摘要
背景:脑内肿瘤坏死因子α可通过诱导脑缺血再灌注时内皮细胞黏附分子的表达促进炎症反应及血栓形成。目的:观察头穴百会透曲鬓对缺血性脑损伤的良性调节,以及对脑内肿瘤坏死因子α过度表达的抑制作用。设计:随机对照实验。单位:青岛大学医学院附属医院急诊神经科。材料:实验于2002-08/2003-08在青岛大学医学院脑血管病研究所完成。清洁级雌性SD 大鼠15只,体质量230~270g,随机分为假手术组、针刺组和对照组,5只/组。干预:①针刺组和对照组建立局灶性脑缺血再灌注模型,假手术组手术步骤同对照组,但不阻塞大脑中动脉。针刺组在缺血后10m in 给予针刺治疗,在大鼠头顶部相当于人体百会、风府穴常规剪毛消毒,用0.35m m×40m m 毫针在皮下百会穴处左前下方相当于人体曲鬓穴处平刺,进针深度约0.8cm 。另于风府穴直刺一针,深度约0.5cm 。连接全能脉冲电疗仪,百会穴接阳极,风府穴接阴极,电针频率7HZ,强度6m A,时间30m in。②脑片制备及肿瘤坏死因子α原位杂交检测:各组大鼠于缺血2h 再灌注12h 后深麻醉,左心室插管经升主动脉快速灌注生理盐水200m L,再持续灌注质量浓度40g/L多聚甲醛300m L,断头取脑,相同固定液后固定1h。自视交叉处开始向后取冠状切片,片厚约7μm 。肿瘤坏死因子α阳性细胞胞浆着色呈棕黄色,阴性对照细胞不着色。应用VIDAS-21图像分析系统进行图像分析处理,结果用吸光度表示。③缺血脑组织病理学观察:常规苏木精-伊红染色,光镜下观察针刺组和对照组病变区病理变化。主要观察指标:①各组脑缺血再灌注后脑皮层及纹状体内肿瘤坏死因子α的表达。②针刺组和对照组缺血脑组织病理变化。结果:15只大鼠全部进入结果分析。①脑缺血再灌注大鼠脑皮质及纹状体内肿瘤坏死因子α的表达:与假手术组比较,对照组和针刺组表达明显增强(0.302±0.04,0.320±0.02;0.466±0.08,0.423±0.02;0.367±0.03,0.362±0.02;P <0.05);与对照组比较,针刺组表达明显降低(P <0.05)。②缺血脑组织病理变化:对照组脑组织变性坏死程度较重,神经元变性坏死,胶质细胞呈空泡样变性,血管内有明显的白细胞聚集及附壁现象,血管周围亦有明显白细胞浸润;针刺组仅有少量神经元变性坏死及胶质细胞空泡样变性,血管内白细胞聚集减少。结论:脑缺血再灌注后脑皮层及纹状体内肿瘤坏死因子α水平升高,头曲鬓穴处平刺可明显抑制其表达,从而有效阻止炎性细胞因子过量表达及其生物学作用的发挥,减少脑缺血后的炎性损伤。
BACKGROUND: Tumor necrosis factor-alpha (TNF-α) in the brain can promote inflammative reaction and thrombosis through inducing the expression of endothelioal adhesion molecule during cerebral ischemic reperfusion. OBJECTIVE: To investigate the good regulation of scalp point through point acupucture with Baihui (GV-20) to Qubin (GB-7) to ischemic cerebral injury, and to inhibition of TNF-α over-expression within the brain as well. DESIGN: A randomized controlled trial. SETTING: Emergency Neurological Department of the Hospital Affiliated to Medical College of Qingdao University. MATERIALS: The experiment was completed from August 2002 to August 2003 at the Institute of Cerebrovascular Disease in Medical College of Qingdao University. A total of 15 healthy female SD rats of clean grade, weighing 230-270 g, provided by Shanghai Experimental Animal Center of Chinese Academy of Sciences, were at random divided as sham operation group, acupuncture group and control group, with 5 rats in each group. INTERVENTIONS: ① The focal ischemia/reperfusion models were set up for rats in acupuncture group and control groups, the procedur for rats in sham operation group was similar to those in control group, but without obstruction of the middle cerebra artery. After ten minutes ischemia, the rats in acupuncture group were given acupuncture treatment. The hairs in parts correspondent to Baihui (GV-20) and Fengfu (GV-16) points in human were cut down and conventionally sterilized, then a filiform needle in 0.35 mm×40 mm was used to point to point acupucture correspondent to Baihui and Qubin in human to the depth of 0.8 cm. Another needle was used to perpendicularly insert at Fengfu (GV-16) point to the depth of 0.5 cm. A pulse electric machine was applied for 30 minutes, with the positive electrode connecting to Baihui (GV-20) and negative electrode to the Fengfu (GV-16), the frequency being 7 Hz, The electric current being 6 mA. ② Preparation of cerebral section and in situ hybridization assay of TNF-α: All rats of every group, after treatment of 2 hours ischemia and 12 hours reperfusion, were deeply anesthetized, quickly given 200 mL normal saline through the catheterized ascending aorta, and continually given 300 mL paraformaldehyde of 40 g/L concentration; then they were decapitated for collecting the brain, the brain was fixed for 1 hour with the same solidification solution. The coronal section about 7 μm was prepared starting from the part of optic chiasm towards the back. The cytoplasm of TNF-α positive cell was stained brownish yellow, and that of TNF-α negative control cell was not stained. VIDAS-21 image analysing system was used to analyse the image, the result was expressed as absorbance. ③ Pathological observation of ischemic cerebral tissue: After conventional staining with hematoxylin-eosin, the pathological changes of focal areas in both acupuncture group and control group were observed under light microscope. MAIN OUTCOME MEASURES: ① The expression of TNF-α in the cortex and striate body of brain in rats of each group. ② The pathological changes of ischemic cerebral tissues in acupuncture group and control group. RESULTS: All 15 rats entered the final result analysis. ① Expression of TNF-α in the cortex and striate body of brain in rats with ischemic reperfusion: Compared with sham operation group, the expressions in acupuncture group and control group were significantly increased (0.302±0.04, 0.320±0.02; 0.466±0.08, 0.423±0.02; 0.367±0.03, 0.362±0.02; P 〈 0.05); compared with acupuncture group, the expression in control group was significantly decreased (P 〈 0.05). ② Pathological changes of ischemic cerebral tissues: In control group the extent of degeneration and necrosis was more serious, the neuron was degenerative necrosis, the neuroglia cell was in vacuolar degeneration, there was obvious phenomenon of leukocytic aggregation and margination within the vessel, and also leukocytic infiltration around the vessels. In acupuncture group there were only a small nub of neurons being in degenerative degeneration and neuroglia cells in vacuolar degeneration, and less leukocytic aggregation within the vessel. CONCLUSION: The level of TNF-α in the cortex and striate body of brain was increased after ischemic reperfusion. Scalp point through point acupuncture with Baihui (GV-20) to Qubin (GB-7) could markedly inhibit its expression, so that the therapy could effectively check over-expression of inflammatory mediators and their biological effects, hence reducing the inflammatory injury after cerebral ischemia.
出处
《中国临床康复》
CSCD
北大核心
2005年第25期218-220,共3页
Chinese Journal of Clinical Rehabilitation