【作者】 沈诗军；

【导师】 周定刚；

【作者基本信息】 四川农业大学 ， 基础兽医学， 2008， 硕士

【摘要】 走马胎为紫金牛科紫金牛属植物,全株具有活血化瘀等功效,为民间常用的跌打伤科药。目前该科属植物的研究报道很少,有关走马胎对动物体内血栓形成、脂质代谢及微循环等方面的研究国内外尚未见报道。据此,本文实施以下相关试验:试验一观察走马胎提取液对病理模型SD大鼠体内凝血系统、血液流变学、脂质代谢和肺组织损伤的影响。将36只SD大鼠随机分为6组,即病理模型组、生理盐水组、PNS药物组和走马胎提取液高中低剂量组（分别为10 g生药/kg、5 g生药/kg、2.5 g生药/kg）。按0.08 mg/kg WB皮下注射肾上腺素,每日1次,连续12 d复制动物病理模型,然后灌胃给药,每日1次,连续12 d治疗后,测定SD大鼠血液中生理生化指标及光镜下观察肺组织病理变化。结果表明,与生理盐水对照组相比,病理模型组SD大鼠体内PT、APTT显著缩短（P＜0.05）,全血低切率（6和12 r/min）黏度、血浆Fg和MDA含量显著升高（P＜0.01,P＜0.05）,CAT和SOD活性显著降低（P＜0.05）,光镜观察显示,肾上腺素诱导的病理模型组肺泡隔和肺泡腔内可见大量血细胞漏出,肺泡上皮细胞坏死、脱落,肺泡隔明显增厚,部分肺泡塌陷、不张;与病理模型组相比,高中剂量组全血低中切率（6、12和30 r/min）黏度显著降低（P＜0.01,P＜0.05）,高剂量组SD大鼠体内PT、TT和APTT均显著延长（P＜0.05）,血浆Fg、MDA和TG含量显著降低（P＜0.01,P＜0.05）,CAT和SOD活性显著升高（P＜0.01,P＜0.05）,中剂量组NO含量显著上升（P＜0.05）,高中剂量组肺泡结构损伤轻微,肺泡隔厚度明显降低,肺泡腔内清晰,壁较光滑。试验二观察走马胎提取液对病理模型家兔机体内血栓形成、凝血系统、脂质代谢和血液流变学的影响。将48只家兔随机分为病理模型组、生理盐水对照组、PNS药物组、同时设走马胎提取液高中低剂量组（分别为15 g生药/kg、10 g生药/kg、5g生药/kg）,每组各8只。除生理盐水对照组外,将其余各组家兔分别沿其耳缘静脉缓慢注入BSA,每间隔3 d注射一次,每次125 mg/kg WB,共4次。每次注射BSA间隔期的前两天,每天皮下注射肾上腺素一次,每次0.1 mg/kg,WB共8次,复制动物病理模型共16 d。然后灌胃给药,每日1次,连续10 d治疗后,测定家兔血液中生理生化指标。结果表明,与生理盐水对照组相比,病理模型组家兔体内PT、APTT显著缩短（P＜0.01,P＜0.05）,Fg、MDA和PMP含量显著升高（P＜0.01,P 0.05）,全血切率黏度也有所升高（P＞0.05）,血浆中NO含量、SOD活性显著降低（P＜0.05）,血液中MCV、HCT、MPV、MCHC显著升高（P＜0.01,P＜0.05）,FV:CA、FⅦ:CA显著升高（P＜0.01,P＜0.05）;与病理模型组相比,高剂量组家兔体内全血低切率（6 r/min）黏度和血浆Fg、MDA和PMP含量显著降低（P＜0.01,P＜0.05）,PT、TT和APTT均极显著延长（P＜0.01）,CAT和SOD活性显著升高（P＜0.05）,血液中MCV、HCT显著降低（P＜0.05）,FⅤ:CA、FⅦ:CA极显著降低（P＜0.01）。试验三研究走马胎提取液对SD大鼠肠系膜微循环的作用。利用10^-8 mol/L NA复制微循环障碍,并于滴加NA前及滴加NA后5、10、15、20 min分别观察记录微循环各指标变化。结果表明,与生理盐水对照组相比,病理模型组SD大鼠滴加NA后,在5、10 min时微动脉和微静脉血管内径显著缩小（P＜0.01,P＜0.05）,在5、10、15、20 min时毛细血管内径和毛细血管开放数都极显著减少（P＜0.01）,在5、10、20 min时微循环血流速和血流态显著减慢（P＜0.01,P＜0.05）;与病理模型组相比,高剂量组SD大鼠在滴加NA后微血管内径均有所扩大（P＞0.05）,尤其是毛细血管内径在10、15、20 min时扩大尤为显著（P＜0.01,P＜0.05）,而且高剂量组还能增加毛细血管开放数,加快红细胞流通和改善红细胞流态,随时间延长,作用逐渐明显,在20 min时毛细血管开放数、红细胞血流速和血流态均有显著性差异（P＜0.01,P＜0.05）。综上所述,走马胎提取液通过延长动物体内PT、TT和APTT,降低FⅤ:CA、FⅦ:CA、全血黏度及血浆Fg含量,抑制机体内、外源性凝血过程,从而阻止血栓形成和减轻肺组织损伤;走马胎提取液能够降低体内MDA含量、升高NO含量、增强CAT和SOD活性,抑制脂质过氧化,发挥抗氧化作用,从而稳定血管内皮细胞和调节脂质代谢;走马胎提取液还能扩张毛细血管,增加毛细血管开放数量,加快红细胞流速和流态,改善机体微循环。更多还原

【Abstract】 Ardisia gigantifolia Stapf of Myrsinaceae genera, which is one of traditional Chinese herbal medicine, plays a role in invigorating blood circulation, eliminating stasis, etc. And it used to treat the wounded after a fall in the area of Yao minority in Southern China commonly. At present, experimental studies of Ardisia gigantifolia Stapf are rarely reported, especially, effects on thrombosis, dyslipidoses, microcirculation etc are not yet reported in domestic and abroad. Therefore, according to studies on effects of therapy with Chinese herbal medicine on thrombosis, dyslipidoses and microcirculation in domestic and abroad, three systemical trials were done.Trial 1 investigated effects of Ardisia gigantifolia Stapf extract