论文标题:电针对局灶性脑缺血再灌注模型大鼠免疫炎症反应影响的实验研究
The Experimental Research of Electro-Acupuncture on Inflammatory Immunoreactions in Focal Cerebral Ischemia-Reperfusion Rats
论文作者
论文导师 袁红,论文学位 硕士,论文专业 针灸推拿学
论文单位 北京中医药大学,点击次数 89,论文页数 65页File Size1251K
2007-05-01论文网 http://www.lw23.com/lunwen_375181642/
Electro- Acupuncture;; Focal Cerebral Ischemia–Reperfusion (CIR);; Middle Cerebral Artery Occlusion Model (MCAO);; Inflammatory Immunoreactions;; Interleukin-1β(IL-1β);; Intercellular Adhesion Molecule-1 (ICAM-1)
研究目的 脑血管病是临床常见病,其发病率高、预后较差,现已成为中老年人第一致残和前三位致死原因,严重威胁着人类的健康,甚至生命。近年来,对其发病机制的研究已经有了较大的进展,尤其在缺血性脑血管病方面,大量的动物实验和临床观察资料表明,与脑缺血-再灌注有关的急性炎症反应促进了继发性脑损害的发展,缺血和再灌注早期产生的粘附分子和细胞因子构成了缺血性损伤向炎症性损伤转变的基础。因而通过各种途径抑制炎性因子的表达、减缓免疫炎症损伤的进程,进而减轻由于缺血-再灌注所带来到的脑组织后续损伤就成为我们目前研究的一大重点。 研究方法 本实验采用栓线法闭塞大鼠大脑中动脉,建立局灶性脑缺血再灌注模型(MCAO模型),观察脑缺血再灌注6h、24h两个不同时间点缺血侧脑组织形态学改变和炎症反应情况,外周血和脑组织中IL-1β、ICAM-1含量的变化,以及电针“足三里”、“百会”、“水沟”穴对以上各项指标的影响,以研究针刺疗法对于大鼠脑缺血-再灌注损伤不同时期点免疫紊乱的调节作用,并探讨针刺调节作用的相关作用和机制。(1)应用苏木精-伊红(HE)染色法观察缺血侧脑组织神经元缺血坏死情况,炎症免疫反应改变,炎性细胞黏附、浸润情况;(2)采用ELISA法检测外周血中IL-1β、sICAM-1的含量变化;(3)采用ELISA法检测缺血侧脑组织匀浆中IL-1β、ICAM-1的含量变化。 研究结果 (1)局灶性脑缺血再灌注早期(1-24h)缺血皮质出现神经元变性坏死和炎性细胞聚集、粘附及浸润,脑组织坏死范围和程度随着病程进展,炎症反应的继续而扩大加重。电针可降低炎性细胞粘附、浸润,减缓炎症免疫反应,减轻神经元坏死程度。 (2)在脑缺血再灌注超早期(6h)外周循环血及脑组织中IL-1β、ICAM-1表达水平即明显增高,且二者之间存在着单向递进关系,即炎性细胞因子IL-1β的表达能够激活内皮细胞产生ICAM-1,缺血区中性粒细胞的浸润需要内皮细胞经多个炎性介质的介导和炎性细胞的相互作用才能完成。 (3)电针可降低脑缺血不同再灌时间外周血及脑组织中IL-1β、ICAM-1的含量水平,尤其在缺血再灌后6h时这一作用更为显著。 结论 针刺治疗缺血-再灌注损伤大鼠可通过在一定范围内降低脑缺血后免疫炎症反应中炎性介质的表达,来抑制炎性细胞在缺血区的聚集和激活,减少炎性因子的释放,减缓炎症反应的进程;同时针刺通过减少炎性因子的表达抑制细胞毒性物质的释放和缺血后迟发性神经元的损伤,从而改善了局部微循环,起到了很好的脑保护作用。 综上所述,电针对于脑缺血后免疫炎症反应的调节作用是多方向、多环节、多途径的,它不局限于仅仅对单个炎性介质的向上或向下调节,而是使参与炎症反应的多个细胞因子、黏附因子以及其他炎性介质的表达都趋向于正常值,使整个的神经-内分泌-免疫网络在新的水平达到平衡和统一,从而调整机体紊乱的免疫状态,减轻由于炎症反应所带来的损害。针刺调动机体的自身调节功能来减轻脑缺血-再灌注所带来的炎性损伤,是针刺治疗缺血性脑血管疾病的重要途径之一。
Objective: Cerebrovascular disease with high disability rate, lethality and death rate endanger human health and life seriously. Cerebral ischemic apoplexy accounts for over half clinical classifying at the cerebrovascular disease show a tendency to increase progressively. Many research materials show, acute inflammatory reaction with the Focal Cerebral Ischemia-Reperfusion (CIR) is a kind of dangerous factor to induce brain damage. The adhesion molecules and the cytokines with this process are the foundation of turning ischemic injury into inflammation. Therefore, it is one of the most important subjects of experimental and clinical researches to find the effective methods to prevent the expression of inflammatory factors so as to relieve the injury of inflammatory immunoreactions and brain damage. Methods: The experiment takes the model of acute focal cerebral ischemia and reperfusion by occluding middle cerebral artery (MCAO). We observe the morphologic changes and inflammatory reaction of rats in the model of acute focal cerebral ischemia and reperfusion at 6h and 24h after reperfusion, the changes of the contents of cytokines-- IL-1βand ICAM-1 in the circulating blood and brain tissue, and the influence of electro-acupuncture on acu-points‘Zusanli’,‘Baihui’,“Shuigou”for the above said indexes. The purpose of this experiment is to study the regulatory function of acupuncture therapy upon immunologic derangement at different time points in the rats of the model