益气活血方治疗大鼠脊髓型颈椎病的实验研究

发布时间：2018-01-12 11:14

本文关键词：益气活血方治疗大鼠脊髓型颈椎病的实验研究　出处：《南京中医药大学》2016年硕士论文　论文类型：学位论文

【摘要】：目的：采用急性毒性试验评价临床经验方“益气活血方"的安全性,采用耳廓肿试验、醋酸扭体法研究该方的抗炎镇痛作用,采用大鼠脊髓型颈椎病模型研究该方对大鼠受损脊髓神经功能恢复的影响,阐明其促进脊髓功能恢复的相关作用机制,为中药“益气活血方”的临床应用提供理论基础及指导。方法： 1、20只ICR小鼠随机选取5只为对照组,余15只为实验组,采用最大给药量的方法研究益气活血方的急性毒性反应；2、以二甲苯致小鼠耳肿胀法以及醋酸扭体法等作为基础研究方法,以高、中、低浓度益气活血方灌胃小鼠5天,研究该方的抗炎镇痛效果；3、60只SD大鼠,随机选取10只作为假手术组：仅暴露椎板,未造成脊髓压迫,以生理盐水(Normal Saline, NS)进行灌胃；其余的SD大鼠采用“自体第七颈椎棘突椎间移植法”制造大鼠脊髓型颈椎病模型,术后第1天根据脊髓功能的评分将其分为：A组：造模后,以高、中、低浓度益气活血方进行灌胃；B组：造模后,以颈复康进行灌胃；C组：造模后,以NS进行灌胃。造模后对大鼠进行为期4周的连续灌胃,每周均采用改进Rivaling的斜板试验、改良Tarlov评分、BBB评分等方法评估大鼠运动功能的恢复程度。灌胃结束后,采用HE染色法观察受损脊髓组织的形态学改变和ELISA方法测定大鼠受损节段中SOD、MDA、PGE2、PLA2的含量。结果： 1、小鼠的日最大给药量是临床成人日用量的315倍,相当于生药的892.5g·kg-1,在该剂量下未见明显毒性反应,7天内未出现动物死亡现象；2、发现高、中、低浓度的益气活血方均能显著抑制二甲苯致小鼠耳肿胀的作用,能减轻冰醋酸致小鼠扭体反应次数。3、行为学观察：高、中、低浓度益气活血方及颈复康均能显著增加Rivaling的斜板试验的平均度数、Tarlov评分、BBB评分,与NS组相比效果显著(P0.05)。4、HE染色：受损脊髓组织形态学的观察结果显示,中药高、中、低浓度组以及颈复康组的受损脊髓组织内均可见胶质瘢痕和空洞形成,且空洞总面积均明显小于NS组。5、ELISA方法：通过检测大鼠受损脊髓组织中的炎症因子发现,高、中浓度益气活血方及颈复康均能显著降低受损脊髓组织内PGE2的含量(P0.05)；高、中、低浓度益气活血方及颈复康均能显著减低受损脊髓组织内PLA2的含量(P0.05)；高、中、低浓度益气活血方及颈复康颗粒均能显著增加大鼠受损脊髓组织内SOD的含量(P0.05)；高、中浓度的益气活血方以及颈复康颗粒均能显著降低脊髓组织内的MDA含量(P0.05)；低浓度益气活血方亦可降低其组织内PGE2、 MDA含量,但与对照组相比,无显著性差异(P0.05)。结论：益气活血方是安全的,并具有明显的抗炎镇痛作用,且能通过提高受损脊髓组织细胞中氧自由基的清除率以及抑制受损脊髓组织的炎症反应来促进脊髓功能的恢复。
[Abstract]:Objective: to evaluate the safety of clinical experience prescription "Yiqi Huoxue recipe" by acute toxicity test, and to study the anti-inflammatory and analgesic effect of the prescription by auricle swelling test and acetic acid writhing method. The effect of the prescription on the recovery of injured spinal cord nerve function in rats was studied by using the rat model of cervical Spondylotic myelopathy, and the related mechanism of promoting the recovery of spinal cord function was clarified. Methods: 1Twenty ICR mice were randomly selected as control group and 15 as experimental group. To study the acute toxicity of Yiqi Huoxue prescription by the method of maximum dosage; (2) taking xylene induced mouse ear swelling and acetic acid writhing as basic research methods, the anti-inflammatory and analgesic effect of this prescription was studied by oral administration of high, middle and low concentration of Yiqi Huoxue recipe for 5 days in mice. (3) 60 SD rats were randomly selected as sham-operated group (n = 10): normal Saline (NSN) was perfused with normal saline to expose vertebral lamina without spinal cord compression. The other Sprague-Dawley rats were divided into two groups according to the score of spinal cord function on the first day after operation. The low concentration Yiqi Huoxue prescription was perfused by stomach; Group B: after the model was made, the rats were perfused with Jingfukang. Group C: after the model was made, the rats were perfused with NS for 4 weeks. The modified Rivaling slant test was used every week to improve the Tarlov score. The degree of recovery of motor function in rats was evaluated by BBB score. After gastric perfusion, morphological changes of injured spinal cord tissues were observed by HE staining method and SOD in injured segments of rats were measured by ELISA method. Fruit setting: 1. The maximum daily dose of mice was 315 times that of clinical adults, equivalent to 892.5 g 路kg-1 of crude drugs. There was no obvious toxic reaction at this dose and no animal death occurred within 7 days. 2, found that high, medium and low concentrations of Yiqi Huoxue prescription can significantly inhibit xylene induced mouse ear swelling, can reduce the number of acetic acid-induced mice writhing reaction times .3.Behavioral observation: high, medium. The low concentration of Yiqi Huoxue recipe and Jingfukang could significantly increase the mean degree of Rivaling inclined board test and the BBB score. Compared with NS group, the results of HE staining showed that the histomorphology of injured spinal cord was high and moderate. The formation of glial scar and cavity were found in the injured spinal cord tissue of low concentration group and Jingfukang group, and the total area of cavity was significantly smaller than that of NS group (.5%). ELISA method: by detecting inflammatory factors in injured spinal cord tissue of rats, high levels of inflammatory cytokines were found. The medium concentration of Yiqi Huoxue recipe and Jingfukang could significantly reduce the content of PGE2 in injured spinal cord tissue. High, medium and low concentrations of Yiqi Huoxue prescription and Jingfukang can significantly reduce the content of PLA2 in injured spinal cord tissue. High, medium and low concentrations of Yiqi Huoxue prescription and Jingfukang granule could significantly increase the content of SOD in injured spinal cord tissue of rats. High and medium concentration of Yiqi Huoxue recipe and Jingfukang granule could significantly reduce the content of MDA in spinal cord tissue. Low concentration Yiqi Huoxue prescription can also reduce the content of PGE2 and MDA in its tissue, but there is no significant difference compared with the control group. Conclusion: Yiqi Huoxue prescription is safe. It also has obvious anti-inflammatory and analgesic effect and can promote the recovery of spinal cord function by increasing the clearance rate of oxygen free radicals in the cells of injured spinal cord tissue and inhibiting the inflammatory response of injured spinal cord tissue.
【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】：2016
【分类号】：R274.9

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