on blood clotting system, lipid metabolism, hemorrheology and pulmonary injury in rat. 36 Sprague-Dawley （SD） rats were randomly divided into 6 groups（n=6 in each group）, including the control, patho-model, PNS and 3 Ardisia gigantifolia Stapf -treated model groups of 10 g/kg, 5 g/kg, 2.5 g/kg crude drug. Except for the control group that was injected with 0.9%NaCl solution, all others were subcutaneously injected with adrenailne 0.08 mg/kg body weight （WB） once a day for 12 days to induce blood stasis and pulmonary injury. After patho-model was established, panax notoginseng saponins（PNS） and 3 Ardisia gigantifolia Stapf -treated model groups were intragastrically treated once a day for 12 days. Whereas the control and patho-model groups were administered with the same volume of 0.9% NaCl solution in the same way. After the model-treatment groups were treated, blood physiological and biochemical index were assayed, and lungs were removed en bloc and treated by 10% formalin for pathological test. This results showed that there was a significant change in prothrombin time（PT）, activated partial thromboplastin time（APTT）, blood 6 and 12 r/min shear rate viscosity, levels of fibrinogen（Fg） and malonaldehyde（MDA）, activities of catalase（CAT） and superoxide dismutase（SOD）, and alveolar septa width between the control and the patho-model groups （P<0.01, P<0.05）. In addition, light microscopic viewing of the slides demonstrated a marked pulmonary injury in the patho-model group. When compared with the patho-model group, 10 and 5 g/kg groups significantly decreased blood 6, 12 and 30 r/min shear rate viscosity（P<0.01, P<0.05）. In the model-treated group of 10 g/kg, a significant prolonged PT, thrombin time（TT） and APTT, and decrease in levels of MDA, Fg and triglyceride（TG）, and also increase in activities of CAT and SOD were seen （P<0.01, P<0.05）. The model-treated group of 10 g/kg obviously increased level of nitrogen monoxidum（NO） （P<0.05）. Furthermore, HE staining showed that pulmonary injury was obviously improved in the 10 and 5 g/kg groups.Trial 2 investigated effects of Ardisia gigantifolia Stapf extract on blood clotting system, lipid metabolism and hemorrheology in rabbit. 48 rabbits were divided into 6 groups randomly （n=8 in each group）, including the control, patho-model, PNS and 3 Ardisia gigantifolia Stapf extract-treated model groups of 15 g/kg, 10 g/kg, 5 g/kg crude drug. All groups, except for the control group that was injected with 0.9%NaCl solution, others were slowly injected intravenously with bovine serum albumin（BSA） （125 mg/kg WB） at 1, 5, 9, 13 days, and adrenaline was subcutaneously injected at 2, 3, 6, 7, 10,11, 14, 15 days, respectively to induce vascular endothelial cell （VEC） injury and a disorder of lipid metabolism according to a technique used by