| Hyperlipidemia refers to lipid metabolism disorder in human body, causing atherosclerosis with elevating total cholesterol (TC), low density lipoprotein-cholesterol (LDL-C), triglyceride (TG), apolipoprotein B (apo-B) and(or) lipoprotein (a) [(Lp(a)], and lowering high density lipoprotein-cholesterol (HDL-C) and (or) apolipoprotein A (apo-A). Hyperlipidemia is a risk factor in the formation and development of atherosclerosis, and atherosclerosis is the basic pathogenesis of cardiovascular and cerebrovascular disease. A vast volume of research data show that hyperlipidemia is an independent and vital risk factor leading to stroke, coronary heart disease, sudden cardiac death, therefore a vital pathway in controlling hyperlipidemia is essential in preventing cardiovascular and cerebrovascular disease.Acupuncture is a natural therapy, it plays a significant role in the traditional Chinese medicine treatment. It lasts for thousands of years because of its simple, convenient, effective and low-cost operation. Also it gradually becomes outstanding in treating a number of modern diseases as researches prove acupuncture has good result in regulating lipid metabolism in human body. Now, there is more clinical research development in treating hyperlipidemia by means of acupuncture, but less experimental research has been done. By experimental study, the effect of acupuncture in treating hyperlipidemia can be studied in depth with a view to further reveal its working mechanism. It can provide important evidence to support its application in prevention of certain clinical syndrome. This experimental research on the intervention effect of acupuncture on relative factors in hyperlipidemic rat model can provide systematic evidence in preventing hyperlipidemic syndrome. It can provide scientific evidence for treating major diseases by means of acupuncture which is a specialty in Chinese medicine treatment, also it sets a reference in methodology for basic research in Chinese medicine in prevention of major diseases.This research was carried on subsequent to the foundation built by previous researches in the past. Hyerlipidemic rat model was developed by feeding rats with high fat diet. The working mechanism of the effect of acupuncture in rat model in treating hyperlipidemia is studied in depth using modern testing technique.This paper is divided into 3 parts:literature review, experimental research and conclusion.1. Literature reviewA vast volume of literature review relating to hyperlipidermia were read, areas covered including latest research development in the concept of hyperlipidermia, its relationship with related diseases, its pathogenesis and prevention. Analysis on the current conditions of research development in treating hyperlipidermia with Chinese and western medicine has been done, particularly summarizing different ways of treating hyperlipidermia within the country and abroad, and proposal has been made for future research. Acupuncture is believed as one of the effective method in improving hyperlipidermia, therefore more effort should be made to study the working mechanism in depth.2. Experimental study2.1 Method2.1.1 Experimental Groups30 male SD rats [weight (200±20)g], fed by ordinary animal feeds for 1 week, randomly divided into 3 groups according to weight,10 rats a group, viz. blank control group, hyperlipidermic model group, acupuncture group. The rats in blank control group were fed by ordinary animal feeds, all rats in other 2 groups were fed by high fat diet. As from the 6th week, tests were carried out in every other week by drawing blood from the orbital venous sinus of each rat in the blank control group and the model group for checking out TG, TC, HDL-C, LDL-C, calculating the artheriosclerosis index for purpose of comparison, and determining the successful building of hyperlipidermic model. In the 8th week, the hyperlipidermic model was successfully developed. In the following 3 weeks, the rats in the blank control group continued to be fed by ordinary animal feeds, meanwhile, rats in model group and acupuncture group continued to be fed on high fat diet, and acupuncture treatment started once daily on the rats in acupuncture group.2.1.2 Selection of acupuncture points and acupuncture methodPoint selection:"3 lipid points" (Neiguan of Pericardium Meridian of Hand-Jueyin, Zusanli of Stomach Meridian of Foot-Yangming, Sanyinjiao of Spleen Meridian of Foot-Taiyin), Fenglong and Ganshu. Their location was determined by reference to the rats commonly used acupuncture points:Neiguanlocated on the medial aspect of the rat's anterior limb,3mm from wrist joint between ulna and radius bone. Zusanli-located on the rat's hind limb, lateral to the knee joint and 5mm inferior to the fibula small head. Sanyinjiaolocated on the medial aspect of the rat's hind limb,10mm superior to the tip of the internal