犀角地黄汤对局灶性脑缺血再灌注大鼠血管新生及神经再生的实验研究

发布时间：2018-07-07 19:29

本文选题：犀角地黄汤 + 脑缺血再灌注　；参考：《南京中医药大学》2017年硕士论文

【摘要】：目的:探讨犀角地黄汤对局灶性脑缺血再灌注大鼠的脑保护作用及其对血管新生、神经再生相关蛋白表达的影响,为犀角地黄汤治疗脑梗死提供实验依据和药理学基础。方法:将健康雄性SPF级SD大鼠随机分为5组:假手术组(Sham group),模型组(Model group),尼莫地平组(Nim group),犀角地黄汤高剂量组(XJDH-H group),犀角地黄汤低剂量组(XJDH-L group)。动物在造模前1周开始每天给药:犀角地黄汤高剂量组和低剂量组分别灌胃给予22.5g/kg·d和11.25g/kg·d的犀角地黄汤,尼莫地平组给予尼莫地平1mg/kg·d,假手术组和模型组给予等体积的生理盐水,末次给药2 h后进行手术造模。采用线栓法阻塞大鼠中动脉(middle cerebral artery occlusion,MCAO),建立局灶性脑缺血再灌注大鼠模型,造模2h后实现再灌注。24h大鼠自然清醒后,进行神经功能评分,HE染色观察组织形态改变,TTC染色检测脑梗死体积,采用Western Blot检测大鼠缺血脑组织中 VEGF、bFGF、CD34、BDNF、GDNF 的表达。结果:1.犀角地黄汤对脑缺血再灌注大鼠神经行为学的影响假手术组大鼠行为学无明显改变,评分为0分;与假手术组比较,模型组大鼠神经功能评分具有显著性差异(p=0.000)。犀角地黄汤高剂量组神经功能评分在第2天、第3天、第4天较模型组显著降低(p0.01)。犀角地黄汤低剂量组神经功能评分在第2天较模型组显著降低(p0.01)。尼莫地平组神经功能评分在第2天、第3天较模型组显著降低(p0.01,p0.05)。2.犀角地黄汤对脑缺血再灌注损伤大鼠脑组织病理学的影响组织形态学结果显示,假手术组的大鼠脑组织细胞形态规则,连接紧密。模型组神经细胞结构发生严重改变,形态结构紊乱,大小不等,细胞核深染固缩,核仁、核膜不清,神经细胞周围间隙增宽。各给药组可见神经细胞缺血性改变均减轻。其中犀角地黄汤高剂量组神经细胞缺血性改变明显减轻。3.犀角地黄汤对大鼠脑缺血再灌注损伤脑梗死体积的影响假手术组的脑组织无梗死。与假手术组相比,模型组大鼠可见明显的脑梗死。与模型组相比,犀角地黄汤高剂量组、尼莫地平组脑梗死体积明显缩小(P0.01,P0.05),犀角地黄汤高剂量组效果优于尼莫地平组(P0.05)。犀角地黄汤低剂量组脑梗死体积缩小不明显(P0.05)。4.犀角地黄汤对脑缺血再灌注大鼠血管新生相关蛋白表达的影响在假手术组中,脑组织VEGF、bFGF、CD34的表达较低。模型组VEGF与bFGF的表达均显著高于假手术组(p0.05),CD34表达无明显差异(p0.05)。与模型组比较,犀角地黄汤低、高剂量组VEGF、bFGF、CD34的表达水平明显升高(p0.05,p0.01),且呈一定的剂量依赖性。尼莫地平组与模型组比较也可显著升高VEGF、bFGF、CD34的表达(p0.01)。5.犀角地黄汤对脑缺血再灌注大鼠神经再生相关蛋白表达的影响与假手术组比较,模型组BDNF、GDNF表达升高不明显(p0.05);与模型组比较,犀角地黄汤高剂量组和尼莫地平组BDNF、GDNF水平明显升高(p0.01)。犀角地黄汤低剂量组BDNF水平明显升高(p0.05),但GDNF升高不明显(p0.05)。结论:犀角地黄汤可改善脑缺血再灌注后大脑的组织形态学变化,缩小脑梗死面积,对脑缺血再灌注损伤具有保护作用,其机制可能与上调VEGF、bFGF、CD34、BDNF、GDNF的表达,促进神经和血管再生相关。
[Abstract]:Objective: To explore the protective effect of rhinoceros Dihuang Decoction on cerebral ischemia reperfusion in rats and its effect on angiogenesis and expression of nerve regeneration related protein, and provide experimental basis and pharmacological basis for the treatment of cerebral infarction by rhinoceros Dihuang Decoction. Methods: 5 healthy male SPF SD rats were randomly divided into 5 groups: sham operation group (Sham group), model Group (Model group), nimodipine group (Nim group), high dose group (XJDH-H group) of rhinoceros Dihuang Decoction (XJDH-H group) and low dose group (XJDH-L group) of rhinoceros Dihuang Decoction (XJDH-L group). The high dose group and low dose group of rhinoceros Dihuang Decoction group and low dose group were given the rhinoceros horn Rehmannia Soup for 22.5g/kg D and 11.25g/kg D, and nimodipine group was given to nimodipine group. The modipine 1mg/kg D, the sham operation group and the model group were given equal volume of physiological saline, and the last dose was 2 h after the operation. The rat middle artery (middle cerebral artery occlusion, MCAO) was blocked by the thread thrombus method. The rat model of focal cerebral ischemia reperfusion was established. After the model 2H was created, the reperfusion of.24h rats was naturally awake and nerve was carried out. HE staining was used to observe the changes of tissue morphology, TTC staining was used to detect the volume of cerebral infarction, and the expression of VEGF, bFGF, CD34, BDNF and GDNF in the ischemic brain tissue of rats was detected by Western Blot. Results: the neurobehavioral effects of 1. rhinoceros Dihuang