北五味子醇提物抗动脉粥样硬化作用及其机制研究

发布时间：2018-02-25 00:18

本文关键词： 北五味子醇提物动脉粥样硬化氧化应激内皮蛋白表达　出处：《西南交通大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的对北五味子醇提物抗动脉粥样硬化的作用进行研究,并探讨其作用机制。方法采用腹腔注射VD3+高脂饲料喂养9周建立大鼠动脉粥样硬化模型,以辛伐他汀作为阳性对照(4mg/kg/d),灌胃北五味子低(0.35g/kg/d)、中(0.7g/kg/d)、高(1.4 g/kg/d)剂量醇提物3周。双试剂直接法测定大鼠血清中甘油三酯(TG)、低密度脂蛋白(LDL-C)、高密度脂蛋白(HDL-C)的含量,微板法测定血清中谷丙转氨酶(ALT)、谷草转氨酶(AST)的活性,对肝组织、主动脉进行苏木精-伊红(HE)染色,观察各组大鼠肝脏以及主动脉的病理改变。生化法检测大鼠血清中谷胱甘肽-过氧化物酶(GSH-PX)、过氧化氢酶(CAT)、超氧化物歧化酶(SOD)的活性以及丙二醛(MDA)的含量;利用免疫组织化学法检测大鼠主动脉核因子相关因子-2(Nrf-2)、血红素氧合酶(HO-1)蛋白的表达情况,采用酶联免疫吸附(ELISA)法检测血清中氧化低密度脂蛋白(ox-LDL)、内皮素-1(ET-1)、血栓素B2(TXB2)、6-酮-前列腺素F1α(6-keto-PGF1α)的表达情况,以说明北五味子醇提物治疗动脉粥样硬化的作用机制。结果大鼠动脉粥样硬化模型成功建立,模型组大鼠主动脉可见明显的斑块形成,血管壁内膜增厚,斑块内可见数量不等的泡沫细胞。与模型组对比,北五味子醇提物各组的主动脉可见内膜、中膜和外膜,结构相对完整,斑块形成不明显,血管内膜增厚,平滑肌纤维病变较轻,可非常显著降低AS模型大鼠主动脉斑块面积占血管壁面积的百分比(P0.01或P0.001),还可显著降低主动脉斑块中的泡沫细胞数量(P0.05或P0.01)。肝组织病理观察结果显示,模型组肝脏中肝细胞胞浆内可见较多大小不一的空泡,高剂量组与模型对照组相比胞质内未见明显空泡且无病变。辛伐他汀组及北五味子醇提物各剂量组与模型对照组比较,能极显著降低大鼠血清中TG的含量(P0.001),显著性降低LDL-C的含量(P0.001或P0.01或P0.05),中、高剂量组可显著性升高HDL-C的含量(P0.01或P0.05);辛伐他汀组及北五味子醇提物各组可显著性降低大鼠血清ALT、AST的活性(P0.001或P0.01或P0.05)。北五味子醇提物组各组能显著增强血清GSH-PX的活性,极显著性降低血清中MDA的含量(P0.001),低、中剂量组可极显著性增强CAT的活性(P0.001),低剂量组可显著增强SOD的活性(P0.01);进一步的作用机制研究结果显示,北五味子醇提物各组均能极显著性上调大鼠主动脉Nrf-2的蛋白表达(P0.001),高剂量组能显著性上调HO-1的蛋白表达(P0.01)。北五味子醇提物各组均能显著降低ox-LDL的表达(P0.01),极显著性上调6-keto-PGF1α的表达(P0.001),低、中剂量组可显著降低ET-1的表达(P0.01或P0.05),而各剂量组对TXB2的表达无显著影响。结论北五味子醇提物具有明显的抗动脉粥样硬化作用,可促进大鼠血脂代谢,其机制可能与激活Nrf-2/ARE通路,抑制ox-LDL、ET-1的蛋白表达,调控血浆中TXB2和6-keto-PGF1α的平衡,调节机体氧化应激水平并维持内皮细胞功能有关。
[Abstract]:The purpose of anti atherosclerosis extract of Fructus schisandrae chinensis alcohol and the role of research, and to explore its mechanism. Methods for 9 weeks to establish the rat model of atherosclerosis by intraperitoneal injection of VD3+ high fat diet with simvastatin as positive control (4mg/kg/d), intragastric administration of five kinds of North Zi Di (0.35g/kg/d), in (0.7g/kg/d), high (1.4 g/kg/d) dose of ethanol extract for 3 weeks. Determination of triglycerides in serum of rats with double reagent direct method (TG), low density lipoprotein (LDL-C), high density lipoprotein (HDL-C) content, micro plate method for the determination of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) activity of liver tissue and aorta were stained with hematoxylin eosin (HE) staining, observe the change of liver and aortic pathology. Glutathione peroxidase were detected in rat serum (GSH-PX), catalase (CAT), superoxide dismutase (SOD) activity and C Two aldehyde (MDA) content; detection of nuclear factor of aortic related factors in rats using immunohistochemical method, -2 (Nrf-2), heme oxygenase (HO-1) expression, by enzyme-linked immunosorbent assay (ELISA) method for the detection of oxidized low density lipoprotein in serum (ox-LDL), endothelin -1 (ET-1), thromboxane B2 (TXB2), 6- keto prostaglandin F1 alpha (6-keto-PGF1 alpha) expression, to