Van Winkle and Levy. After patho-model establishment, PNS and 3 Ardisia gigantifolia Stapf extract-treated model groups were intragastrically treated once a day for 10 days, whereas the control and patho-model group were administered with the same volume of 0.9% NaCl solution in the same way. Blood physiological and biochemical index were assayed. This results showed that there was a significant change in PT, APTT, levels of Fg, NO, MDA and SOD, platelets （PLT）, mean cell volume （MCV）, mean platelet volume（MPV）, activities of coagulation factor V（FV:CA） and coagulation factorⅦ（FⅦ:CA） between the control and the patho-model groups（P<0.01, P<0.05）. When compared with the model group, the results also showed that 15 g/kg group significantly prolonged PT, TT and APTT （P<0.01）, increased activities of CAT and SOD（P<0.05）, and decreased levels of Fg, MDA and PMP, MCV and hematocrit（HCT） （P<0.01, P<0.05）. In addition, 15 g/kg dose group decreased FⅤ:CA and FⅦ:CA significantly（P<0.01）. Trial 3 investigated effects of Ardisia gigantifolia Stapf extract on the mesenteric microcirculation of SD rat. SD rats were dripped with noradrenaline（NA） of 10^-8 mol/L to induce disturbance of microcirculation, and effects of Ardisia gigantifolia Stapf extract on the changes of the mesenteric microcirculation of rat were observed before and after adrenaline was dripped. This results showed that there was a significant change in internal diameter of mesenteric arteriole, veinule and blood capillary, and mesenteric capillary count, mesenteric capillary blood flow and blood fluid state between the control and the patho-model groups（P<0.01, P<0.05）. When compared with the patho-model group, It was found that Ardisia gigantifolia Stapf of 10 g/kg crude drug could improve internal diameter of mesenteric micrangium, especially internal diameter of mesenteric blood capillary （P<0.01, P<0.05）. In addition, Ardisia gigantifolia Stapf of 10 g/kg crude drug could significantly improve mesenteric capillary count, capillary blood flow and blood fluid state（P<0.01, P<0.05）.In short, Ardisia gigantifolia Stapf extract could significantly prolong PT, TT and APTT, decrease FⅤ:CA, FⅦ:CA, blood shear rate viscosity and level of Fg in vivo to inhibit intrinsic and extrinsic coagulation system, thus to prevent thrombogenesis and attenuate pulmonary tissue damage. Ardisia gigantifolia Stapf extract could obviously decrease MDA level, increase NO level, activities of CAT and SOD to stop lipid peroxidation and play an important role in antioxidation, thus to stabilize VEC and regulate dyslipidoses. In addition, Ardisia gigantifolia Stapf extract could significantly improve mesenteric micrangium, capillary count, capillary blood flow and blood fluid state to be effective in the treatment of disturbance of microcirculation.更多还原