电针对动脉粥样硬化家兔肝脏中胆固醇7α-羟化酶表达的影响
发布时间：2018-10-14 09:00
【摘要】：目的:通过观察电针对动脉粥样硬化家兔肝脏中胆固醇7α-羟化酶蛋白及基因表达的影响,探讨针刺治疗动脉粥样硬化新的作用机制,为临床针刺治疗动脉粥样硬化及相关疾病提供新的实验依据。材料与方法:将26只雄性新西兰兔,体重在2.5kg±0.5kg左右,月龄2~3个月,分笼饲养,适应性喂养1周,随机选取7只为空白组,余下19只为造模组。空白组喂以普通饲料,造模组给予高脂饲料,喂养4周后,造模组家兔行颈总动脉球囊损伤术,术后继续隔日高脂饲料喂养4周,从空白组和造模组各随机选一只家兔做颈动脉病理切片,采用HE染色法观察颈动脉粥样斑块病理形态,确定造模成功与否。造模成功后,将造模组18只兔子随机分为3组:模型组、电针组和西药组,每组6只。模型组不给予任何治疗;西药组将含有阿托伐他汀钙片的混悬液灌胃给药;电针组选内关、关元和足三里穴进行电针治疗,每日1次,每次留针20min,6日为1个疗程,疗程间休息1日,共4个疗程。治疗结束以后,在家兔耳缘静脉采血,检测各组家兔血脂水平;取颈动脉做病理切片,采用HE染色法观察颈动脉粥样斑块病理形态;取家兔肝脏组织,采用Western blot法检测家兔肝脏中CYP7A1蛋白表达水平;采用RT-PCR检测家兔肝脏中CYP7A1m RNA表达水平。结果:1.血脂水平:与空白组相比,模型组家兔血液中CHO、TG以及LDL含量均升高,HDL含量下降,有统计学意义(p0.01);与模型组比较,电针组和西药组家兔血液中CHO、TG以及LDL含量均显著降低,HDL含量升高,有统计学意义(p0.01);电针组与西药组相比,无显著差异(P0.05)。2.CYP7A1蛋白表达:与空白组相比,模型组家兔CYP7A1蛋白表达水平下降,具有统计学意义(p0.01);与模型组比较,电针组和西药组家兔CYP7A1蛋白表达水平显著升高,具有统计学意义(p0.01);电针组与西药组相比,无显著差异(P0.05)。3.CYP7A1m RNA表达:与空白组相比,模型组家兔CYP7A1m RNA的表达水平下降,具有统计学意义(p0.01);与模型组相比,电针组、西药组家兔CYP7A1m RNA的表达水平显著升高,具有统计学意义(p0.01);电针组与西药组相比,无显著差异(P0.05)。结论:1.电针内关、关元、足三里穴能改善动脉粥样硬化家兔血脂水平。2.电针内关、关元、足三里穴能促进家兔肝脏中CYP7A1m RNA和蛋白的表达,这可能是电针治疗动脉粥样硬化的机制之一。
[Abstract]:Objective: to observe the effect of electroacupuncture on cholesterol 7 伪 -hydroxylase protein and gene expression in the liver of atherosclerotic rabbits, and to explore the new mechanism of acupuncture in treating atherosclerosis. To provide a new experimental basis for clinical acupuncture treatment of atherosclerosis and related diseases. Materials and methods: 26 male New Zealand rabbits, weighing about 2.5kg 卤0.5kg, were kept in cage for 2 ~ 3 months and fed adaptively for one week. Seven rabbits were randomly selected as blank group and the remaining 19 as model group. The blank group was fed with common feed and the model group was given high fat diet. After 4 weeks of feeding, the rabbits in the model group were treated with carotid artery balloon injury, and were fed with high fat diet every other day for 4 weeks. A rabbit was selected randomly from blank group and model group to make pathological sections of carotid artery. The pathological morphology of carotid atherosclerotic plaque was observed by HE staining to determine whether the model was successful or not. After successful modeling, 18 rabbits in model group were randomly divided into 3 groups: model group, electroacupuncture group and western medicine group with 6 rabbits in each group. The model group was not given any treatment; the western medicine group was given intragastric administration of suspension containing Atto vastatin calcium tablets; the electroacupuncture group was treated with electroacupuncture at Neiguan, Guanyuan and Zusanli points, once a day for 20 mins each time, and 6 days as a course of treatment. Rest 1 day between the courses, a total of 4 courses. After the treatment, blood was collected from the auricular vein of rabbits at home, blood lipid levels were detected in each group, carotid artery was taken as pathological section, pathological morphology of carotid atherosclerotic plaque was observed by HE staining, liver tissue was taken from rabbits. The expression of CYP7A1 protein in rabbit liver and CYP7A1m RNA in rabbit liver were detected by Western blot and RT-PCR respectively. The result is 1: 1. Blood lipid level: compared with the blank group, the blood CHO,TG and LDL contents in the model group were increased, the HDL content was decreased (p0.01), and compared with the model group, the blood lipid level in the model group was significantly higher than that in the model group. The contents of CHO,TG and LDL in the blood of rabbits in electroacupuncture group and western medicine group were significantly decreased, and the content of HDL was increased (p0.01), but there was no significant difference between electroacupuncture group and western medicine group (P0.05). The expression of 2.CYP7A1 protein: compared with blank group, the expression of 2.CYP7A1 protein in electroacupuncture group was significantly lower than that in western medicine group (P0.05). Compared with the model group, the expression of CYP7A1 protein in the electroacupuncture group and western medicine group was significantly increased (p0.01), the expression of CYP7A1 protein in the electroacupuncture group was significantly higher than that in the western medicine group (p0.01), the expression of CYP7A1 protein in the electroacupuncture group was significantly higher than that in the western medicine group (p0.01). No significant difference (P0.05). 3.CYP7A1m RNA expression: compared with the blank group, the expression of CYP7A1m RNA in the model group decreased significantly (p0.01); compared with the model group, the expression of CYP7A1m RNA in the electroacupuncture group and western medicine group increased significantly. There was no significant difference between electroacupuncture group and western medicine group (P0.05). Conclusion: 1. Electroacupuncture at Neiguan, Guanyuan and Zusanli can improve blood lipid levels in atherosclerotic rabbits. Electroacupuncture at Neiguan, Guanyuan and Zusanli can promote the expression of CYP7A1m RNA and protein in rabbit liver, which may be one of the mechanisms of electroacupuncture in treating atherosclerosis.