Decoction on cerebral ischemia reperfusion rats were not significantly changed in the sham operation group, and the score was 0. The nerve function score of the model group was significantly different (p=0.000). The nerve function score in the high dose group of the rhinoceros Dihuang decoction was second days, third days and fourth days compared with the model group (P0.01). The nerve function score of the low dose group of rhinoceros Dihuang decoction was significantly lower than the model group (P0.01) in the second day. Nimodipine group's nerve function The score in second days and third days was significantly lower than that of the model group (P0.01, P0.05). The histopathological results of.2. rhinoceros Dihuang Decoction on cerebral ischemia reperfusion injury in rats showed that the cell morphology of the rat brain tissue in the sham operation group was regular and closely connected. The nuclear deep staining, nucleolus, nucleolus, nuclear membrane unclear, the peripheral space of the nerve cells widened. The ischemic changes of nerve cells in each group were reduced. The ischemic change of the nerve cell in the high dose group of rhinoceros Dihuang Decoction obviously alleviated the effect of.3. rhinoceros Dihuang Decoction on the volume of cerebral ischemia reperfusion injury in rats. Compared with the model group, the cerebral infarction volume in the nimodipine group was significantly reduced (P0.01, P0.05), and the effect of the high dose group of rhinoceros Dihuang decoction was better than that of the nimodipine group (P0.05). The effect of P0.05.4. rhinoceros Dihuang Decoction on the expression of angiogenesis related protein in cerebral ischemia reperfusion rats was lower in the sham operation group, and the expression of VEGF, bFGF and CD34 in the brain tissue was lower. The expression of VEGF and bFGF in the model group was significantly higher than that of the sham operation group (P0.05), and the expression of CD34 was not significantly different (P0.05). The expression level of VEGF, bFGF and CD34 increased significantly (P0.05, P0.01), and was dose-dependent. Nimodipine group and model group could also significantly increase VEGF, bFGF, CD34 expression (P0.01).5. (P0.01).5. rhino Dihuang Decoction on the expression of neural regeneration in cerebral ischemia reperfusion rats compared with the sham operation group, the model group BDNF, GDNF. The elevation of the expression was not obvious (P0.05). Compared with the model group, the level of BDNF and GDNF in the high dose group of rhinoceros Dihuang Decoction and the nimodipine group increased significantly (P0.01). The level of BDNF in the low dose group of rhinoceros Dihuang decoction was significantly increased (P0.05), but the increase of GDNF was not obvious (P0.05). Conclusion: rhinoceros Dihuang Decoction can improve the histomorphological changes of the brain after cerebral ischemia reperfusion. Reducing the area of cerebral infarction has protective effect on cerebral ischemia reperfusion injury. Its mechanism may be related to the up regulation of VEGF, bFGF, CD34, BDNF, GDNF, and the promotion of nerve and vascular regeneration.
【学位授予单位】：南京中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R285.5

【参考文献】