脑梗死气虚血瘀证大鼠模型的建立与评价

发布时间：2018-07-05 01:11

本文选题：脑梗死 + 气虚血瘀　；参考：《北京中医药大学》2007年硕士论文

【摘要】： 证候是疾病发生发展不同阶段的病情动态时空变化的概括,是中医辨证论治的核心,证候研究是中医学术研究的中心。证候与疾病密不可分,研究证候离不开疾病,研究疾病也离不开证候,这是中医对疾病与证候关系的经典认识方法。脑梗死是临床常见疾病,气虚血瘀证可贯穿于该病全程。本课题以脑梗死气虚血瘀证为切入点,通过线栓法和力竭性游泳,把脑梗死模型与气虚血瘀证候模型复为一体,力求使气虚作为始动因素、主要因素而贯穿气虚血瘀模型的全过程,建立与临床原型相似的脑梗死气虚血瘀证的病证结合模型,比较观察模型动物气虚血瘀表征与生物学指标变化,进一步探索脑梗死气虚血瘀证模型动物的敏感表征与指标变化的关系,确立该模型的评价体系。 1.目的: 本实验拟以力竭性游泳复合线栓法复制脑梗死气虚血瘀证病证结合动物模型,探索病证结合动物模型的制作方法,通过对宏观表征、神经病学评分、脑组织病理变化及相关生化指标变化的观察,寻找表征与其他相关指标的关系,建立病证结合模型的评价体系。 2.方法: 健康雄性SD大鼠64只,体重(200±10)g,适应性喂养3天后,随机分为4组:正常组、单纯游泳组、单纯手术组、病证结合模型组,每组大鼠16只。正常组与单纯手术组正常喂养3周,不进行任何干预;单纯游泳组与病证结合模型组每天进行力竭性游泳训练,持续3周。3周后单纯手术组与病证结合模型组通过线栓法建立缺血2小时,再灌24小时的脑梗死模型。对比观察游泳与非游泳大鼠体重、进食量、活动量及手术死亡率;对比观察手术前后大鼠一般状态,术后6小时按照Zea-longa评分标准对大鼠进行神经症状体征评分;术后24小时:○1每组大鼠随机各取3只心脏灌注固定后取脑,再以4%的多聚甲醛固定1周后石蜡包埋制作患侧大脑冠状切片,进行常规HE染色后光镜观察脑组织形态学改变;○2剩余大鼠断头取脑,制成脑组织匀浆进行生化指标检测,包括:丙二醛(MDA)含量及超氧化物歧化酶(SOD)活性测定,一氧化氮(NO)含量及一氧化氮合酶(NOS)活性测定,乳酸(LD)含量及乳酸脱氢酶酶(LDH)活性测定。所得数据用Excel软件处理,应用SPSS11.0软件行单因素方差分析。 3.结果: 通过三周游泳训练,单纯游泳组及病证结合模型组大鼠与正常组及单纯手术组相比,饮食量减少、体重增长缓慢;倦怠嗜睡,喜欠伸,攻击性、对抗性行为减少或消失;舌质明显暗淡,舌下脉络增粗,瘀紫。线栓法造成脑梗死模型后,单纯手术组死亡率(13%)明显低于病证结合模型组死亡率(37%),神经症状体征较轻;再灌24小时后,前组大鼠恢复好,活动能力较强。两组舌质较正常组更加黯淡,舌下脉络有不同程度的瘀紫,增长,但复合模型组舌象变化较单纯手术组明显。正常组及单纯游泳组大鼠脑组织无缺血损伤改变,但单纯游泳组大鼠脑组织血管增生明显;单纯手术组及复合模型组大鼠脑组织结构出现缺血性形态改变,复合模型组大鼠脑组织损伤更为严重。复合模型组脑组织中SOD活性(81.58±24.47)和单纯手术组脑组织中SOD活性(61.41±20.74)较正常组(106.66±6.40)显著降低P0.05。复合模型组脑组织中脂质过氧化产物MDA的含量(1.26±0.19)和单纯手术组脑组织中脂质过氧化产物MDA含量(1.03±0.31)较正常组(0.0.64±0.07)明显增多(P0.05)。 4.结论: 4.1在单次和反复多次力竭性游泳,造成气虚血瘀的基础上再复合脑梗死制作病证结合动物模型,从动物生活习性改变、一般状态改变,神经功能改变等表征观察,以及相关的病生理指标、生化指标检测,一定程度上综合模拟出脑梗死气虚血瘀证候特点。 4.2从宏观表征和微观指标较为客观评价气虚血瘀状态和相关病生理基础变化,为建立病证结合的评价方法提供了依据。
[Abstract]:Syndrome is the generalization of the dynamic and spatio-temporal change of disease dynamics in different stages of disease and development. It is the core of TCM syndrome differentiation and treatment. Syndrome research is the center of TCM academic research. Syndrome and disease are inseparable, research syndrome can not be separated from disease, and the study of disease is also inseparable from syndrome. This is the classic method of understanding the relationship between disease and syndrome in Chinese medicine. Infarction is a common clinical disease. Qi deficiency and blood stasis syndrome can run through the whole course of the disease. This subject takes the syndrome of qi deficiency and blood stasis of cerebral infarction as the breakthrough point. The model of cerebral infarction and Qi deficiency and blood stasis syndrome are integrated into one, and the whole process of Qi deficiency and blood stasis model is made through the whole process of qi deficiency and blood stasis model. To establish the model of syndrome combination of cerebral infarction and blood stasis syndrome of cerebral infarction, the characteristics of qi deficiency and blood stasis in model animals and the changes of biological indexes were compared, and the relationship between the sensitive characterization and index changes of the model animals of cerebral infarction and blood stasis syndrome was further explored, and the evaluation system of the model was established.
1. purpose:
In this experiment, we should use the method of exhausting swimming compound thread to copy the animal model of the syndrome of cerebral infarction and blood stasis syndrome, explore the method of making the disease syndrome combined with animal model, through the observation of the macroscopic characterization, neurology score, the pathological changes of the brain tissue and the changes of the related biochemical indexes, looking for the relationship between the characterization and other related indexes, and establishing the disease. The evaluation system of the combination model.
The 2. method:
