| Objective:To explore the effect of Tongsaimai on the atherosclerotic plaque of carotid artery, adiponectin and plasma lipid on the basis of successful establishment of the model of carotid atherosclerotic plaque in rats, and to judge curative effect of the medicine and to provide a reliable experiment- al basis for the clinical treat.Methods:1. Establishment of animal model and grouping: totally 90 sprague dawley rats (sanitary degree), after a week adaptive feed, 10 rats were selected randomly to sacrifice, then the rest were divided into 4 groups randomly: normal group, model group, prevention group, therapy group (each group n=20). model group, protected group, therapy group were fed on high-fat food, and were subjected to the intramuscular injection of the vitamin D3 needle (70*105 IU/kg/d) continuously for four days at the beginning of the experiment; normal group were given common forage. The prevention group started to administrate by gastric infusion (the 6.22g pharmacognosy/kg) on the day that they were given fat-rich-diet, while control group, model group, therapy group, injected with the same volume of normal saline every day for 8 weeks. Since the 9th week, the therapy group was gaster-poured by medicine (the 6.22g pharmacognosy/kg), the other groups were given the same treatment as that previously until 12 weeks. All animal freedom drinking and were separated to rise.2. High-fat food: 3% cholesterols, 0.5% sodium cholate, 0.2% propyl–thiouracil, 5% refined sugars, 10% lard, 81.3% common forage.3. Sample collection: At the beginning of formal experiment, 10 rats were selected randomly to sacrifice. Then at the 8th weekends and the 12th weekends, 10 rats were selected randomly to sacrifice from every group. The organization of carotid arteries and blood serum were kept for microscopic pathology and index examination at each time point.4. Detection method of adiponectin: Enzyme linked immunosorbent Assay was adopted to detect adiponectin, data was measured strictly according to the introduction and was read in an hour. Used standard concentration as Y-axis, OD as X-axis, the standard curve was established by software package of Curve Expert 1.3. Then we can look up the concentration of adiponectin in the standard curve according to OD value of the serum sample.5. Detection method of the blood-fat: Enzyme linked immunosorbent assay was used to measure the serum total cholesterol (TC), triglyceride (TC) and low density lipoprotein (LDL).6. Statistical treatment: The data obtained were represented as mean±SD, and analyzed by software package of SPSS 13.0 software after managed through the database of EXCEL. Significance was tested with partnership designed t-test and analysis of variance according to the comparison of two or three groups. There was statistically significance P<0.05.RESULTS:1. Compared with the initial value, the level of Adiponectin in four groups were lower in varying degrees and showed statistics difference (P<0.05) at the different time points. Compared with the initial value, the level of TC,TG,LDL in model groups were higher at different degree and showed statistics difference (P<0.05) at the different time.2. The changes of the different factors in the prevention group:At the 8th weekend, the level of adiponectin in model group, prevention group were all significantly lower than that of the normal group and showed statistics difference (P<0.05). The level of adiponectin were significantly different among the model group and the prevention group,the normal group (P<0.05). At the 12th weekend, the level of adiponectin in model group, prevention group were all lower than that of the normal group and showed statistics difference (P<0.05). The level of adiponectin in prevention group were significantly higher than that of the model group and showed statistics difference (P<0.05). At the 8th weekend, the TC, TG, LDL concentrations in model group, prevention group were all higher than that of the normal group and showed statistics difference (P<0.05). The level of them in prevention group were lower than that of the model group and showed statistics difference (P<0.05). At the 12th weekend, the level of them in model group, prevention group were all higher than that of the normal group and showed statistics difference. The prevention group was lower than that of the model group.3. The changes of the different factors in the therapy group:At the 8th weekend, the level of adiponectin in model group, therapy group was all lower than that of the normal group and showed statistics difference (P<0.05). There was no difference between therapy group and model group (P>0.05). At the 12th weekend, the level of adiponectin in model group, therapy group were all lower than that of the normal group. The level of Adiponectin in the therapy group was higher than that of the model group and showed statistics difference (P<0.05).At the 8th weekend, the TC, TG, LDL concentrations in model group, therapy group were all higher than that of the normal group and showed statistics difference (P<0.05). There was no different between in therapy group and model group (P>0.05). At the 12th weekend, the TC, TG, LDL concentrations in model group, therapy group were all higher than that of the normal group. The level of them in the therapy group was lower than that of the model group and showed statistics difference (P<0.05).4. The morphological observation of Carotid artery in four groups observed under microscope: (HE staining)At the 8th weekend, in model group and therapy group, there were considerable xanthoma cell, fibroblast and collagen aggregated under carotid artery undarterium. Lipid-necrosis-cores were obvious which showed blanc or pallideflavens lipid-plaque and yellow lipochondria. In prevention group, undarterium was not obviously integrity, thicken or adipose infiltration. There was little change in normal group changed. At the 12th weekend, there was obvious atherosclerotic plaque of carotid artery in model group. The amount of unsteady atherosclerotic plaque and lack of impartiality plaque was increased. Some lipid-cores had already detached or disappeared. The prevention group treatment of Tongsemai, undarterium was not obviously integrity, thicken or adipose infiltration. In therapy group, the degree of thicken significantly was reduced, the mount of xanthoma cell or Surroundings burning cell was decreased, and the area of lipid-plaque was reduced, but still has a few fibroblast and collagen. It showed that atherosclerotic plaque was smaller, its degree alleviated and its stability increased.Conclusion:1. We took an advanced method that induced by high fat diet combine with the vitamin D3 intraperitoneal injection, and established a successful model of the carotid atherosclerotic elastic arteries in rats.2. The prevention group and therapy group treatment of tongsaimai can effectively increase the level of adiponectin and decrease the level of blood lipid. It can decrease the progression during the formation of atherosclerotic plaque and enhance the stability of atherosclerotic plaque of rats.3. Tongsaimai had significant curative effect in the prevention group and the therapy group. It had preventive effect in prevention group is superior to that of therapy group. So it's a kind of ideal Chinese patent medicine to cure or prevent atherosclerotic plaque, and is worthy of generalization and application in clinic. |
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