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To study the effects of CLLC on experimental atherosclerosis quails and rats models as well as the cell models in vitro., in which the protection effects of CLLC-containing serum to the injured human umbilical vein endothelial cells induced by H2O2 was observed. In addition, the mechanism of CLLC on treating atherosclerosis was also discussed.Methods:in vivo, experiments:(1) the hyperlipidemia quails model and atherosclerosis quails model were established by high fat diet fed method (cholesterol 1.5, ordinary diet fed 78.5, peanut oil 4, lard 16). The quails were randomly devided into five groups as follows:model group (saline), positive group (lovastatin 7.48 mg/kg), CLLC high dose group (40.00 g/kg), CLLC medium dose group (26.67 g/kg), CLLC low dose group (13.33 g/kg), respectively. The cupsules were given by oral administration once a day. Meanwhile, the five groups were given with high fat diet while the control group was given with ordinary diet. The quails received a 12-weeks administration course and the concentrations of totle cholesterol (TC), triglycreride (TG), high density lipoprotein-cholesterol (HDL-C), low density lipoprotein-cholesterol (LDL-C), oxidation low density lipoprotein (oxLDL) in serum were measured in the 4 th week and the 12th week, respectively. Naked-eye examination and histological observation with light microscopy on aorta,,left truncus brachiocephalicus artery as well as right truncus brachiocephalicus artery was performed. After that, the liver index was calculated and the concentrations of NO, NOS, Ang II, cGMP, TXB2, ET,6-keto-PGF1, MMP-1, MMP-9, TIMP-1, EMMPRIN in serum were measured. Furthurmore, the circulating endothelial cells (CEC) in serum was counted and the concentrations of CD40, CD40L, TGF-β, sVCAM, sICAM in serum were also analyzed. (2) Twenty male SD rats and 20 female SD rats were given high diet (4. cholesterol,0.5. sodium tauroglycocholate,0.2. propylthiouracil,10 lard,85.3 ordinary diet), then rat AS model was established by injuring right common carotid artery and intramuscular injection of vitamin D3. The rats were randomly devided into five groups as follows:model group (saline), positive group (lovastatin 7.48 mg/kg), CLLC high dose group (40.00 g/kg), CLLC medium dose group (26.67 g/kg), low dose group (13.33 g/kg), respectively, while control group (8 rats) was given ordinary diet. The cupsules were given by oral administration once a day, after that, specimens of blood and right common carotid artery were collected, indications including whole blood viscosity, plasma viscosity, blood-lipid, t-PA, PAI, IL-6, IL-8 and TNF-were examined along with histopathologic studies and observation of the change of artery walls with microscopy. (3) Male rats were chosen and randomly devided into five groups as follows:control group (saline), positive group (troxerutin 48 mg/kg), CLLC high dose group (40.00 g/kg), CLLC medium dose group (26.67 g/kg), CLLC low dose group (13.33 g/kg), respectively. Mixed thrombus model was established by injuring common carotid artery with electric stimulus and the effect of capsules on thrombosis was studied. KM mouse were chosen and randomly devided into five groups as follows:control group (saline), positive group (troxerutin 48 mg/kg), CLLC high dose group (40.00 g/kg), CLLC medium dose group (26.67 g/kg), CLLC low dose group (13.33 g/kg), respectively.For studying the effect of capsules on thrombosis, acute lung embolism model was established by mixed thrombus. In vitro. method:(1) the cupsules were given by oral administration once a day (capsule 40.00 g/kg), the rats received a 7-day administration course and their serum were collected at 1.5 h after administration on the 7 th day. (2) Injured human umbilical vein endothelial cell model induced by H2O2 was established to observe the protection effects of CLLC in different dose with MTT method. The concentrations of SOD, MDA, NO, MMP-2, MMP-9, EMMPRIN, TGF-β, CD40 and CD40L in cell supernatant were measured.Results:In vivo.experiments:(1) the CLLC could dramatically decrease the concentration of liquid(lipid?) in serum and decrease the rate of AS plaque. after the the intervension treatment of the capsule drug, the concentrations of Ang II, TXB2, ET, MMP-1, MMP-9, TIMP-1, EMMPRIN in serum were dereased while the concentrations of NO, NOS, cGMP,6-keto-PGF1 were increased. Compared with the model group, the capsule could dramatically decrease the amount of CEC in serum, decrease the concentrations of CD40, CD40L, TGF-B, sVCAM, sICAM (P<0.05 or P<0.01). (2) after the the intervension treatment of the capsule, the whole blood viscosity of rats as well as the concentrations of TG, TC, LDL-C, PAI, IL-6 were decreased while the concentrations of HDL-C, IL-10 and TNF-were increased. The formation of arterial wall plaque was reduced; the deposits of foam cells and lipid were shorten; the amount of macrophage in AS plaque was decreased; the time of the formation of carotid artery inner membrane thrombus induced by electric injury was dramatically extended; the rat acute lung thrombus were inhibited; the duration of asthma induced by ADP mixed thrombosis were shortened (P<0.05 or P<0.01). In vitro:CLLC could dramatically prompt the cytoactive of human umbilical vein endothelial cells injured by H2O2, decrease the concentrations of MDA, MMP-2, EMMPRIN, TGF-β, CD40 and CD40L and increase the concentrations of SOD in cell supernatant.Conclusion:CLLC could dramatically decrease the concentrations of lipid in serum and reduce risk of the formation of AS plaque. The mechanism could be associated with the inflammation inhibition, protecting the function of vascular endothelial cells and supressing the formation of AS plaque.
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