【作者】 李京京；

【导师】 东贵荣；

【作者基本信息】 黑龙江中医药大学 ， 针灸推拿学， 2008， 博士

【摘要】 目的:通过头穴透刺对脑出血大鼠模型治疗的实验研究,探讨针刺对脑水肿调控因素影响的机制。方法:本实验用健康雄性(wistar)大鼠144只,体重250-300克,随机数字表分为A、B、C三组,每组又各分为12h、24h、3d、7d四个小组。每小组每时间点各12只动物。动物经10%水合氯醛腹腔麻醉(400mg/kg体重),头顶备皮后将动物固定于三维立体定位仪上,局部皮肤处理后以前囱为坐标原点,向大鼠尾侧0.2mm,大鼠右侧2.9mm,确定注射点坐标,用牙科钻钻孔(直径1.0mm)(右尾状核位置),A组:对照组(48只)注入2u10.9%的生理盐水,B组和C组:模型组(48只)和针刺组(48只)均用微量注射器吸取含0.5u/ul胶原酶的生理盐水和7u/ul肝素的生理盐水各1.0ul注入,每组于注射完毕、退针后,缝合头皮,并于12小时、24小时、3d、7d参照Longa FZ五级评分法进行神经功能学评分,评分完毕后,按要求处置,测定脑含水量,进行大鼠病理形态学观察,组织形态超微结构观察及免疫组化方法观察AQP-4,MMP-2和MMP-9的表达。结果:1、脑出血后大鼠出现神经功能缺损,72h达高峰,之后有所下降。针刺组与模型组比较差异显著(P＜0.05)。针刺可以降低神经功能缺损程度。2、脑出血大鼠模型24h后脑含水量明显增高,3d达高峰,针刺治疗有较明显的抗水肿作用,各时间点比较有统计学意义(P＜0.05)。3、脑出血周边组织EB含量在24h达到高峰,24h-72h间维持较高水平,显著高于对照组;针刺组EB含量各时点均显著低于模型组(P＜0.05),EB含量与脑组织含水量呈正相关。4、HE染色显示:脑出血后24h血肿周围可见白细胞浸润,细胞间隙和血管周围间隙增宽,3d上述病变加重,并见大量神经细胞变性坏死,7d仍有增生的胶质细胞,从病变的程度和范围看,针刺组的病理改变要轻于模型组。5、电镜观察BBB超微结构,出血后各时点BBB结构均有破坏,72h最为严重;针刺治疗后BBB的开放相应减小。6、对照组有极少量AQP-4阳性细胞表达;模型组中3d达高峰,各时点均显著高于对照组;针刺组表达减少(P＜0.01)。AQP-4表达与脑组织EB含量呈正相关。7、MMP-2在对照组极少量表达;模型组中24h达高峰,各时点均显著高于对照组;针刺组表达减少(P＜0.01)。8、对照组有少量MMP-9阳性细胞表达;模型组3d达高峰,各时点均显著高于对照组;针刺组表达减少(p＜0.01)。结论:1、大鼠脑出血后AQP-4与MMP-2,MMP-9的释放与激活,是脑出血后早中期脑水肿形成的重要因素之一,导致BBB通透性增加而造成脑水含量增加。2、针刺治疗具有抗脑出血后脑水肿的作用,机制之一是对AQP-4,MMP-2,MMP-9表达的抑制,使BBB开放程度减小,降低脑组织含水量,减轻脑水肿程度。3、丰富头穴透刺治疗急性高血压性脑出血的理论依据。更多还原

【Abstract】 Objective: To research the influence of control factor in rats with brain edema after acute intracerebral hemorrhage treated by scalp penetration acupuncture.Method: healthy male wistar rats (weight:250-300g) were randomly divided into three groups: A,B and C, each group were randomly divided into 12h、24h、3d、7d four time points, the rats were prepared by rules. The rats in group A were injected 0.9% physiological saline 2ul, the rats in group B and C were injected 0.9% physiological saline with 0.5u/ul collagenase 1ul and 0.9% physiological saline with 7u/ul heparin 1ul, then observe the information and data at assignment time : Neurological deficit scale ,the brain tissue water content, brain tissue patho-form, brain tissue ultramicro- texture and AQP-4,MMP-2,MMP-9 by immunity histochemistry method.Results: 1 .the recordings indicate there is Neurological deficit scale in hemorrhage rats, the peak occurred at 72h and descent then. There are significant disparity between model group and control group.2. The brain tissue water content step up after 24h and the peak occurred at 3d, there are significant disparity between model group and control group.3. The content of EB in brain tissue reached the peak at 24h and keep high level during 24h-72h, there are significant disparity between model group and control group. There are positive correlation between brain tissue water content and the content of EB in brain tissue.4. Light-microscope shows there’s severe pathological change in brain hemorrhagic model group, Nerve cells and colloidal cells swell, and in cell space there’s vacuoles of different sizes and the cell space increases. In treating group, hemorrhage and cell dropsy are distinctly Alleviated.5. Electro-microscope shows compared with normal rats, brain hemorrhagic group shows many changes. Especially in the BBB, mitochondria ranges vague and disappear, mitochondria swell, karyoplasm dilutes compared with normal one , other cell apparatuses are still swelling. But the state in treated group is better.6.The AQP-4 expressed a lot in model group and reach peak at 3d, there are significant disparity between model group and control group (P<0.01) . There are positive correlation between content of AQP-4 and the content of EB in brain tissue.7.There shows MMP-2 expression in model group and reach peak at 24h, there are significant disparity between model group and control group (P<0.01)8. There shows MMP-9 expression in model group and reach peak at 3d, there are significant disparity between model group and control group (P<0.01)Conclusion:1 .The above results shows that the content and changes of AQP-4, MMP-2 and MMP-9 have big relationship with cerebral edema after hemorrhage, they may lead to permeability in BBB and brain tissue water increased.2.The symptom became better after acupuncture , one of the mechanism of anti-edema by acupuncture may be adjust the expression of AQP-4,MMP-2 and MMP-9, diminished break in BBB, degrade brain water content, improved symptom accompanied with cerebral edema.更多还原