【学位授予单位】：辽宁中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R245
本文编号：2269997
[Abstract]:Objective: to observe the effect of electroacupuncture on cholesterol 7 伪 -hydroxylase protein and gene expression in the liver of atherosclerotic rabbits, and to explore the new mechanism of acupuncture in treating atherosclerosis. To provide a new experimental basis for clinical acupuncture treatment of atherosclerosis and related diseases. Materials and methods: 26 male New Zealand rabbits, weighing about 2.5kg 卤0.5kg, were kept in cage for 2 ~ 3 months and fed adaptively for one week. Seven rabbits were randomly selected as blank group and the remaining 19 as model group. The blank group was fed with common feed and the model group was given high fat diet. After 4 weeks of feeding, the rabbits in the model group were treated with carotid artery balloon injury, and were fed with high fat diet every other day for 4 weeks. A rabbit was selected randomly from blank group and model group to make pathological sections of carotid artery. The pathological morphology of carotid atherosclerotic plaque was observed by HE staining to determine whether the model was successful or not. After successful modeling, 18 rabbits in model group were randomly divided into 3 groups: model group, electroacupuncture group and western medicine group with 6 rabbits in each group. The model group was not given any treatment; the western medicine group was given intragastric administration of suspension containing Atto vastatin calcium tablets; the electroacupuncture group was treated with electroacupuncture at Neiguan, Guanyuan and Zusanli points, once a day for 20 mins each time, and 6 days as a course of treatment. Rest 1 day between the courses, a total of 4 courses. After the treatment, blood was collected from the auricular vein of rabbits at home, blood lipid levels were detected in each group, carotid artery was taken as pathological section, pathological morphology of carotid atherosclerotic plaque was observed by HE staining, liver tissue was taken from rabbits. The expression of CYP7A1 protein in rabbit liver and CYP7A1m RNA in rabbit liver were detected by Western blot and RT-PCR respectively. The result is 1: 1. Blood lipid level: compared with the blank group, the blood CHO,TG and LDL contents in the model group were increased, the HDL content was decreased (p0.01), and compared with the model group, the blood lipid level in the model group was significantly higher than that in the model group. The contents of CHO,TG and LDL in the blood of rabbits in electroacupuncture group and western medicine group were significantly decreased, and the content of HDL was increased (p0.01), but there was no significant difference between electroacupuncture group and western medicine group (P0.05). The expression of 2.CYP7A1 protein: compared with blank group, the expression of 2.CYP7A1 protein in electroacupuncture group was significantly lower than that in western medicine group (P0.05). Compared with the model group, the expression of CYP7A1 protein in the electroacupuncture group and western medicine group was significantly increased (p0.01), the expression of CYP7A1 protein in the electroacupuncture group was significantly higher than that in the western medicine group (p0.01), the expression of CYP7A1 protein in the electroacupuncture group was significantly higher than that in the western medicine group (p0.01). No significant difference (P0.05). 3.CYP7A1m RNA expression: compared with the blank group, the expression of CYP7A1m RNA in the model group decreased significantly (p0.01); compared with the model group, the expression of CYP7A1m RNA in the electroacupuncture group and western medicine group increased significantly. There was no significant difference between electroacupuncture group and western medicine group (P0.05). Conclusion: 1. Electroacupuncture at Neiguan, Guanyuan and Zusanli can improve blood lipid levels in atherosclerotic rabbits. Electroacupuncture at Neiguan, Guanyuan and Zusanli can promote the expression of CYP7A1m RNA and protein in rabbit liver, which may be one of the mechanisms of electroacupuncture in treating atherosclerosis.
【学位授予单位】：辽宁中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R245
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