脑出血急性期中医分阶段治疗方案及蛭龙活血通瘀胶囊的干预作用机制研究

发布时间：2018-07-04 20:33

  本文选题：脑出血急性期 + 中医分阶段治疗　； 参考：《成都中医药大学》2016年博士论文

【摘要】：第一部分：临床观察目的：通过临床观察脑出血急性期应用中医分阶段治疗方案对患者相关指标的影响,研究中医分阶段治疗方案对脑出血急性期治疗的临床疗效,从而制定符合临床实际、规范化且行之有效、易于推广应用的脑出血急性期的临床治疗方案,以期提高该病的临床疗效。方法：采用随机平行对照的临床试验方法,纳入脑出血急性期患者,共60例,随机分为观察组和对照组,每组各30例,其中对照组采用西医常规治疗,包括稳定生命体征、控制血压、血糖、营养神经、改善脑代谢等；观察组在此基础上采用中医分阶段治疗方案,针对急性期不同阶段分别给予中药汤剂辨证施治,中成药、针灸理疗等方法进行治疗,两组均于治疗前及治疗后14d对患者的临床疗效及中医症状分级量表评分、NIHSS评分、Rankin评分、Barthel指数评分讲行检测,并于治疗前、治疗后7d及14天对患者颅内血肿的变化情况以及安全性指标,包括血常规、肝肾功、凝血等进行检测,从而比较两组的临床疗效和对上述指标的影响情况,分析并评价两组的有效性和安全性。结果：1、中医分阶段治疗方案组(观察组)患者的中医症状分级量表评分、NIHSS评分、Rankin评分、Barthel指数评分、颅内血肿体积改善情况均明显优于单纯西医治疗组(对照组),P0.05；观察组的中医证候疗效、西医临床疗效、颅内血肿疗效均明显高于对照组,P0.05；2、两种治疗方案对患者的生命体征、肝肾功、血常规、凝血等指标均未造成不良影响,具有较好的安全性。结论：对脑出血急性期患者采用中医分阶段治疗方案较单纯西医治疗能够更加有效的使患者神经功能得到改善,从而提高患者治愈率,并改善生活质量,安全有效,值得推广。第二部分：动物实验目的：探讨蛭龙活血通瘀胶囊对脑出血大鼠脑水肿、血脑屏障功能的影响,并研究该药脑保护的作用机制,为蛭龙活血通瘀胶囊在脑出血急性期的临床运用提供科学的实验依据。方法：取255只雄性、健康的SD大鼠,采用随机分组法分为假手术组、模型组、蛭龙活血通瘀胶囊高、中、低剂量组,各组又根据指标检测的时间不同划分为12h、48h、72h三个亚组,采用自体血注入法制备脑出血大鼠模型,各药物组于术后分别给予蛭龙活血通瘀胶囊灌胃,每日量分别为0.3g/kg、0.6g/kg和1.2g/kg,而模型组和假手术组则灌胃与药物组同等体积的生理盐水,3ml／次,给药次数及时间同药物组。各组进行相应干预后,在各规定的时间点对大鼠神经行为学评分(NSS)进行测定,取出脑组织,检测大鼠脑含水量及脑系数,HE染色对大鼠脑出血血肿周围组织行病理学观察,采用伊文思蓝法检测血脑屏障的通透性,电子显微镜观察血脑屏障的超微结构,Western-blot法检测与脑水肿相关的蛋白AQP-4、MMP-9及TIMP-1的表达,PT-PCR法检测AQP-4、MMP-9及TIMP-1 mRNA表达。结果：1、脑出血模型大鼠在术后12h可见脑组织水肿,48h及72h脑水肿程度逐渐加重,神经功能缺损情况逐渐明显,NSS评分随之升高,脑含水量及脑系数也逐渐升高,HE染色见脑组织明显水肿,间质疏松,血肿周围结构紊乱。蛭龙活血通瘀胶囊低、中、高剂量组在术后12h、48h及72h,大鼠脑水肿情况均较模型组有所改善,神经功能评分较之下降,脑含水量及脑系数也较之下降(P0.01),HE染色可见脑组织血肿周围水肿均有不同程度的改善,各药物组之间比较以高剂量组改善最为明显。2、脑出血模型大鼠在术后12h血脑屏障的通透性即可发现有明显改变,脑组织EB含量明显升高,术后48h时升高最为明显,72h时稍有下降。电镜超微结构显示术后48h,整个血管腔形态不规则,基膜厚薄不均,毛细血管周围水肿明显,组织稀疏,电子密度降低,内皮局部膨出,部分与基膜剥离,血脑屏障结构发生破坏。蛭龙活血通瘀胶囊低、中、高剂量组在术后12h、48h及72h,大鼠血脑屏障通透性较模型组均具有不同程度的改善(P0.01),脑组织EB含量较之有所下降,电镜超微结构观察发现血管周围水肿程度较为轻微,血脑屏障的结构得到明显改善,以高剂量组改善最为明显。3、脑出血模型大鼠在术后12h、48h及72h脑组织中AQP-4、MMP-9、TIMP-1蛋白及基因的表达均有明显上升,其中,AQP-4蛋白及基因表达呈进行性升高,在72h时升高最为明显,MMP-9、TIMP-1蛋白及基因表达也有明显升高,在48h时上升明显并达到高峰,在72h时略有下降。而蛭龙活血通瘀胶囊低、中、高剂量组在术后12h、48h及72h时大鼠脑组织AQP-4、MMP-9蛋白及基因的相对表达量均有不同程度的下降,TIMP-1蛋白及基因的相对表达量有所升高,与模型组比较,差异均具有显著性(P0.01),不同剂量组间比较,高剂量组最为明显(P0.01)。结论：1、蛭龙活血通瘀胶囊能够明显改善脑出血大鼠神经功能缺损症状,使神经功能缺损评分有所减轻,并降低脑出血大鼠脑含水量及脑系数,从而使脑组织水肿程度得到明显改善。2、蛭龙活血通瘀胶囊能够明显改善脑出血大鼠血脑屏障的通透性,使脑组织EB含量有所减轻,保护血脑屏障的组织结构避免血脑屏障受到破坏,从而减轻脑组织的水肿程度。3、蛭龙活血通瘀胶囊能够明显降低脑出血大鼠脑组织AQP-4、MMP-9蛋白及基因的表达,同时升高TIMP-1蛋白及基因的表达,从而改善血肿周围脑组织水肿。4、蛭龙活血通瘀胶囊上述药理作用存在一定的量效关系,以高剂量组作用最为明显。
[Abstract]:The first part: clinical observation: through the clinical observation of the acute phase of cerebral hemorrhage, the clinical effect of the phased therapy of TCM on the related indexes of the patients was studied, and the clinical effect of the phased therapy of TCM on the treatment of acute cerebral hemorrhage was studied so as to formulate the cerebral hemorrhage which was in accordance with the clinical practice, standardized and effective, and easy to be popularized and applied. In order to improve the clinical efficacy of the acute phase in order to improve the clinical efficacy of the disease. Methods: a randomized parallel controlled clinical trial was adopted to include 60 cases of acute cerebral hemorrhage. A total of 30 patients in each group were randomly divided into the observation group and the control group. The control group was treated with routine western medicine, including the stable vital signs, the control of blood pressure, blood sugar, and the camp. On the basis of this, the observation group was treated with Chinese traditional Chinese medicine decoction, Chinese traditional medicine, acupuncture and moxibustion and other methods for treatment. The two groups were treated before and after