【作者】 乐薇；

【导师】 陈邦国；

【作者基本信息】 湖北中医药大学 ， 针灸推拿学， 2013， 博士

【摘要】 目的高脂血症是指机体内脂肪代谢或转运异常所致。大量的流行病学调查和研究均表明,高脂血症是动脉粥样硬化、高血压、冠心病、糖尿病等多种疾病发生的重要危险因素。近几年非药物疗法治疗高脂血症受到重视,尤其是临床应用针刺疗法治疗高脂血症,有了较大的进展,疗效也较为显著,显示出独特的优势。本实验旨在以高脂血症大鼠模型为研究对象,以胆固醇逆转运通路为切入点,观察各组大鼠血脂水平、血液流变学、巨噬细胞炎症因子、肝脏和主动脉弓组织形态学变化及ABCA1、SR-BI、PPAR Y、IGF-1的基因、蛋白表达,探讨电针丰隆穴对胆固醇逆转运通路的影响以及防治高脂血症的作用机制。方法选取成年雄性大鼠40只,适应性喂养1周后,将实验大鼠随机分空白对照组、模型+普通饲料组、模型+高脂饲料电针组、模型+普通饲料电针组这4组,每组10只。治疗结束后,将各组大鼠经颈动脉取血,测定血脂(TC、TG、HDL-C、LDL-C)含量及血液流变学指标(全血高切相对指数、全血低切相对指数、血浆黏度、红细胞聚集指数)。然后分离各组大鼠腹腔巨噬细胞,加荧光标记的炎症因子分子抗体后应用流式细胞仪检测ICAM-1、MCP-1、TNF-α、IL-6的含量。用HE染色法,在光镜下观察各组大鼠肝脏组织和主动脉弓组织形态学变化情况。检测肝脏组织ABCA1、 SR-BI、PPARγ、IGF-1的基因(mRNA)表达影响和蛋白含量。结果①与空白对照组相比较,其余三组血清TC值均显著上升,有非常显著性差异(P<0.01)。模型+普通饲料组和模型+高脂饲料电针组与空白对照组比较血清LDL-C值显著上升,差异具有非常显著性(P<0.01),而HDL-C值明显下降,差异具有显著性(P<0.05)。与模型+普通饲料组相比较,模型+高脂饲料电针组和模型+普通饲料电针组血清TC、LDL-C值均显著降低,有非常显著性差异(P<0.01)。②除模型+普通饲料电针组外,其余各组大鼠血液流变学(全血高切相对指数、全血低切相对指数、血浆黏度、红细胞聚集指数)与空白对照组比较均有所增加,具有显著型差异(P<0.05,或P<0.01)；与模型+普通饲料组比较,模型+普通饲料电针组大鼠血液流变学各项指标均有明显下降,有非常显著性差异(P<0.01)。③除模型+普通饲料电针组组外,其余各组大鼠腹腔巨噬细胞内ICAM-1、 MCP-1、TNF-a、IL-6与空白对照组比较均有明显升高(P<0.01,P<0.05)。与模型+普通饲料组比较,模型+普通饲料电针组的IL-6有明显减少(P<0.05), ICAM-1、MCP-1、TNF-a均显著减少,两组比较具有非常显著性差异(P<0.01)。④模型+普通饲料组大鼠肝脏组织可见中度到重度弥漫性脂肪变性,肝脏组织已经出现损害,有脂肪沉积情况。主动脉弓动脉管壁厚薄不均,内膜增厚和脂质沉着,主动脉内皮细胞严重破坏,内膜不连续,大量的泡沫细胞沉积于内膜下层,可见病变脂纹脂斑。而高脂造模成功后给予针刺治疗同时分别加高脂饲料、普通始料喂养的两组情况有不同程度改善,肝脏组织病变程度有所减轻,肝细胞结构部分修复,脂肪沉积减轻,主动脉弓组织血管壁结构有所修复清晰,内皮下脂质及炎性细胞浸润减少,中膜弹力膜有所修复。⑤与空白对照组比较,除了模型+普通饲料电针组外,其余两组ABCA1、SR-BI、PPARγ均有显著降低(P<0.01),IGF-1有显著升高(P<0.01)；与模型+普通饲料组比较,模型+高脂饲料电针组只有SR-BI和IGF-1这两个数值有差异性(P<0.05),而模型+普通饲料电针组的ABCA1、SR-BI、PPARγ均有显著降低(P<0.01), IGF-1有显著升高(P<0.01)；与模型+高脂饲料电针组比较,模型+普通饲料电针组的ABCA1、SR-BI、PPAR γ、IGF-1均有显著差异性(P<0.05,P<0.01)。结论电针丰隆穴能降低高脂血症大鼠TC、LDL-C水平,升高HDL-C水平,改善高脂状态下血液浓、黏、聚、凝的高危险状态,减少大鼠巨噬细胞内炎症因子ICAM、MCP-1、TNF-a、IL-6的含量,改善大鼠肝脏及主动脉弓形态学变化,有效抑制脂肪变性的发生程度,清除脂肪细胞,保护血管内皮,减少脂质沉积,对于动脉硬化早期的血管壁具有一定的保护作用。电针丰隆穴通过提高胆固醇逆转运通路两个重要功能蛋白ABCA1、SR-BI的表达,加速脂质代谢,达到治疗高脂血症的目的。更多还原