malleolus. Fenglong-located on rat's lower limb, mid-point of the lower limb and at the posterior border of fibula. Ganshuon the back of the rat,5mm lateral to the spinous process of the ninth thoracic vertebrae.Needling method:For Neiguan, Zusanli, Sanyinjiao, Fenglong, use No.30 one-inch filiform needle (Hwato Brand, produced by the Suzhou Medical Appliance Factory) by oblique insertion for 8mm, the direction of these needle points were inserted towards the centre of the body. For Ganshu, insert needle perpendicularly for 5mm. After needles insertion, turned on the power of the electronic acupuncture treatment instrument and connected pulse current to the Neiguan, Zusanli, Sanyinjiao, Fenglong by clipping output wires to each needle handle of the point group concerned (2 points make up a group) respectively. The electronic acupuncture treatment instrument Model SDZ-II, Hwato Brand, produced by Suzhou Medical Appliance Factory, was used. In application, sparse and dense wave at frequency 5-25 Hz was selected. Proper intensity of stimulation was marked by a slightly muscular twitch around the point. Acupuncture treatment was done continuously for 21 days at 30min per day.2.1.3 Sample handling and testing methodAfter treatment, each group of rats were on fasting for 12 hours overnight as stipulated, for the purpose of testing TG, TC, HDL-C, LDL-C, and calculating atherosclerosis index for future comparison. At the end when all intervention treatments were completed, rats were sedated with sodium pentobarbital (35mg/kg), and then blood was drawn from peritonovein. Blood lipid related indicators (TG, TC, HDL-C, LDL-C), hemorheology related indicators, ET, NO, SOD, MDA, GSH-Px were determined. Rats were executed by dislocated method, a section of liver tissues was removed to test for SOD, MDA, GSH-Px content, and the remaining left hepatic lobule was preserved in 10% formalin solution. They were dehydrated, embedded in paraffin, cut into sections of 3mm thick and stained with hematoxylin and eosin (HE), and then examined under optical microscope for morphological structure.2.2 Result2.2.1 The effect of acupuncture on the weight and general conditions in hyper lipidemic rat model.During the process of experiment, rats in the blank control group were seen to have weight increased steadily in good physical state; they were responsive, able to move freely, and their hair grown with luster. As from the observation obtained from the 5th week, the rats in model group and acupuncture group, when compared with the blank control group, were seen to appear tame, sleepy and with less movement, their hair became fluffy and lost luster, less feces were discharged, their hair turned grey and softer, they looked fatter and weight increased faster.For rats in the model group, their weight increase could still be seen as from the 8th week but growth rate slowed down, they appeared apathetic, less responsive and acted slowly. For rats in the acupuncture group, they were given acupuncture treatment as from the 9th week, and were still fed with high fat diet; their weight increase could still be seen but the rate of increase less than that in the previous weeks. They appeared apathetic, less responsive and acted slowly. After acupuncture treatment for 3 weeks, conditions of rats in acupuncture model had turned better and the rate of weight gain reduced dramatically.2.2.2 The effect of acupuncture on atherosclerosis index (Al) and lipid metabolism in hyper lip idemic rat model.After feeding with high fat diet for 8 weeks, the rats in the model group and acupuncture group (before acupuncture treatment) were found to have lipid metabolism disorder. The results of rats in the model group and acupuncture group (before acupuncture treatment), when compared them with the blank control group, showed a statistically significant effect with a higher TG, TC, LDL-C in serum (P<0.05-0.01), a lower HDL-C in serum (P<0.01), and a higher AI index (P<0.01). By this result, rats in the model group and acupuncture group (before acupuncture treatment) had been confirmed to have lipid metabolism disorder and atherosclerosis after feeding with high fat diet for 8 weeks.After acupuncture treatment for a further 3 weeks, the serum of rats in model group and acupuncture group were tested. The results of the rats in model group, when compared them with the blank control group, showed a statistically significant effect with a higher TG, TC, LDL-C level (P<0.05～0.01), a lower HDL-C (P<0.01), and a higher AI index (P<0.01). The results of the rats in acupuncture group, when compared them with the model group, showed a statistically significant effect with a lower TG, TC, LDL-C level (P<0.05～0.01), a higher HDL-C (P<0.05), and a lower AI index (P<0.01).2.2.3 The effect of acupuncture on liver morphology in hyper lipidemic rat model.Blank control group:the structure of