敌百虫加速大鼠动脉粥样硬化模型的建立及染料木素干预

发布时间：2018-06-01 07:36

本文选题：染料木素 + 动脉粥样硬化　；参考：《南华大学》2011年硕士论文

【摘要】：目的:利用高脂饮食和低剂量、低毒性有机磷农药敌百虫腹腔注射建立SD大鼠动脉粥样硬化模型,观察染料木素抗大鼠动脉粥样硬化作用,并检测血脂、丙二醛含量等生化指标,初步探讨其作用机制,为染料木素的合理用药提供实验和理论依据。方法: 健康SD大鼠56只分为七组,其中普通饲料喂养(普食)即溶媒对照组(1 ml·kg~(-1)·day~(-1))、高剂量染料木素(genistein ,Gen)对照组(5mg·kg~(-1)·day~(-1))和阳性药物辛伐他汀对照组(5mg·kg~(-1)·day~(-1))三组,高胆固醇高脂饲料喂养(简称高脂)加敌百虫即模型对照组[10%的二甲亚砜(DMSO),1ml·kg~(-1)·day~(-1)]、低剂量Gen对照组(0.5mg·kg~(-1)·day~(-1))、高剂量Gen对照组(5mg·kg~(-1)·day~(-1))和阳性药辛伐他汀对照组(5 mg·kg~(-1)·day~(-1))四组。 2周为大鼠称一次体重,根据体重调整给药剂量。给药后继续按原饲料方案喂养8周。在实验给药前和给药后第4、8周分别断尾采血,使用怡成全血葡萄糖测定仪检测大鼠血糖(Glu),用德国罗氏诊断有限公司试剂盒检测大鼠血清总胆固醇(TC)、甘油三酯(TG)、高密度脂蛋胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)、C反应蛋白(CRP)含量;实验第4、8周末用分光光度法检测血清对氧磷酶(PON1)活性,用Griess试剂显色法测定大鼠血清总一氧化氮(NO),硫代巴比妥法检测大鼠血清中丙二醛(MDA)浓度;第8周末处死所有动物,采用羟肟酸铁比色法测定大鼠全血乙酰胆碱酯酶(Acetylcholine esterase, AchE)活性,摘取大鼠肝脏及主动脉弓,肉眼和苏木素-伊红(HE)染色后观察各组大鼠主动脉弓及肝脏病理形态学变化;同时摘取心脏分离其周围组织后称重,计算其脏器指数。使用免疫组化SP法检测主动脉血管壁内皮型一氧化氮合酶(eNOS)的蛋白表达情况。结果: 1实验大鼠一般状况普食组大鼠毛色光泽,活动自如,活泼,体重增长较快;高脂喂养加敌百虫的四组大鼠随着实验进程,毛色松散暗淡,活动减少,其中给予低、高剂量Gen和辛伐他汀的三组大鼠精神状态明显好转,体重都呈现先降低在增长,且实验期间体重变化不大。 2血脂含量变化第4、8周末,普食喂养的三组中大鼠血清血脂含量、HDL-C/TC与AI值均无显著差异(P0.05);高脂喂养加敌百虫的四组大鼠血清TC、LDL-C、HDL-C含量与AI值均较普食喂养的三组明显升高(P0.05或P0.01或P0.001),而HDL-C/TC值明显降低(P0.05或P0.01)。各组大鼠血清TG含量无显著差异(P0.05)。Gen能明显降低TC、LDL-C,使AI降低,HDL-C/TC比值升高。 3大鼠GLU含量变化 Gen各剂量组和辛伐他汀均能明显降低动脉粥样硬化大鼠血糖水平(P 0.05或P0.001)。 4血清CRP含量变化高剂量Gen和辛伐他汀能明显降低动脉粥样硬化大鼠血清炎症因子CRP的含量(P0.05)。 5血清MDA含量变化高剂量Gen和辛伐他汀能明显降低血清MDA水平。 6血清NO含量变化高脂喂养加敌百虫的四组大鼠血清NO含量较普食喂养的三组明显升高(P0.05或P0.01);高脂喂养加敌百虫的四组大鼠中,给予Gen和辛伐他汀组与高脂喂养加敌百虫模型对照组大鼠血清NO含量均无显著差异(P0.05)。 7血清PON1含量变化 Gen提高As大鼠血清PON1活性(P0.05或P0.01),存在量效关系。 8组织形态观察肝脏与主动脉弓形态学观察,Gen的各剂量组和辛伐他汀均能在一定程度上对血管壁具有保护作用。 9 Gen对主动脉eNOS蛋白表达的影响免疫组化结果:Gen各剂量组和辛伐他汀均能在一定程度上增加eNOS表达。结论: 1.成功建立敌百虫加速的SD大鼠As模型。 2.Gen能明显抑制敌百虫加速的SD大鼠As的发生,改善其肝与主动脉弓形态变化。其可能的机制与逆转血脂障碍,通过抑制过氧化脂质作用抗氧化,通过降低炎症因子CRP抗炎,并参NO体系、eNOS与PON1酶的调节有关。
[Abstract]:Objective: to establish the atherosclerotic model of SD rats by intraperitoneal injection of high fat diet and low toxic organophosphorus pesticide didiworm intraperitoneally, observe the effect of genistein on atherosclerosis and detect the biochemical indexes of blood lipid and malondialdehyde content, and explore the mechanism of its action to provide experiment and reason for the rational use of genistein. On the basis.
Method:
56 healthy SD rats were divided into seven groups, of which common feed was fed (1 ml. Kg~ (-1). Day~ (-1)), high dose genistein (genistein, Gen) control group (5mg kg~ (-1) day~) and positive drug simvastatin control group (-1). The two methylene sulfoxide (DMSO), 1ml / kg~ (-1) / day~ (-1)) of the model control group, the low dose Gen control group (0.5mg kg~ (-1) day~ (day~)), the high dose control group (kg~) and the positive drug simvastatin control group (1ml) four groups.
The rats were weighed once for 2 weeks, and the dosage was adjusted according to the weight of the rats. After the drug was given for 8 weeks, the blood was collected before and after the 4,8 week, and the blood glucose (Glu) was detected by the Yicheng whole blood glucose analyzer, and the serum total cholesterol (TC) was detected by the German Roche Diagnostics Ltd kit. The content of triglyceride (TG), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C) and C reactive protein (CRP), the activity of serum to oxyphosphatase (PON1) was detected by spectrophotometric method at the end of 4,8 at the end of the experiment, and the serum total nitric oxide (NO) was determined by the chromogenic reagent of Griess reagent, and malondialdehyde (MDA) in the serum of rats was detected by thiobarbituric method. All the animals were killed at the end of the eighth week. The activity of Acetylcholine esterase (AchE) in the whole blood of rats was measured by ferrous hydroxamic acid colorimetry. The liver and aortic arch of rats were extracted, the naked eye and hematoxylin eosin (HE) were stained to observe the pathological changes of the aortic arch and liver in each group, and the circumferential separation of the heart was also taken. The organs were weighed and the Viscera index was calculated. The expression of endothelial nitric oxide synthase (eNOS) in the aortic wall was detected by immunohistochemical SP.
Result:
1 general condition of experimental rats
The four groups of rats in the four groups of rats with high fat feeding and stinworm were given low, high dose and simvastatin. The three groups of rats with high dose and simvastatin showed a significant improvement in the mental state of the rats, and the body weight decreased first and the body weight changed during the experiment. Not much.
Changes of blood lipid content in 2
At the end of the 4,8 weekend, there was no significant difference in serum lipid content in the three groups of the rats fed by universal diet (P0.05). The levels of TC, LDL-C, HDL-C and AI in the serum of four groups of rats fed with high fat feeding and the HDL-C/TC were significantly higher than those of the three groups feeding on the universal diet (P0.05 or P0.01 or P0.001). There was no significant difference in TG content (P0.05)..Gen significantly decreased TC and LDL-C, reduced AI and increased HDL-C/TC ratio.
Changes of GLU content in 3 rats
Gen dose and simvastatin could significantly reduce the blood glucose level of atherosclerosis rats (P 0.05 or P0.001).
Change of serum CRP content in 4
High dose of Gen and simvastatin could significantly reduce the content of serum inflammatory factor CRP in atherosclerotic rats (P0.05).
Change of serum MDA content in 5
High dose of Gen and simvastatin significantly decreased serum MDA level.
Change of serum NO content in 6
The serum NO content of four groups of rats fed with high fat feeding and a total of the three groups were significantly increased (P0.05 or P0.01), and there was no significant difference in the content of NO in the serum of the high fat feeding and the four groups of the simvastatin group and the high fat feeding and the rat model control group (P0.05).
Change of serum PON1 content in 7
Gen increased the activity of serum PON1 (P0.05 or P0.01) in As rats, and there was a dose effect relationship.
8 observation of tissue morphology
Morphologic observation of liver and aortic arch showed that all doses of Gen and simvastatin could protect the vascular wall to some extent.
The effect of 9 Gen on the expression of eNOS protein in the aorta
Immunohistochemical results: Gen dose and simvastatin could increase eNOS expression to some extent.
Conclusion:
1. the As model of SD rats was successfully established.
2.Gen can obviously inhibit the occurrence of As in SD rats accelerated by DTM and improve the morphological changes of the hepatic and aortic arch. Its possible mechanism is related to reversing lipid disorders, inhibiting the antioxidant activity of lipid peroxide, reducing inflammation factor CRP by reducing inflammation and regulating the regulation of eNOS and the regulation of PON1 enzyme in the NO system.
【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R363

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