星形胶质细胞钙瞬变在不同方式针刺调节急性脑缺血大鼠神经血管单元的作用研究

The benign regulation of acupuncture on microcirculation disorder may be the internal mechanism of acupuncture with extensive role, but the specific target(s) and the molecular mechanism has not been clarified. We have found that different acupuncture or electroacupuncture with different intensity and frequency could specifically regulate abnormal cerebral blood flow on acute cerebral ischemia in rats with various forms. changes in cerebral blood flow is a reflection of the functional activities of the neurovascular unit, and relate closely to the mode of calcium transient release in astrocyte, so that we speculated that the afferent acupuncture signal with different ways, possibly induced different patterns of calcium transients from astrocyte, and then regulate the neurovascular unit. this project intends to observate dynamicly in vivo spatio-temporal correlation of different methods of acupuncture and the cerebral ischemic area microvascular vasomotor, blood cell movement and peritubular astrocyte calcium release mode, and study the effect of acupuncture on protein molecular activity, such as glutamate receptors, glutamate transporter, etc. which related to astrocytes calcium release, by using acute cerebral ischemia in rats as the model of acute microcirculation disturbance, comprehensive application of methods of two-photon intravital microscopic imaging, immune fluorescence tracer, and molecular immunoblotting, etc., with changing modes of intensity, frequency and position of electroacupuncture and acupuncture manipulation. This study will try to explore the specific targets and molecular mechanism of benign adjustment of acupuncture on brain microcirculation, thus to deepen understanding of the principles of acupuncture effect.

针刺对微循环障碍的良性调整可能是其作用广泛的内在机制，但其具体的作用靶点和分子机制尚未阐明。我们前期发现，不同强度、频率的电针或手法对急性脑缺血大鼠的异常脑血流有不同形式的特异性调节。脑血流的变化是神经血管单元功能活动的反映，且与星形胶质细胞钙释放模式密切相关,故推测不同方式针刺信号传入后,可能通过诱导不同模式的星形胶质细胞钙释放而调节神经血管单元。为此，本项目拟以急性脑缺血大鼠为急性微循环障碍模型，综合应用双光子活体显微成像、免疫荧光示踪、分子免疫印迹等技术，以变换电针强度、频率和部位以及手针操作等方式，活体动态观察不同方式针刺与缺血脑区微血管的舒缩、血细胞运动以及管周星形胶质细胞钙释放模式的时空关联性，并研究针刺对星形胶质细胞上与钙释放有关的谷氨酸受体、谷氨酸转运体等相关蛋白分子活性的影响，以探寻针刺良性整调脑微循环障碍的具体靶点并阐释其分子机制，进一步加深对针刺作用原理的理解。

本项目基于针刺对脑微循环障碍的调整作用广泛，意义重大的假设，围绕不同针刺方式对急性脑缺血大鼠微循环障碍为中心进行了系统的研究，从中获得许多有益于机理认识和对临床应用具有指导意义的研究成果和启示，一方面厘清了对电针与传统手针作用规律和特点的认识，另一方面加深了对针刺调节微循环障碍中星形胶质细胞可能起重要作用的认识，对长时程针刺可能引起脑损伤恶化，导致新证型转化的机理进行了初步探究，研究进一步加深了对针刺作用原理的理解。