illustrate the mechanism of alcohol extract of Schisandra chinensis on treating atherosclerosis. The rat model of atherosclerosis was successfully established, the rats in the model group showed obvious aortic plaque, intimal thickening of the vessel wall, ranging from the number of visible foam cells the plaque. Compared with the model group, the aortic intimal Schisandra alcohol extract group, middle and outer membranes, a relatively complete structure, plaque formation is not obvious, intimal thickening, smooth muscle fibers and mild lesions can be reduced significantly The AS model of rat aortic plaque area percentage of vascular wall area (P0.01 or P0.001), but also can significantly reduce the number of aortic plaque in foam cells (P0.05 or P0.01). Liver pathological observation showed that there were lots of vacuoles in the size of the liver in the model group liver cells, the high dose group and the model control group in the cytoplasmic vacuoles and no obvious lesions. Simvastatin group and ethanol extract of Schisandra chinensis in each dose group compared with the control group, could significantly reduce the content of serum TG (P0.001), significantly decreased the content of LDL-C (P0.001 or P0.01 or P0.05), and high dose group content increased significantly HDL-C (P0.01 or P0.05); simvastatin group and ethanol extracts of Schisandra chinensis group can significantly decrease the serum ALT of rats, the activity of AST (P0.001 or P0.01 or P0.05). The alcohol extract of Schisandra chinensis group can significantly enhance the serum The activity of GSH-PX significantly decreased the content of MDA in serum (P0.001), low dose group can significantly enhance the activity of CAT (P0.001), low dose group can significantly enhance the activity of SOD (P0.01); the mechanism study further showed that the expression of alcohol extract of Schisandra chinensis in each group significant upregulation of rat aortic Nrf-2 protein expression (P0.001), high dose group could significantly upregulate HO-1 protein (P0.01). The alcohol extract of Schisandra chinensis significantly reduced the expression of ox-LDL (P0.01), significantly up-regulated the expression of 6-keto-PGF1 alpha (P0.001), low dose group can significantly reduce the expression of ET-1 (P0.01 or P0.05), and the expression of each dose group of TXB2 had no significant effect. Conclusion Schisandra alcohol extract has obvious anti atherogenic effects can promote lipid metabolism in rats, and its mechanism may be related to the activation of the Nrf-2/ARE pathway, inhibition of ox-LDL, ET-1. White expression regulates the balance of TXB2 and 6-keto-PGF1 alpha in plasma, regulates the level of oxidative stress and maintains the function of endothelial cells.

【学位授予单位】：西南交通大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R285.5

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