of acute focal cerebral ischemia and reperfusion, and probe into the correlated effect and mechanism of acupuncture in the therapy of acute ischemic stroke. (1) Observe the injury and necrosis of neuron in the damaged brain tissue, the changes of inflammatory immunoreactions, the adhesion and infiltration of inflammatory cell by hematoxylin-eosin staining(HE); (2) Detect the content of IL-1βand sICAM-1 in the serum by applying the technique of Enzyme-Linked Immunosorbnent Assay. (3) Detect the content of IL-1βand ICAM-1 in the ischemic brain tissues by applying the technique of Enzyme-Linked Immunosorbnent Assay. Results: (1) In the early stage of acute focal cerebral ischemia and reperfusion (1-24h), degeneration and necrosis of neuron, the accumulation、adhesion and infiltration of inflammatory cell occurred. The scope and extent of brain tissue necrosis expanded and deteriorated with progressing of acute focal cerebral ischemia and reperfusion and the process of inflammatory reaction. Electro- acupuncture is able to reduce the inflammatory cell adhesions and infiltrations, and alleviate the condition of inflammatory immunoreactions and neuron necrosis. (2) The expression of IL-1βand ICAM-1 in the circulating blood and damaged brain tissue significantly increased after the process of focal cerebral ischemia and reperfusion, especially at the time of 6h after reperfusion. The results of the experiment indicate that the expression of IL-1βis a stimulus to the endotheliocytes which excrete Intercellular Adhesion Molecule-1 (ICAM-1). IL-1βand ICAM-1 both take part in the process of infiltration of neutrophil and many other inflammatory cells and medium also accelerate this proceeding. (3) Electro- acupuncture can help to reduce the content of IL-1βand ICAM-1 in the ischemic brain tissues and serum at different time points after the reperfusion, especially significant in down regulation at 6h after focal cerebral ischemia and reperfusion. Conclusion: Acupuncture treatment may reduce the expression of the inflammatory cells and medium in a certain scope to prevent the accumulation and activation of inflammatory cells as well as the releases of the inflammatory factors. Meanwhile, acupuncture may also cut the content of cell toxic substances and relieve the delayed neuronal damages, thereby ameliorating the microcirculation, and promoting brain blood supply, contributing to the protect of the brain. To sum up, the general regulation function to the focal cerebral ischemia-reperfusion model rats through electro-acupuncture has the characteristic of multi-direction, multi-links and multi-passes. It not only up-regulation or down-regulation the single inflammatory medium, but also make the cytokines、adhesion factors and many other inflammatory substances, which all take part in the process of inflammatory immunoreactions, trend to the normal value. And it makes the Nerve-Endocrine-Immunity net achieve balance and unify in the new-level. Acupuncture is able to effectively relieve the inflammatory injury of cerebral ischemia and reperfusion, which is one of the most important methods of acupuncture to treat the ischemic cerebrovascular diseases in the clinic.
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