电针对动脉粥样硬化模型兔血清MIF、IL-10影响的实验研究

发布时间：2018-06-25 04:44

本文选题：动脉粥样硬化 + 电针　；参考：《辽宁中医药大学》2017年硕士论文

【摘要】：目的:观察电针内关、关元、足三里穴对动脉粥样硬化模型兔的干预作用,通过检测各组实验兔血清中MIF、IL-10表达水平,探讨电针内关、关元、足三里穴抑制动脉粥样硬化炎症反应的作用机制。材料与方法:普通级雄性新西兰兔26只,体重2.0-2.5kg,3月龄。采用高脂饲料喂养联合颈总动脉球囊损伤术法建立动脉粥样硬化模型。将26只实验兔随机分成两组:空白对照组(7只)、造模组(19只)。空白对照组,给予普通饲料进行喂养;造模组,予高脂饲料喂养,并于造模第4周施以颈总动脉球囊损伤术,术后继续高脂饲料喂养4周。进行造模干预8周后,从两组各随机选取1只实验兔处死,取其颈总动脉采用HE染色法观察动脉粥样斑块形成情况,以确认模型复制成功与否。确认造模成功后,空白对照组剩余实验兔(6只)继续如前喂养;采用随机数字表法将造模组剩余实验兔(18只)均分为三组,分别为:AS模型组(6只)、电针干预组(6只)、阳性药物对照组(6只),均予高脂饲料喂养,并开始治疗,6日为1疗程,各疗程间休息1日,连续进行4个疗程。治疗方法:电针干预组予针刺内关穴、关元穴、足三里穴治疗,进针后,于同侧“内关穴、足三里穴”,关元穴和兔尾非经非穴的针柄处接通电针治疗仪,予疏密波,强度以实验兔肢体轻颤、能够耐受不烦躁为度,电流1mA,留针20 min,1次/d;阳性药物对照组予阿托伐他汀钙片1mg/kg/d制成悬浊液灌胃治疗。第4疗程结束后,于实验兔耳缘静脉采血待测,再行耳缘静脉空气栓塞法处死后,予颈动脉取材,采用HE染色法制备病理切片,观察颈动脉组织的病理形态学变化,用酶联免疫吸附法(ELISA,Enzyme Linked Immunosorbent Assay)测定兔血清中巨噬细胞移动抑制因子(MIF)和白介素-10(IL-10)的含量,探讨电针刺激内关、关元、足三里穴对动脉粥样硬化的治疗作用机制。结果:1.光镜下观察:空白对照组兔颈总动脉内膜完整连续,无炎症细胞浸润及泡沫细胞形成;AS模型组兔颈总动脉内膜明显增厚,内皮细胞不连续,单核细胞浸润及聚集增多,大量泡沫细胞聚集成斑块;电针干预组及阳性药物对照组兔颈总动脉内膜增厚,内皮细胞不连续,可见单核细胞浸润,有泡沫细胞形成,较AS模型组病变程度明显减轻。2.与AS模型组相比较,电针及阳性药物治疗均可下调AS兔血清中MIF的含量。AS模型组与各组结果比较有显著差异(P0.05);比较电针干预组与阳性药物对照组血清中MIF水平,无明显差异(P0.05)。3.与AS模型组相比较,电针及阳性药物干预均可上调AS兔血清中IL-10的含量。AS模型组与各组比较有显著差异(P0.05);比较电针干预组与阳性药物对照组血清中IL-10水平,无明显差异(P0.05)。结论:1.电针内关、关元、足三里穴,可通络活血化痰,从而抑制动脉粥样斑块形成。2.电针内关、关元、足三里穴,抑制兔血清中促炎因子MIF的表达,促进血清中抗炎因子IL-10的表达,可能通过通络活血化痰抑制炎症反应,进而控制动脉粥样硬化的发展。
[Abstract]:Objective: To observe the effect of electroacupuncture Neiguan, Guan yuan and Zusanli on the atherosclerotic rabbit model, and to explore the mechanism of the effect of electroacupuncture Neiguan, Guan yuan and Zusanli on the inhibition of atherosclerotic inflammation by detecting the MIF and IL-10 expression levels in the serum of all experimental rabbits. Materials and methods: 26 normal male New Zealand rabbits, weighing 2.0-2 .5kg, 3 month old. A model of atherosclerosis was established by feeding high fat diet combined with balloon injury of common carotid artery. 26 rabbits were randomly divided into two groups: blank control group (7) and model group (19). The blank control group was fed with ordinary feed, the model group, high fat feed were given, and the common carotid artery was given for fourth weeks. After the operation, the high fat diet was continued for 4 weeks. After 8 weeks of model intervention, 1 rabbits were randomly selected from two groups, and the carotid artery plaque formation was observed by HE staining, to confirm the success of the Model replica. After the model was successful, the remaining rabbits in the blank control group (6) continued to be fed as pre feeding. The experiment rabbits (18 rats) were divided into three groups by random digital table method: AS model group (6), electroacupuncture intervention group (6), positive drug control group (6), all were fed with high fat diet, and the treatment was given, the 6 day was 1 courses, and 1 days in each course. The treatment method: acupuncture intervention group was given acupuncture. Guan acupoint, Guan yuan acupoint, Zusanli treatment, after entering the needle, at the same side "Neiguan point, Zusanli point", Guan yuan point and the tail of the rabbit at the