hepatic lobule was clear, central vein was centrally located, border of hepatic plate was smooth, and the hepatic cords were orderly arranged and spread outwards in radius arrangement. No hyperplasia was seen in sinusoidal endothelium cells, fat-storing cells and kupffer cells. Hepacytoplasmic was red stained and hepatocyte nuclear located in central. The structure of the portal tract was seen clear, and bile duct was not damaged. No hyperplasia was seen in fibrous connective tissue. No fatty degeneration and liver inflammation phenomenon could be seen (see Figure 1, Appendix 3). Model group:Diffused hepatocellular fatty degeneration with primarily small bubble fats was visible. There was cell infiltration inside the liver lobules but could not see hepatic fibrosis. Steatotic liver cells were extremely swollen in round shape; mass size was dramatically larger than normal. Cytoplasm was filled with great amount of fat vacuoles (see Figure 2, Appendix 3). Acupuncture group:Fatty degeneration in the liver cells was visible, primarily of small bubble fat. There was slight cell infiltration inside the liver lobule (see Figure 3, Appendix 3).2.2.4 The effect of acupuncture on hemorheology in hyper lipidemic rat model.The blood of hyperlipidemia rats in model group was of high viscosity and hypercoagulability i.e. the blood viscosity increased and hemagglutination built up. After acupuncture treatment for 3 weeks, the blood of the model group and acupuncture group were tested. The result of the blood in model group, when compared with the blank control group, showed a statistically significant effect with a higher High shear rate, Low shear rate, plasma viscosity, hematocrit, and red blood cell electrophoresis time, and red blood cell aggregation index (P<0.05 or P<0.01). The result of the blood in acupuncture group, when compared with model group, showed a statistically significant effect with a lower High shear rate, hematocrit, red blood cell electrophoresis time, and red blood cell aggregation index (P<0.05 or P<0.01). 2.2.5 The effect of acupuncture on oxygen radical in hyper lipidemic rat model.When lipid protein of rats in model group were found elevated, peroxidation also increased, more MDA were generated, but SOD and GSH-Px were found reduced. After acupuncture treatment for 3 weeks, the oxygen radical in the rats of model group and acupuncture group were tested. The result of the model group, when compared with the blank control group, showed a statistically significant effect with a lower SOD and GSH-Px in serum (P<0.05 or P<0.01), and a higher MDA level in serum (P<0.01). The result of the acupuncture group, when compared with the model group, showed a statistically significant effect with a higher GSH-Px level in serum (P<0.05), and a lower MDA level in serum (P<0.01). 2.2.6 The effect of acupuncture on Nitric Oxide (NO) in serum and endothelin (ET) in blood plasma in hyperlipidemic rat model.Hyperlipidemia could cause damage to endothelium leading to endothelial dysfunction, or damage to the function and structure of vascular wall affecting the vascular function. ET and NO played a significant role in the vasoconstriction and vasorelaxation. After acupuncture treatment for 3 weeks, the NO and ET in rats of all groups were tested. The result of NO and ET in the model group, when compared with the blank control group, showed a statistically significant effect with a lower NO in serum and a higher ET in blood plasma (P<0.05 or P<0.01). The result of NO and ET in acupuncture group, when compared with the model group, showed a statistically significant effect with a higher NO in serum and a lower ET in blood plasma (P<0.05).3. ConclusionFeeding high fat diet to the rats in model group could successfully duplicate hyperlipidemic rats. The rats in model group could show all basic characteristics that the hyperlipidermic rats possessed:high lipid in blood, fatty liver, atherosclerosis, concentric obesity etc. This research study confirmed the hemoendothelial function of rats in model group had adversely been affected significantly. These findings had paved an easy way to pursue on. The effect of acupuncture on lipid metabolism disorder in hyperlipidemic rat model group showed a significant improvement. Acupuncture could reduce lipid level in hyperlipidemic rats and atherosclerosis index, enhanced the condition of fatty liver. The effect of acupuncture in rats of hyperlipidemic model showed a significant difference when compared with that of the model group. Acupuncture could show significant effect on hyperlipidemic rats, its mechanism demonstrated that it could lower lipid in blood, improve the atherosclerosis index, adjusting the imbalance of oxidization-anti-oxidant system in the body, and improve hemorrheology, oxygen radicals and condition of fatty liver.
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