The Experimental Research On Neuroprotective Mechanism Of "Twelve Well-points" Acupuncture Method In Focal Cerebral Ischemic Rats

Objective: To observe the inflammatory reaction and morphologic changes in ischemic brain tissue, related changes of cytokine and adhesion molecule, and the time-effect of the 'Twelve Well-points" acupuncture method on focal cerebral ischemia in ratsMethods: (1) Use photochemical method inducing one-side middle cerebral artery occlusion in rats to make focal cerebral ischemia experimental model. (2) Use Neuroethological index to compare the symptoms of pretreatment and post-treatment, and thereby work out the statistical estimation. (3) Use morphologic method to observe the inflammatory reaction in ischemic brain tissue, especially the phenomena of adhesiveness and infiltration of the inflammatory cells. (4) Use hemorrheologic indexes, erythrocyte deformation and Erythrocyte aggregation to observe the intervention of acupuncture. (5) Use radioimmunoassay method to assay the levels of the BL-1 (3, IL-6, IL-8 and TNF- a in the ischemic brain tissues and me blood serum. (6) Use immunohistochemitty (S-P method) and enzyme-linked immunosorbent assay (ELISA) method to observe the expressions of TGF- P1, ICAM-I, BDNF in ischemic penumbra, which are used to analyze the time-effect of "twelve well-points" acupuncture method.Results: (1) From 3 to 24 hours after acute cerebral ischemia, the inflammatory cell adhered and infiltrated to penumbra of the ischemic cortex, and this phenomenon aggravated as the ischemia continued. After treated by "twelve well-points" acupuncture, the necrosis areas reduced and the degree of the adhesiveness and infiltration of the inflammatory cells also decreased. (2) From 3 to 24 hours after acute cerebral ischemia, the levels of BL-1 P, DL-6, IL-8, TNF- a in the ischemic brain tissues and the levels of the IL-8. TNF-a in blood serum increased significantly, which can be decreased by acupuncture significantly; the level of IL-6 in blood serum increased sharply in 6 hours after ischemia, and retured to normal range from 12 to 24 hours after ischemia Accupuncture can decreased the level of IL-6 during the early ischemic period (from 3 to 12 hours), and increased the level during the late ischemic period (from 12 to 24 hours). (3) From 3 to 24 hours after ischemia, TNF-a expression level in ischemic cortex neurons and ICAM-1 expression level in vascular endothelial cells increased (peaked at 6 hours after ischemia). And at the same time, TGF- P1 and BDNF expression levels in ischemic cortex neurons and glial cells increased, too. Acupuncture can not only decrease the expression of TNF- a and ICAM-1, but also increase the expression of TGF- P 1 and BDNF significantly, and the increase continued as the time pastConclusion: "twelve well-points" acupuncture method can regulate the expression of cytokine and adhesion molecule in ischemic penumbra by influencing the contents of cytokines in brain tissues and blood serum, and thereby reduce the inflammatory reaction and protect function of cranial nerve when cerebral ischemia takes place.To sum up, "twelve well-points" acupuncture method can effectively improve the necrosis degree of the penumbra area in focal cerebral ischemia rats, relieve pathomorphologica] changes of cerebral infarction focus, improve microcirculation and collateral circulation, increase cerebral blood flow, improve energy metabolism, alleviate injury of neurons induced by hypoxia and hypoglycemia, prevent lesions of free radical and NO, decrease intracellular Ca2+ and ICAM-1 expression, and increase TGF- P 1 and BDNF expressions. This research provided experimental foundation to go deep into studying the "twelve well-points" acupuncture method.