the treatment of the two groups, the clinical curative effect of the patients and the scale of TCM symptoms rating scale, NIHSS evaluation. Score, Rankin score, Barthel index score, and before treatment, after 7d and 14 days after treatment, the changes in the patient's intracranial hematoma and safety indexes, including blood routine, liver and kidney work, coagulation and so on, were detected to compare the clinical efficacy of the two groups and the effects on the above indications, and to analyze and evaluate the effectiveness and safety of the two groups. Results: 1, the TCM symptom classification scale score, NIHSS score, Rankin score, Barthel index score, the improvement of intracranial hematoma volume were obviously better than that of the simple western medicine treatment group (control group), P0.05, the curative effect of TCM syndrome in the observation group, the clinical effect of Western medicine and the curative effect of intracranial hematoma in the observation group. Obviously higher than the control group, P0.05; 2, two kinds of treatment schemes have no adverse effects on the patient's vital signs, liver and kidney work, blood routine, coagulation and so on. Conclusion: the treatment of acute cerebral hemorrhage in patients with acute stage of cerebral hemorrhage can be more effective than pure Western Medicine to make the patient's nerve function more effective. To improve the cure rate and improve the quality of life, and improve the quality of life, it is safe and effective, and it is worth popularizing. The second part: the objective of animal experiment: To explore the effect of hirudon Huoxue Tongyu Capsule on brain edema and blood brain barrier function of rats with cerebral hemorrhage, and to study the mechanism of the protective action of the drug for the acute stage of cerebral hemorrhage in the leeudon Huoxue Tongyu capsule. Methods: 255 male and healthy SD rats were divided into sham operation group, model group, hirudon Huoxue Tongyu capsule high, middle and low dose group, and three subgroups of 48h and 72h were divided into three subgroups according to the index detection time. The model of autologous blood injection was used to prepare the rat model of cerebral hemorrhage. After the operation, each drug group was given the gavage of hirudon Huoxue Tongyu capsule, respectively, 0.3g/kg, 0.6g/kg and 1.2g/kg, respectively, and the model group and the sham operation group were treated with the same volume of saline, 3ml / times, the times and time of the administration. The behavior score (NSS) was measured, the brain tissue was taken out, the brain water content and the brain coefficient were detected. HE staining was used to observe the tissue around the hematoma and hematoma in rats. The permeability of the blood brain barrier was detected by Evans blue, the ultrastructure of the blood brain barrier was observed by electron microscope, and the protein A related to brain edema was detected by the Western-blot method. The expression of QP-4, MMP-9 and TIMP-1 and the expression of AQP-4, MMP-9 and TIMP-1 mRNA were detected by PT-PCR. Results: 1, the brain edema was observed at 12h in the rat model of cerebral hemorrhage, the degree of 48h and 72h cerebral edema gradually increased, the nerve function defect was gradually obvious, the NSS score increased, the brain water content and brain coefficient increased gradually, and the brain tissue was stained to see the brain tissue. Edema, interstitial loosening, structure disorder around hematoma, leech Huoxue Tongyu capsule low, middle, high dose group after 