Experimental Study On Chinese Caterpillar Fungus Oral Solution Improving Atherosclerosis In Apolipoprotein E Knockout Mice

OBJECTIVE:To investigate the effect of Chinese Caterpillar Fungus oral solution on reducing atherosclerotic plaque in apolipoprotein E knockout mice and its possible mechanism by prophylactic administration.METHODS:ApoE^-/-mice of 40 male C57BL/6J backgrounds were randomly divided into model group?normal saline?,atorvastatin positive drug group?10mg/kg?,and Chinese Caterpillar Fungus oral solution low dose group?0.4g/kg?,medium dose group?1.2 g/kg?,high dose group?3.6 g/kg?.Another 8 male C57BL/6J mice were used as blank control?normal saline?.After 8 weeks of high-fat diet feeding,serum of mice was taken to measure TC,TG,LDL-C,HDL-C,MDA,SOD,IFN-?,TNF-?,IL-5,IL-12p70 content by using biochemical method and enzyme-linked immunosorbent assay.The blood vessels of the mouse aortic arch were isolated.The size of the plaque in the lumen was compared with a microscope;the area of the chest and abdominal aorta was compared with the area of the oil red O staining;the frozen area of the aortic sinus was compared with the oil red O staining to compare lipid deposition;Paraffin sections of arterial sinuses were compared with H&E,Masson,and Sirius red staining to compare changes in the pathology of the arterial endometrial tissue and collagen fibers;paraffin sections of the aortic sinus were quantitatively analyzed by immunohistochemical staining for macrophage infiltration and galectin-3 and vascular smooth muscle actin expression.RESULTS:1.Compared with the blank group,ApoE^-/-mice can obviously develop atherosclerotic plaque in the aorta after 8 weeks of high-fat diet.2.After drug intervention,the levels of TC,TG,LDL-C,MDA,IFN-?,TNF-?,IL-5,IL-12p70were decreased,and the levels of SOD activity and HDL-C were increased in the Chinese Caterpillar Fungus oral solution group and positive drug group.The difference between the model group and the Chinese Caterpillar Fungus oral solution administration group was statistically significant?P<0.05?.3.The results of oil red O staining in the thoracoabdominal aorta of mice showed that after 8 weeks of high-fat diet,the mice in the model group had significantly more plaques than the control group.Compared with the model group,the positive drug group and Cordyceps sinensis oral solution were low.The ratio of aortic plaque area to total vascular area decreased by 31.4%?P<0.01?,18.8%?P<0.05?,and 20.0%?P<0.05?,respectively.The difference was statistically significant.The results of oil red O staining at the aortic sinus showed that after 8 weeks of high-fat diet,lipid deposition in the model group was significantly more than that of the blank control group;compared with the model group,the positive drug group and Cordyceps sinensis oral solution were given low and medium doses.The ratio of plaque area to total lumen area of the aortic sinus in the drug group decreased by 33.7%?P<0.01?,23.4%?P<0.05?,and 33.3%?P<0.05?,respectively.The difference was statistically significant..Aortic sinus H&E staining results showed that compared with the control group,the aortic lumen of the model group narrowed,the intima thickened and protruded into the lumen,and a large number of vascular smooth muscle cells proliferated under the intima,showing a lipid plaque.Blocks and foam cells aggregated;compared with the model group,the ratio of the thickened intima to the luminal area of the aortic sinus in the mice in the low and middle dosage groups of the positive drug group and the Cordyceps sinensis oral solution decreased by 33.8%?P<0.01?,25.4%?P<0.05?and 29.5%?P<0.01?,the difference was statistically significant.Masson and Sirius red staining of the aortic sinus showed that compared with the blank group,the expression level of collagen fibrils in the aortic sinus of mice in the model group increased significantly;compared with the model group,the positive drug group and Cordyceps sinensis oral solution The contents of aortic medial collagen fibers in the low and middle dose groups decreased by 27.5%?P<0.01?,21.5%?P<0.05?,24.9%?P<0.05?,and 28.3%?P<0.01?,respectively.,16.3%?P<0.05?,21.3%?P<0.05?,the difference was statistically significant.The results of immunohistochemistry on paraffin sections of the aortic sinus showed that compared with the blank group,the infiltration of macrophages in the aortic sinuses,the expression of galectin-3,and the intima-smooth muscle actin in mice of the model group were significantly increased.The level increased;compared with the model group,the infiltration of macrophages in the aortic sinus of the mice in the positive drug group and the Cordyceps sinensis oral solution decreased by 33.8%?P<0.01?and 19.9%?P<?,respectively.0.05),27.7%?P<0.01?.The expression level of galectin-3 decreased by 30.2%?P<0.01?,19.3%?P<0.05?,21.9%?P<0.05?.-The expression of smooth muscle actin decreased by 37.5%?P<0.01?,20.3%?P<0.01?,and 28.3%?P<0.01?.The difference was statistically significant.CONCLUSION:1.ApoE^-/-mice atherosclerosis model was successfully induced by high-fat diet feeding in this experimental method.2.Chinese Caterpillar Fungus oral solution can reduce the serum levels of TC,TG,LDL-C,MDA,IFN-?,TNF-?,IL-5,IL-12p70 and increase the levels of SOD and HDL in ApoE^-/-mice.3.Chinese Caterpillar Fungus oral solution can reduce the plaque size in the aorta and aortic sinus of ApoE^-/-mice,reduce the infiltration of macrophages in the aorta and the expression of galectin-3,and inhibit blood vessels smooth muscle cells migrate and increase the content of aortic collagen fibers.4.Chinese Caterpillar Fungus oral solution can improve atherosclerosis in ApoE-/-mice.The mechanism may be to reduce lipid and inflammatory cytokines in the serum,thereby reducing lipid deposition and macrophage infiltration in the aorta,in the end reduce plaque area and improve AS.