膈下逐瘀汤及拆方对微循环障碍大鼠的影响和机制研究

发布时间：2018-08-15 14:23

【摘要】：目的:微循环障碍的病理表现存在于心脑血管疾病、糖尿病、肿瘤等诸多高危的疾病,微循环障碍与中医的血瘀证有着密切的联系,充分发挥中医药的优势,治疗微循环障碍疾病具有重要的意义。本文观察了膈下逐瘀汤及其拆方对与微循环障碍模型大鼠的疗效及其机制,为进一步研究膈下逐瘀汤的影响,为临床的用药提供理论依据。方法:运用高分子右旋糖酐尾静脉注射的方法制作大鼠微循环障碍模型。将50只Wistar大鼠随机分为5组,每组10只,分别为正常对照组(简称“正常组”)、模型对照组(简称“模型组”)、膈下逐瘀汤高剂量组(简称“高剂量组”)、膈下逐瘀汤低剂量组(简称“低剂量组”)、活血拆方组(简称“拆方组”)。模型组、高剂量组、低剂量组、拆方组分别以10%高分子右旋糖酐进行尾静脉注射,剂量10ml/Kg,隔天注射一次,连续注射10次。对照组隔天尾静脉注射0.9%生理盐水,剂量为10ml/Kg。给药组在造模基础上,灌胃给药,灌胃量为10ml/Kg,隔天灌胃一次,连续灌胃十次。其中,膈下逐瘀汤水煎液,低剂量组给药量为7.5g/Kg,高剂量组给药量为11.5 g/Kg;活血拆方组给于当归、桃仁、红花、川芎、元胡水煎液,每日用药剂量为3g/Kg。对照组、模型组给于纯净水灌胃,灌胃量为10ml/Kg,隔天一次,共10次。末次给药后,晚上动物禁食、不禁水,第2天进行实验检测。5%水合氯醛麻醉,首先进行腹腔肠系膜微循环观察(主要检测流速、流态、动静脉比等,观察有无血瘀改变;其次观察有无出血等),其后抽取腹腔静脉血,进行血液流变学、血管内皮活性因子的检测(NO、ET、VEGF、ES含量)等,探讨微循环障碍、血瘀等改变的内在机制。并取肝、肺等脏器组织进行肝、肺脏器组织病理检测,重点观察组织内微循环损伤的改变。结果:膈下逐瘀汤高、低剂量组对微循环障碍大鼠有较好的疗效,且活血拆方组效果不如全方组。(1)肺、肝组织微循环损伤的治疗效果:第一,肺组织损伤的治疗效果:给与膈下逐瘀汤后,高、低剂量组大鼠的间质瘀血、出血状况均明显减轻,炎细胞浸润明显减少,间隔增厚的程度减轻,尤其是膈下逐瘀汤高剂量组肺组织微循环损伤的改善效果更为明显,P0.05,结果具有统计学意义。拆方组的改善效果不如膈下逐瘀汤高、低剂量组,P0.05,结果具有统计学意义。其二,肝组织损伤的治疗效果:膈下逐瘀汤高、低剂量组肝细胞比较均匀,无变性、坏死,肝小叶结构较为清晰,肝脏纤维间隔恢复、红细胞聚集之血瘀状况得到了较好的改善,尤其是高剂量组的治疗效果更为明显。活血拆方组大鼠肝脏的微循环瘀血状态也得到了改善,但效果不及高、低剂量组。(2)肠系膜微循环损伤的疗效:膈下逐瘀汤高、低剂量组动、静脉流速明显上升,血液流态主要表现为线流,血流瘀滞状态得到改善。活血拆方组,血液流速升高,P0.05,结果具有统计学意义,肠系膜微循环状态得到相应的改善,但改善效果不如膈下逐瘀汤给药组。膈下逐瘀汤高剂量组和低剂量组大鼠肠系膜微动脉增粗,微静脉变细,DA/DV比值变大,P0.05,结果具有统计学意义,数据结果可显示肠系膜血管壁受损得到改善,DA/DV比值接近正常组,肠系膜微循微循环状况良好,且高剂量组的治疗效果优于低剂量组。活血拆方组对肠系膜动、静脉的病理状况也有一定的疗效,但治疗效果不如膈下逐瘀汤全方组,P0.05,结果具有统计学意义。高剂量组和低剂量组出血像素点减少,出血面积小,出血症状得到较好的改善。活血拆方组的治疗效果不如膈下逐瘀汤的高、低剂量组。(3)对血流变指标的改善:膈下逐瘀汤高、低剂量组的全血高、低切粘度值以及血浆粘度值、全血高、低切还原粘度值、细胞压积、红细胞刚性指数、红细胞聚集指数都有了明显的降低,红细胞变形指数(TK)出现了升高,P0.05,结果具有统计学意义,活血拆方组的治疗效果不如膈下逐瘀汤给药组。(4)对血管活性因子的疗效:膈下逐瘀汤高、低剂量组NO含量升高,ET含量降低,差异具有统计学意义(P0.05),结果可表明血瘀的病理状况明显的减轻,微循环也得到较好的改善,且膈下逐瘀汤高剂量组的改善效果优于低剂量组。活血拆方组肺组织NO含量略有升高,ET含量显著升高,NO与ET含量的比例严重失调,但差异不具有统计学意义。活血拆方组血浆中NO含量升高,ET含量有较大幅度的升高,且ET含量差异不具有统计学意义。膈下逐瘀汤高、低剂量组VEGF含量降低,ES含量升高,差异具有统计学意义(P0.05),结果可显示肺组织损伤得到修复,肺部微循环状况得到改善,且膈下逐瘀汤高剂量组的疗效优于低剂量组。拆方组与模型组比,拆方组VEGF含量显著降低,ES含量升高,差异具有统计学意义(P0.05),可见活血拆方组对肺部损伤有一定的治疗效果,但是治疗效果远不如膈下逐瘀汤高、低剂量组。大鼠血浆中VEGF和ES含量:从表中数据结果可以看出,正常组、模型组、膈下逐瘀汤高低及剂量组、活血拆方血浆中的VEGF含量极少,且结果差异不具有统计学意义。膈下逐瘀汤高、低剂量组大鼠血液中的ES含量减少,且高剂量组ES含量少于低剂量组,差异具有统计学意义(P0.05)。活血拆方组ES含量增多,差异具有统计学意义(P0.05)。结论:膈下逐瘀汤高、低剂量组对微循环障碍大鼠有较好的疗效,且活血拆方组效果不如全方组,膈下逐瘀汤活血方药与凉血止血药及行气药的配伍作用是重要的。
[Abstract]:Objective: The pathological manifestations of microcirculation disorders are found in many high-risk diseases such as cardiovascular and cerebrovascular diseases, diabetes mellitus, tumor and so on. The microcirculation disorders are closely related to the blood stasis syndrome of TCM. It is of great significance to give full play to the advantages of TCM in treating microcirculation disorders. Methods: The rat model of microcirculation