needle handle of non acupoints through the acupuncture treatment instrument, the intensity of the density wave, the experimental rabbit limb tremble, can tolerate no irritability, current 1mA, 20 min, 1 /d; positive drug control group to Atorvastatin Calcium Tablets 1mg/ After the fourth course of the fourth course of treatment, after the end of the fourth course of treatment, the blood collection of the rabbit ear vein was measured. After the ear vein air embolism was executed, the carotid artery was selected and the pathological section was prepared by HE staining method. The pathological changes of the carotid tissue were observed, and the enzyme linked immunosorbent assay (ELISA, Enzyme Linked Immunosorbent Assay) was used. The contents of macrophage migration inhibitory factor (MIF) and interleukin -10 (IL-10) in rabbit serum were measured, and the therapeutic mechanism of electroacupuncture stimulation of Neiguan, Guan yuan and Zusanli on atherosclerosis was discussed. Results: 1. light microscopy showed that there was no inflammatory cell infiltration and foam cell formation in the blank control group. No inflammatory cell infiltration and foam cell formation; AS model. The intima of the common carotid artery in the rabbits was thickened, the endothelial cells were discontinuous, the infiltration and aggregation of monocytes increased, and a large number of foam cells were aggregated. The intima of the common carotid artery in the electroacupuncture intervention group and the positive drug control group were thickened, the endothelial cells were discontinuous, the monocyte infiltration and the formation of foamy cells were visible, and the degree of pathological changes in the AS model group was significantly reduced. Comparison between light.2. and AS model group, electroacupuncture and positive drug treatment could reduce the content of MIF in AS rabbit serum.AS model group compared with the results of each group was significantly different (P0.05); compared with the positive drug control group and the positive drug control group, there was no significant difference in serum MIF level (P0.05).3. and AS model group, electroacupuncture and positive drug intervention could be compared. There was significant difference between the.AS model group and the IL-10 in the serum of AS rabbit serum (P0.05), and there was no significant difference in the serum IL-10 level between the electroacupuncture intervention group and the positive drug control group (P0.05). Conclusion: 1. electroacupuncture Neiguan, Guan yuan, Zusanli point can collaterals and activate blood phlegm, thus inhibiting the formation of atherosclerotic plaque of the.2. electroacupuncture Neiguan, Guan yuan, foot. The three li point inhibits the expression of pro-inflammatory factor MIF in rabbit serum and promotes the expression of anti inflammatory factors IL-10 in serum. It may inhibit the inflammatory reaction through activating blood and activating blood and eliminating phlegm, and then controlling the development of 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;R-332

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