| Objectives: The purpose of this study is tried to explore the mechanism of improvement of aortic sinus plaque for Apo E-/- mice aortic sinus atherosclerosis by exlectroacupuncture thearpy. The study was executed electroacupuncture intervention in Apo E-/- mice, and then the expression of IL-6 protein in the aortic sinus plaque and liver morphology changes were observed and analyzed to achieve objectives.Methods: The 6-week-old 38 Apo E-/- mice were randomly divided into two groups, 34 mice in high fat and high-cholesterol diet group(HF&HC diet group) and 4 in moderate-fat diet group(MF diet group). The ingredients of HF&HC diet contain 21% fat and 1.25% cholesterol and MF diet contained 10% fat diet. After feeding HF&HC diet for 14 weeks, 4 mice in the HF&HC diet group were randomly selected as the HF group, HC group and MF group, and then they were immediately killed to collect plasma, take aortic sinus tissues, examine the conventional hematoxylin-eosin(HE) staining, frozen HE staining and oil red O staining. At the same time, comparison of feeding different diet on mice, the results will be detected by Lee’s index, cholesterol, the hepatosomatic index(HI), body fat ratio(BFR), and pathological changes in hepatic and cardio arteries. The remaining 30 Apo E-/- mice of HF&HC diet group were randomly divided into 10 of each group: 10 in the drug control group(F group), 10 in the electroacupuncture treatment group(EA group), 10 in the model control group(M group). All groups continually were given HF&HC diet. Meanwhile, five C57 BL / 6J mice were fed normal diet as the control group(BC group). M group were given oral treatment with Simvastatin(辛伐他汀), 40mg/kg.d, EA group received acupuncture therapy by Neiguan, Zusanli, Sanyinjiao, and Shenmen. After 11 weeks experimental study, the morphological changes of aortic sinus plaques in mice were examinded by HE staining, immunohistochemical staining for each group to understand the expression of IL-6.Results:(1) The main findings was revealed that both M group and F group were displayed mainly a large number of porridge-like necrotic core plaque and a lot of cholesterol crystals. plaque fibrous cap in M group is very thin, elastic fibers disappeared. Plaque ruptured phenomenon occured in F group, the content of elastic fibers in F group were significantly less than in EA group. After acupuncture treatment, the fibrous cap became more thicker than before, and also some were not yet formed, They displayed stable plaques. The area of fatty Plaques in EA group were smaller than in both M group and F group. The plaques mainly contained large amounts of foam cells in EA group, thus this phenomenon explained that plaques were still in gradually changing stage. A large porridge-like necrotic core and a lot of cholesterol crystal were displayed apoptosis of foam cells in EA group so that it was indicated the blood vessels stayed in atherosclerotic plaques stage. In all above phenomena, we may conclude the acupuncture therapy could protect vascular endothelium, retrain the development of plaque, and treat more effective than drug do.(2) Aortic sinus IL-6 immunohistochemical staining : The aortic sinus plaques IL-6 was mainly distributed in the fibrous cap. IL-6 was secreted and expressed by inflammatory cells such as monocytes, macrophages, smooth muscle cells. One of study results was found that a large amount of IL-6 was distributed around medial smooth muscle cells in F group. In addition, some findings were medial smooth muscle cell migration and exhausted with less expression tan particles in HF&HC diet group within F group. In M group a large number of IL-6 was located in the aortic fibrous cap and aortic valve. In EA group IL-6 was distributed in the fibrous cap and a blood vessel membrane; there were a smaller amount of IL-6 expression in the fibrous cap with significantly less than in M group; and the expression of IL-6 on the aortic valve had significantly been reduce. In M group plaque apparently contained a wide range of nuclear and cholesterol crystals necrosis; accompanied with massive necrosis and apoptosis. In F group plaque contained more cholesterol crystals and necrotic core than others while EA group was obviously no necrotic core plaques and cholesterol crystals. In summary, the studies may confirm that electroacupuncture therapy could protect the intima, inhibit smooth muscle cell proliferation and migration, prevent the development of plaque and make more effective treatment than drug could do.Conclusions Electroacupuncture therapy could regulate liver, intravascular lipid, energy metabolism, anti-oxidation, anti-inflammatory, inhibit the migration of vascular smooth muscle cell proliferation, prevent vascular plaque rupture, protect hepatic cells and smooth muscle cells, check out foam cells apoptosis and necrosis, and so on. Following above advantages, the conclusions were that the electroacupuncture therapy would be the way to prevent the development of atherosclerotic plaque as playing a role to gain health spleen and benefits qi and to have effectiveness on of dissipating phlegm and removing blood stasis. |
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