针刺对缺血性脑损伤神经血管单元功能的影响及机制研究

发布时间：2018-01-03 23:08

本文关键词：针刺对缺血性脑损伤神经血管单元功能的影响及机制研究　出处：《山东中医药大学》2013年博士论文　论文类型：学位论文

【摘要】：随着人口老龄化的进展，缺血性脑血管病已成为严重危害人类健康的疾病之一。针刺具有安全、简便、经济等优点，不仅在我国接受针灸治疗的脑卒中病人有很高的比例，在国外接受针灸治疗的中风患者人数亦在逐年增多，针刺疗法已经逐渐成为世界范围内中风的常规治疗方法。研究发现，脑缺血的病理生理变化十分复杂，涉及到星形胶质细胞、微血管内皮细胞及神经元损伤及功能失调，并因此提出“神经血管单元”的概念。为研究针刺治疗中风的机制，有必要从神经血管单元角度对其进行进一步探讨，这将为其机制研究提供一个全新的思路，为其临床的应用提供坚实的实验基础。目的：电针MCAO大鼠内关穴、曲池穴，研究针刺对星形胶质细胞激活的影响，探讨针刺调节脑微循环血流量及能量代谢的作用机制，为针刺治疗缺血性脑中风的神经血管单元机制研究提供一定的理论和实验基础。方法：线栓法建立MCAO大鼠模型，通过神经行为学评分、微循环学、组织形态学、免疫组织化学、实时定量PCR、Western blot等方法，观察连续针刺14天对MCAO大鼠神经功能恢复、软脑膜微循环血流量，缺血脑组织形态改变、脑组织谷氨酸含量、胶质纤维酸性蛋白（GFAP）、代谢性谷氨酸受体5（mGluR5）mRNA、花生四烯酸细胞色素P450表氧化酶2C11（CYP2C11）mRNA表达的影响，探讨针刺通过活化星形胶质细胞调节微血管血流量的作用及机制；通过观察针刺对MCAO大鼠乳酸及丙酮酸含量、脑组织GFAP蛋白的表达、葡萄糖转运体1（GLUT1）、单羧酸转运体MCT1、单羧酸转运体MCT2的表达的影响，从神经血管单元功能角度探讨针刺通过激活星形胶质细胞调节脑组织微循环血流量及能量代谢的作用及机制。应用SPSS19.0统计学软件进行数据统计及结果分析。结果：针刺内关穴、曲池穴能够提高MCAO大鼠神经体征评分，，提高脑组织中GFAP、MCT1、MCT2及GLUT1蛋白的表达；上调GFAP mRNA、mGluRs mRNA、CYP2C11mRNA的表达。针刺内关穴、曲池穴可以有效减少MCAO大鼠脑组织中过量的谷氨酸。结论：针刺可激活星形胶质细胞，一方面可促进mGluRs及CYP2C11mRNA的表达，增加大鼠软脑膜微循环血流量；另一方面，可以促进无氧糖酵解产物乳酸的利用，增强缺血脑组织对葡萄糖的摄取，以满足神经元在脑缺血时的能量需求；针刺对缺血脑组织神经血管单元成员的功能具有调节作用，这是其治疗缺血性脑中风的机制之一。
[Abstract]:With the progress of population aging, ischemic cerebrovascular disease has become one of the serious diseases endangering human health. Acupuncture is safe, simple, economic and other advantages, not only in China to receive acupuncture treatment of cerebral apoplexy patients have a high proportion in the number of people receiving foreign stroke patients with acupuncture in the acupuncture therapy also increased year by year. Has become a routine treatment in the world stroke.
The study found that the pathophysiological changes of cerebral ischemia is very complex, involving astrocytes, microvascular endothelial cells and neuronal injury and dysfunction, and therefore put forward the concept of "neurovascular unit". To study the mechanisms of acupuncture in treatment of stroke, it is necessary to carry out further discussion from the angle of neurovascular unit, which will provide a new idea for the research of its mechanism, to provide experimental foundation for the clinical application.
Objective: MCAO rats electroacupuncture Neiguan, Quchi, effect of acupuncture on the activation of astrocytes and explore the mechanism of acupuncture to regulate cerebral microcirculatory blood flow and energy metabolism, provide a theoretical and experimental foundation for the research unit mechanism of neurovascular acupuncture treatment of ischemic stroke.
Methods: the MCAO rat model was established with suture method, score, neural behavior through the microcirculation, morphology, immunohistochemistry, real-time quantitative PCR, Western blot, observed 14 days of continuous acupuncture on nerve function recovery of MCAO rats, the microcirculatory blood flow of pia mater, pathological changes of cerebral ischemia, glutamate content in brain tissue, glial fibrillary acidic protein (GFAP) and metabotropic glutamate receptor 5 (mGluR5) mRNA, four arachidonic acid cytochrome P450 oxidase 2C11 (CYP2C11) mRNA expression and the effects of acupuncture on the activation of astrocytes by regulating effect and mechanism of vascular blood flow; through the observation of acupuncture on MCAO rats of lactic acid and pyruvic acid content the expression of GFAP in brain tissue, protein, glucose transporter 1 (GLUT1), monocarboxylate transporter MCT1, monocarboxylate transporter MCT2 expression, from the perspective of neurovascular unit function by Acupuncture The role and mechanism of over activation of astrocytes in regulating microcirculation, blood flow and energy metabolism of brain tissue were analyzed by SPSS19.0 statistical software.
Results: Acupuncture Neiguan, Quchi acupoint can improve the neurological score of MCAO rats, increase GFAP and MCT1 in brain tissue, the expression of MCT2 GLUT1 protein and up regulation of GFAP mRNA, mGluRs; mRNA, CYP2C11mRNA. The expression of acupuncture Neiguan, Quchi acupoint can effectively reduce MCAO rat brain tissue in excess of glutamate.
Conclusion: acupuncture can activate astrocytes, on the one hand, can promote the expression of mGluRs and CYP2C11mRNA, increase the pial microcirculatory blood flow in rats; on the other hand, use can promote the anaerobic glycolysis of lactic acid, enhanced uptake of glucose in the brain tissue ischemia, in order to satisfy the neurons in the cerebral ischemia energy demand function; effect of acupuncture on ischemic brain neurovascular unit members has a regulatory role, it is one of the mechanisms of treating ischemic stroke.

【学位授予单位】：山东中医药大学
【学位级别】：博士
【学位授予年份】：2013
【分类号】：R245

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