【Abstract】 Objective:Hyperlipidemia refers to the abnormality in the body fat metabolism or transporter. A large number of epidemiological surveysthat hyperlipidemia is an important risk factor for atherosclerosis, high blood pressure, coronary artery disease. In recent years, non-drug therapy for hyperlipidemia attention, especially the clinical appl i cat ion of acupuncture in the treatment of hyperlipidemia, has made significant progress, the effects are more significant, showing a unique advantage. The purpose of this experiment is to hyperlipidemia rat model for the study of reverse cholesterol transport pathway as an entry point to observe the rats in each group lipid levels, hemorheology, macrophage inflammatory factor, liver and aortic arch tissue morphology. changes ABCA1, SR-BI, PPARγ, IGF-1gene and protein expression, electro-acupuncture the Fenglong on reverse cholesterol transport pathway the prevention and control of the mechanism of action of hyperlipidemia.Methods:Select40adult male SD rats, after adaptive feeding for1week, the rats were randomly divided into blank control group, model+normal diet group, model+high fat diet EA group, model+normal diet EA group. Each one has ten rats. After the end of treatment, the rats in each group through carotid artery blood, serum lipids (TC, TG, HDL-C, LDL-C) content and blood rheology indicators (high shear blood relative index, whole blood low shear relative index, plasma viscosity, erythrocyte aggregation index). Peritoneal macrophages of rats in each group were then separated and analyzed by flow cytometry plus fluorescently labeled antibody inflammatory cytokines molecules ICAM-1, MCP-1, TNF-α, IL-6levels. HE staining, is aim to observe the tissue morphological changes of the the rats liver tissue and the aortic arch in the light microscope. Detection of liver tissue of ABCA1and SR-BI, PPAR y, IGF-1gene (mRNA) expression and protein content.Results:①The with blank control group relative comparison, the remaining three groups plasma TC values are significant with the rise, there are very significant with difference (P<0.01), model+ordinary feed group and model+high-fat feed electric acupuncture group plasma LDL-C values significantly with the rise, the difference has very significantly with sex (P<0.01), HDL-C values significantly decreased, significant sex (P<0.05) difference; model+ordinary feed group phase comparison model+high-fat feed EA group and model+normal diet EA plasma TC, LDL-C values were significantly reduced, and there was a significant difference (P<0.01).(2) In addition to the model+EA group beginning material remaining rats hemorheology (whole blood high shear relative index of whole blood relative index, plasma viscosity, erythrocyte aggregation index) compared with the blank control group, both an increase in type differences (P<0.05, P<0.01); model+common starting material group Hemorheology model+normal diet EA group was significantly decreased (P<0.01).③In addition to the model+normal diet EA group group, the rest of the rats in each group peritoneal macrophages ICAM-1, MCP-1, TNF-a, IL-6compared with the blank control group were significantly increased (P<0.01, P<0.05). Compared with the model+ordinary feed group model+feed electric needle group in addition of IL-6have decreased significantly (P <0.05), the expression of ICAM-1, MCP-1, of TNF-a were very significantly with the decline, with a significant differences (P <0.01).④model+normal diet group rat liver tissue visible moderate to severe diffuse fatty degeneration of the liver tissue damage and fat deposition. The arterial wall of the aortic arch uneven thickness, intimal thickening and lipid calm, severely damaged aortic endothelial cells, endometrial discontinuous foam cells are deposited in the intima lower spot visible lesions fatty streaks fat. Given acupuncture treatment of hyperlipidemia successful modeling and high fat diet, the ordinary start feeding feeding two sets of circumstances are different degrees of improvement, the severity of liver tissue has been reduced, the structural part of the repair of liver cells, reduce fat deposition, aortic arch organizations vessel wall structure was repaired clear, reduce of subendothelial lipid and inflammatory cell infiltration, membrane stretch film some repair.⑤with blank control group, model+normal diet EA group, and the remaining two groups of ABCA1and SR-BI of PPARγ were significantly lower (P<0.01), and IGF-1significantly increased (P<0.01); model+normal diet group compared with the model+high fat diet EA group only SR-BI and IGF-1two numerical differences (P<0.05), while the model+normal diet EA group ABCA1, SR-BI, PPAR y were significantly (P<0.01) reduction in IGF-1significantly increased (P<0.01); model+high fat diet EA group, model+normal diet EA group ABCA1, SR-BI, PPAR Y, IGF-1were significant differences (P<0.05, P<0.01).Conclusion:EA Fenglong reduce hyperlipidemia rats TC, LDL-C levels, increased HDL-C levels, improve blood concentration in high-fat state, sticky, poly, condensate at high risk status, reduce macrophage inflammatory cytokines of ICAM, MCP-1, TNF-a and IL-6, improve rat liver and aortic arch morphological changes effectively suppress the occurrence of steatosis remove fat cells protect vascular endothelial reduce fatsedimentary, early atherosclerosis vascular wall has a protective effect. Electro-acupuncture the Fenglong by elevate cholesterol reverse transport the path two important functional protein of ABCA1and SR-BI expression, accelerated lipid metabolism, to achieve the purpose of the treatment of hyperlipidemia.更多还原