disorder was established by injecting high molecular dextran into tail vein. Fifty Wistar rats were randomly divided into 5 groups, 10 rats in each group, which were normal control group. Normal group, model control group (model group), high-dose group of Gexia Zhuyu Decoction (high-dose group), low-dose group of Gexia Zhuyu Decoction (low-dose group), and Huoxue Jiefang group (split prescription group). The control group was given 0.9% normal saline by tail vein every other day, and the dosage was 10ml/Kg. On the basis of modeling, the administration group was given intragastric administration with the dosage of 10ml/Kg, once every other day, continuously for ten times. Among them, the decoction of Gexia Zhuyu decoction, the dosage of low-dose group was 7.5g/Kg, and the dosage of high-dose group was 7.5g/Kg. The dosage was 11.5 g/Kg; the decoction of Angelica sinensis, Peach kernel, Safflower, Ligusticum Chuanxiong and Corydalis Corydalis were given to the decoction of Huoxue decoction, and the daily dosage was 3 g/Kg. The control group was given pure water by gastric lavage, and the dosage was 10 ml/Kg every other day. The microcirculation of mesenteric cavity was observed (mainly to detect the flow velocity, flow pattern, arteriovenous ratio, etc.) and the blood stasis was observed; the abdominal venous blood was then collected for Hemorheology and the detection of vascular endothelial active factors (NO, ET, VEGF, ES content) to explore the internal mechanism of microcirculation disturbance and blood stasis. Results: The high and low dose of Gexia Zhuyu decoction had better curative effect on rats with microcirculation disturbance, and the effect of Huoxue Jiefang group was not as good as that of the whole prescription group. (1) The therapeutic effect of lung and liver microcirculation injury: First, the treatment of lung tissue injury. Results: After giving Gexia Zhuyu Decoction, the interstitial blood stasis and bleeding of rats in high and low dosage groups were alleviated, inflammatory cell infiltration and interval thickening were significantly reduced, especially in the high dosage group, the improvement effect of pulmonary microcirculation injury was more obvious, P 0.05, the results were statistically significant. Second, the treatment effect of liver tissue injury: high-dose Gexia Zhuyu decoction, low-dose group liver cells more uniform, no degeneration, necrosis, liver lobule structure is clearer, liver fibrous septum recovery, red blood cell aggregation of blood stasis has been better improved, especially (2) The effect of mesenteric microcirculation injury: the velocity of arteries and veins in the high and low doses of Gexia Zhuyu Decoction