64 healthy male SD rats, body weight (200 + 10) g, 3 days after adaptive feeding, were randomly divided into 4 groups: normal group, simple swimming group, simple operation group, syndrome combination model group and 16 rats in each group. Normal group and simple operation group were fed for 3 weeks without any intervention; the simple swimming group and the combination model group were exhausted every day for exhaustive swimming. After 3 weeks of training, 3 weeks after the operation, the model group with the combination of the simple operation group and the disease syndrome group established the cerebral infarction model after 2 hours of ischemia and reperfusion for 24 hours by the thread thrombus method. The body weight, food intake, activity and operation mortality of the swimming and non swimming rats were compared and observed, and the general state of the rats before and after the operation was compared and the Zea-longa score standard was observed after 6 hours after the operation. Rats were given neurological signs and signs, 24 hours after the operation: 0. 1 rats in each group were randomly selected for 3 heart perfusion and then taken the brain, then 4% paraformaldehyde was fixed for 1 weeks to make paraffin embedded cerebral coronal section, and the conventional HE staining was used to observe the morphological changes of the brain tissue. 2 the remaining rats were broken head to take the brain to make brain tissue homogenization. The pulp was tested for biochemical indexes, including the determination of malondialdehyde (MDA) content and superoxide dismutase (SOD) activity, the determination of nitric oxide (NO) and nitric oxide synthase (NOS) activity, the content of lactic acid (LD) and the activity of lactate dehydrogenase (LDH). The obtained data were processed by Excel software, and the single factor variance analysis of SPSS11.0 software was applied.
3. results:
After three weeks of swimming training, the rats in the simple swimming group and the disease syndrome combined model group were compared with the normal group and the normal group and the simple operation group. The diet was reduced and the weight growth was slow; the burnout lethargy, yawning, aggressive, antagonistic behavior decreased or disappeared; the tongue quality was dull, the sublingual vein was thickened and the blood stasis. After the cerebral infarction model, the simple operation was caused by the thread thrombus method. The mortality of the group (13%) was significantly lower than that of the combination model group (37%), and the signs of neurologic symptoms were lighter. After 24 hours of reperfusion, the rats in the former group recovered well and had stronger activity. The two groups of tongue were more gloomy than the normal group, and the sublingual veins had different degrees of blood stasis and growth, but the changes of tongue image in the compound model group were more obvious than those in the simple operation group.
There was no ischemia injury in the brain tissue of the normal group and the rats in the simple swimming group, but the cerebral tissue of the rats in the simple group and the compound model group had the ischemic morphologic changes. The brain tissue injury in the compound model group was more serious. The activity of SOD in the compound model group was (81.58 + 24.47). The activity of SOD in the brain tissue of the operation group (61.41 + 20.74) was significantly lower than that of the normal group (106.66 + 6.40). The content of the lipid peroxidation product MDA in the brain tissue of the P0.05. complex group (1.26 + 0.19) and the lipid peroxidation product MDA content in the brain tissue of the simple operation group (1.03 + 0.31) were significantly increased (P0.05) compared with the normal group (0.0.64 + 0.07).
4. conclusion:
4.1 on the basis of single and repeated exhaustive swimming, causing Qi deficiency and blood stasis on the basis of complex cerebral infarction, the disease syndrome combined with animal model, from animal life habit change, general state change, nerve function change and other characterization observation, related diseases physiological index, biochemical index detection, to a certain extent, simulate the deficiency of cerebral infarction Qi deficiency. Characteristics of syndrome of blood stasis.
4.2 the changes in the state of qi deficiency and blood stasis and the physiological basis of related diseases were objectively evaluated from the macroscopic and microscopic indexes, which provided the basis for the establishment of the evaluation method of the combination of disease and syndrome.
【学位授予单位】：北京中医药大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R-332

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