【作者】 吴婧；

【导师】 唐涛；

【作者基本信息】 中南大学 ， 中西医结合临床， 2010， 硕士

【摘要】 目的脑出血(intracerebral hemorrhage, ICH)是一种常见的中风亚型,以其高发病率,严重的致死率和致残率而受到人们越来越多的关注。但是目前除了外科手术清除血肿和脱水降颅内压外,尚无其它有效的治疗方法。足三里为足阳明胃经合穴,目前关于针刺足三里在脑出血中的实验和临床研究主要集中于抑制神经细胞凋亡,促进神经功能恢复等方面。本研究通过观察针刺足三里对脑出血后缺氧诱导因子-1α(hypoxia-inducible factor-1α, HIF-1α)和血管内皮生长因子(vascular endothelial growth factor, VEGF)表达的影响,初步探讨针刺足三里治疗脑出血的作用机制。方法120只SD大鼠随机分为假手术组(30只)、假针刺组(30只)、脑出血组(30只)、针刺组(30只),通过立体定位向脑内苍白球注入Ⅶ型胶原酶0.5U建立脑出血模型。假手术组注入等体积生理盐水。针刺组针刺双侧足三里,假针刺组针刺大鼠双侧非经非穴处皮肤。采用免疫组化方法观察各组3、7、14天HIF-1α和VEGF表达的空间位置变化和蛋白表达情况；运用实时荧光定量PCR (Real-time polymerase chain reaction, RT-PCR)检测各组造模术后3、7、14天各时间点HIF-1α和VEGF mRNA表达的动态变化。结果假手术组不同时间点HIF-1a和VEGF未见明显表达。脑出血组、假针刺组和针刺组3天时在血肿周围均可观察到大量HIF-1α和VEGF阳性微血管,7天HIF-1α和VEGF阳性微血管开始深入血肿区,随后出现在血肿区。脑出血3天时HIF-1α蛋白表达即显著增高,随后逐渐降低。VEGF蛋白表达从3天时即明显升高,一直持续到14天。针刺组HIF-1α蛋白、VEGF蛋白表达水平较脑出血组显著升高(P<0.01),而假针刺组与脑出血组比较无统计学意义。RT-PCR结果显示：假手术组、假针刺组、脑出血组和针刺组HIF-1αmRNA表达水平无明显差异；VEGF mRNA在假手术组未见明显表达,脑出血后3天VEGF mRNA表达即明显增强,一直持续到14天,针刺组VEGF mRNA表达水平在各时间点均明显高于脑出血组(P<0.05),而假针刺组与脑出血组比较差异无统计学意义。结论针刺足三里能促进脑出血大鼠脑内HIF-1α和VEGF表达,可能有利于脑出血损伤区的微血管系统重建,从而实现损伤组织的修复。更多还原

【Abstract】 Objective Intracerebral hemorrhage(ICH) is a common and often fatal stroke subtype, to which an increasing number of people are paying close attention because of its high morbility, mortality and mutilation rate. However, currently there is no medical therapy available for those patients except blood pressure reduction and neurosurgical evacuation of the hematoma. The Zusanli (ST36) is the He-sea acupoint of Stomach Meridian of Foot-Yangming(ST). Clinical and experimental studies on ICH by acupunctrue at the Zusanli are focused on aspects of inhibiting the apoptosis of neurons and promoting neurological functional recovery. To investigate the mechanisms of acupunctrue at the Zusanli (ST36), the expressions of hypoxia-inducible factor-la (HIF-1α) and vascular endothelial growth factor (VEGF) were observed in damaged region of rats’brains after intracerebral hemorrhage (ICH).Methods The ICH models were established by injecting collagenase typeⅦ0.5U stereotaxically into right globus pallidus in rats. Animals were randomly divided into four groups:sham-operation (SO), ICH+NA (EA therapy on non-acupoint), ICH and ICH+EA group (EA therapy on Zusanli). The same volume of saline was given to the SO group. The spatial distribution and protein expression of HIF-la and VEGF in each group were observed by immunohistochemistry on day3、7and14 after modeling. In the meantime, the dynamic changes of VEGF and HIF-la mRNA expressions in all groups were assayed by Real-time polymerase chain reaction (RT-PCR).Results No significant expression of HIF-la and VEGF in the Sham-operative group was found during the experiment. In ICH, ICH+NA and ICH+EA groups, many VEGF immunoreactive microvessels of the dilated outline were detected in the perihematomal tissue from 3 days, and the vessels extended into the clot from the surrounding area at about 7 days, then appeared in the clot thereafter. HIF-la protein level significantly increased from 3 days, then began to decline. The expression of VEGF protein in ICH group increased gradually from day 3 to day 14. But ICH+EA group of HIF-la and VEGF protein expression were significantly higher than that in ICH group at the corresponding time points(P<0.01). There was no statistic significance between ICH+NA and ICH groups.HIF-1αmRNA levels were unchanged in SO, ICH+NA, ICH and ICH+EA group. There was no obvious expression of VEGF mRNA in rat brains of sham-operates groups. The VEGF mRNA expression in ICH group increased gradually from day 3 to day 14, ICH+EA group was significantly higher than ICH group at the corresponding time points(P<0.05). There was no statistic significance between ICH+NA and ICH groups.Conclusions The acupuncture zusanli therapy promote VEGF expression in brain of intracerebral hemorrhagic rats, which may accelerate the angiogenesis in the reconstruction of microvascular network in the damaged zone, and facilitate the repair of damaged tissue.更多还原