increased significantly, and the blood flow pattern was mainly linear and blood stasis state. The results showed that the mesenteric microcirculation was improved correspondingly, but the improvement effect was not as good as that of the Gexia Zhuyu Decoction group. The results showed that the damage of mesenteric vascular wall was improved, the DA/DV ratio was close to the normal group, the mesenteric microcirculation was good, and the effect of high dose group was better than that of low dose group. The treatment effect of Huoxue Jiefang group was not as good as that of the high and low dose groups of Gexia Zhuyu Decoction. (3) Improvement of hemorheological indexes: high Gexia Zhuyu Decoction, high whole blood, low shear viscosity of low dose group. The values of plasma viscosity, whole blood viscosity, low shear reduction viscosity, hematocrit, erythrocyte stiffness index, erythrocyte aggregation index were significantly reduced, erythrocyte deformability index (TK) increased, P 0.05, the results were statistically significant, the effect of Huoxue Jiefang group was not as good as that of Gexia Zhuyu Decoction. The difference was statistically significant (P 0.05). The results showed that the pathological condition of blood stasis was obviously alleviated, and microcirculation was also improved. The improvement effect of high dose group was better than that of low dose group. The content of NO in lung tissue of Huoxue Jiefang group was slightly increased. The content of NO and ET in plasma increased significantly in Huoxue Decoction group, and the content of ET was not statistically significant. The content of VEGF and ES in Gexia Zhuyu Decoction were decreased in high and low dose groups, and the difference was statistically significant. The results showed that lung tissue injury was repaired, the microcirculation of lung was improved, and the curative effect of high dose group was better than that of low dose group. The content of VEGF and ES in plasma of rats: From the data of table, we can see that the content of VEGF in plasma of normal group, model group, high and low Gexia Zhuyu Decoction and dose group is very little, and the difference is not statistically significant. The content of ES in blood of rats in dose group decreased, and the content of ES in high dose group was less than that in low dose group, the difference was statistically significant (P 0.05). It is important that the compatibility of the decoction for removing blood stasis and activating blood and cooling blood and hemostasis drugs and Qi compatibility.
【学位授予单位】：山东中医药大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R285.5

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