Correlation between FcγRⅢA and Aortic Atherosclerotic Plaque Destabilization in ApoE Knockout Mice and Intervention Effects of Effective Components of Chuanxiong Rhizome and Red Peony Root

Objective： To explore the correlation between Fc γ R IIIA （CD16A） and aortic atherosclerotic plaque destabilization in apoE knockout （apoE KO） mice and the intervention effects of effective components of Chuanxiong Rhizome and Red Peony Root. Methods： Eight 8-week-old male C57BL/6J mice were selected as the control group. Forty 8-week-old male apoE KO mice were randomly divided into the model group, apoE KO ＋ intraperitoneal injection immunoglobulin group （IVIG）, apoE KO ＋ simvastatin group （Sm）, apoE KO ＋ high dosage of Xiongshao Capsule （XSC,芎芍胶囊） group （XSCH）, and apoE KO ＋ low dosage of XSC group （XSCL）, 8 mice in each group. Mice in the control group were put on a normal diet, and others were fed with a high-fat diet. After 10-week different interventions, monocyte CD16 expression was detected by flow cytometry, aortic matrix metalloproteinase-9 （MMP-9） mRNA expression was detected using reverse transcription- polymerase chain reaction, and serum tumor necrosis factor （TNF）-a level was detected using enzyme-linked immunosorbent assay. Results： Compared with the control group, monocyte CD16 expression, aortic MMP-9 mRNA expression, and serum TNF-a level in the model group increased obviously （P〈0.01）. Injections of apoE KO mice with intraperitoneal immunoglobulin during a 5-day period significantly reduced the monocyte CD16 expression, aortic MMP-9 mRNA expression, and serum TNF-a level （P〈0.01 or 0.05） over a 10-week period of high-fat diet. Indices above in the Sm group, XSCH group, and XSCL group decreased in a different degree. Of them, the aortic MMP-9 mRNA expression in XSCH group was lower than that in Sm group （P〈0.05） and the monocyte CD16 expression and serum TNF-a level showed no significant difference between XSCH group and Sm group （P〉0.05）. Correlation analyses suggested positive correlation between monocyte CD16 expression and aortic MMP-9 mRNA expression or serum TNF-a level in IVIG group, XSCH group, and XSCL group. Conclusions： Fc γ/R IliA mediates systemic inflammation in the progression of coronary heart disease with blood stasis syndrome. XSC could stabilize atherosclerotic plaque by suppressing inflammation and its target was relative with Fc γ RIIIA.

Expression and Location of Intracellular Tissue Factor in Atherosclerosis Stable Plaque of ApoE^(-/-) Mice

In the ApoE^-/- mouse model of atherosclerosis （AS） stable plaque, the expression and location of intracellular tissue factor （TF） in the cellular components of AS stable plaque were investigated in order to explore the cellular mechanism of AS thrombosis. Pathological changes of the stable plaque were observed under a microscope. The expression of TF protein was examined in aortic stable plaque of mice by using immunohistochemistry. Color image planimetric system was used to analyze the histological components of the stable plaque and the TF distribution. Under the confocal microscope, the intracellular TF location in the stable plaque of mice was observed. The results showed the cellular area was the major part of stable plaque （67.36%±6.52%, P〈0.01）. The percentage of total area occupied by cellular area was significantly larger than atheromatous gruel and acellular area （P〈0.01）. Macrophages and smooth muscle cells （SMC） were major cells in the cellular area. The percentage of total area occupied by SMC was significantly larger than by macrophages （P〈0.01）. Multiple linear regression analysis showed there was a positive correlation between TF area and SMC area （r=0.616, P=-0.008）, and no correlation was found between TF area and macrophage area （r=0.437, P=0.08）. Pictures of color image planimetric analysis of TF and SMC were merged to highlight areas with co-localization （yellow）, it was concluded that the process could be a cell-mediated TF expression in the stable plaque. SMC may be the major source of TF in AS without plaque rupture.

Establishment of a rat model with diet-induced coronary atherosclerosis

Coronary atherosclerotic disease is a serious disease in humans,but no suitable animal model is available currently for further studies.We used apolipoprotein E gene knockout（ApoE KO） rats to induce hypercholesterolemia through a special high cholesterol/bile salt diet（Paigen diet）,then analyzed aortic and coronaiy atherosclerosis lesions and the myocardial injury in order to establish a novel small animal model of coronary atherosclerosis.Plasma cholesterol of ApoE KO rats increased 7.6-fold compared with wild-type rats after 8 weeks on the Paigen diet.After 10 to 12 weeks of subsisting on the Paigen diet,ApoE KO rats developed mild aortic atherosclerosis with severe coronary atherosclerosis.Hematoxilyn and eosin staining showed that 11 out of 12 ApoE KO male rats had right coronary artery atherosclerosis,7 of them were〉70%occluded.Oil Red O（Lipid Stain）,Mac2 immuno-staining and Masson＇s tnchrome staining demonstrated substantial amounts of lipid,macrophages and collagen fibers in coronary atherosclerosis plaques.In addition,ApoE KO male rats had severe myocardial focal lesions with cholesterol ester as the main component in the lesions.In conclusion,ApoE KO rats developed severe hypercholesterolemia,coronary atherosclerosis and myocardial cholesterol ester deposition after subsisting on the Paigen diet and can be used as a novel animal model for studies on cholesterol metabolism and coronary atherosclerotic disease.