12h, 48h and 72h, brain edema in rats were improved compared with the model group, neural function score was lower, brain water content and brain coefficient decreased (P0.01), HE staining showed that edema around brain hematoma around hematoma can be found to be different The improvement of the degree was the most obvious.2 in the high dose group. The permeability of the 12h blood barrier in the cerebral hemorrhage model rats could be found obviously, the EB content of the brain tissue increased obviously, the 48h increased most obviously after the operation, and the 72h decreased slightly. The ultrastructure of the electric microscope showed 48h after operation, and the whole vascular cavity form was not. Rules, the thickness of the basement membrane was uneven, the edema around capillaries was obvious, the tissue was sparse, the electron density was reduced, the endothelia was partially expanded, the blood brain barrier structure was partially destroyed by the basement membrane and the blood brain barrier structure was destroyed. The permeability of the blood brain barrier in the high dose group was 12h, 48h and 72h in the high dose group, and the permeability of blood brain barrier in the rat was improved to a different degree (P 0.01) the content of EB in brain tissue was lower than that of it. Ultrastructural observation of electron microscope showed that the degree of edema around blood vessels was slight, the structure of blood brain barrier was obviously improved, and the most obvious.3 was improved in high dose group. The expression of AQP-4, MMP-9, TIMP-1 protein and gene expression in 12h, 48h and 72h brain tissues of cerebral hemorrhage model rats were obvious. In addition, the expression of AQP-4 protein and gene showed progressive increase, the most obvious increase at 72h, MMP-9, TIMP-1 protein and gene expression was also significantly increased, at the time of 48h increased and reached the peak, a slight decrease in 72h, while the leech dragon Huoxue Tongyu capsule was low, medium and high dose group in 12h, 48h and 72h rat brain AQP-4, MMP-9 protein in postoperative 12h, 48h and 72h. The relative expression of the gene and the relative expression of TIMP-1 protein and gene increased. Compared with the model group, the difference was significant (P0.01). The high dose group was the most obvious (P0.01). Conclusion: 1, hirudon live blood Tongyu capsule can obviously improve the neural function deficiency in rats with cerebral hemorrhage. The loss of symptoms, reduce the score of neural function defect, reduce the brain water content and brain coefficient of cerebral hemorrhage rats, so that the degree of brain edema is obviously improved by.2. The permeability of blood brain barrier in rats with cerebral hemorrhage can be improved obviously, the content of EB in brain tissue is reduced and the tissue structure of the blood brain barrier can be protected. To avoid the damage of blood brain barrier and reduce the edema degree of brain tissue.3, hirudon Huoxue Tongyu capsule can obviously reduce the expression of AQP-4, MMP-9 protein and gene in brain tissue of rats with cerebral hemorrhage, and increase the expression of TIMP-1 protein and gene, so as to improve the pharmacological action of the edema.4 of the brain group around the hematoma and the capsule of hirudon Huoxue Tongyu capsule. There is a certain dose effect relationship. The effect of high dose group is the most obvious.
【学位授予单位】：成都中医药大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R277.7
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本文编号：2097338