing the external environment of neuron- ECM , which provide new idea and target for preventing and curing cerebral ischemia disease. The study will have the potentials to develop new Chinese medicine method, and improve the efficacies of Chinese medicine in post-stroke treatment.

神经细胞外基质（ECM）对神经细胞有支撑、营养与保护作用，与中医脾土"主承载"，滋生万物的功能相似，健脾补土法保护ECM为抗脑损伤开辟了新方向。本项目在前期健脾补土法改善临床症状，减少神经元丢失，增加层粘连蛋白表达和减少基质金属蛋白酶（MMPs）-9的表达的基础上，采用脑缺血大鼠模型和嗅鞘细胞缺氧损伤模型，以ECM为切入点研究健脾补土法组方对脑组织病理形态、血脑屏障、脑组织ECM的保护作用以及对MMPs及其抑制剂(TIMPs)、纤溶酶原激活物(PA)及其抑制剂(PAI)表达的影响，进一步从影响ECM降解的NF-κB通路探讨健脾补土法保护ECM的机理。本研究将ECM与中医脾土的若干相似点进行对照，运用取类比象思维，提出健脾补土法治法，重视培补后天脾土，为脑缺血性疾病提供了新的治法；重视保护ECM，为脑卒中治疗提供了新的思路和靶点，对进一步开发新的中医药治疗方法，提高疗效具有潜在的应用价值。

（1）背景：脑卒中具有发病率高、病死率高、致残率高的特点，已成为我国国民第1位死亡原因，研究脑卒中发病机理及抗脑卒中的中医治则治法和新药已成为研究的热点。目前研究侧重于神经细胞本身而忽视神经细胞外基质（ECM），治法重先天之本而薄后天之本。ECM与中医脾土有诸多相似之处，前期研究表明健脾补土法对神经ECM有保护作用，其机理有待进一步明确。.（2）内容：从整体水平和细胞水平探讨健脾补土法组方脑保护作用以及对ECM、ECM降解相关酶类、NF-κB信号通路的影响，揭示健脾补土法组方保护ECM的机理。.（3）结果：对脑组织病理形态和血脑屏障超微结构具有保护作用，降低血脑屏障通透性（中剂量组11.51±3.59 vs模型组14.94±3.13，整体实验，7d）。升高I/R大鼠脑组织和嗅鞘细胞缺氧模型Col IV（中剂量组89.16±33.01 vs模型组51.95±13.74，整体实验，7d）、LN、FN含量，降低MMP-2（中剂量组71.33±12.84 vs模型组107.06±31.75，整体实验，7d）、MMP-9、tPA、uPA表达，促进TIMP1（中剂量组58.97±8.90 vs模型组35.10±14.04，整体实验，7d）、TIMP2、PAI表达；降低IκK-β、pI-κB-α、P50（健脾补土组0.547±0.192 vs模型组0.948±0.147，细胞实验，48h）、p65（健脾补土组0.590±0.086 vs模型组0.824±0.129，细胞实验，48h）表达，促进IκBα表达，抑制NF-κB/p65 DNA结合活性（健脾补土组0.032±0.007 vs模型组0.051±0.009，细胞实验，48h）。.（4）科学意义：以取类比象的思维将神经ECM与中医脾土进行类比，探讨健脾补土法保护神经ECM抗脑缺血损伤的保护作用，为临床提供新的治则治法——健脾补土法。通过健脾补土法对脑I/R大鼠神经ECM、调节ECM降解的酶系以及调节酶系活化的信号通路的影响的研究，可进一步明确该法防治脑I/R损伤的作用机理，为临床提供新的思路和切入点——保护神经细胞外基质。