The Effect Of Red Yeast Rice On Intestinal Inflammation And Barrier Function In Atherosclerotic Mice Model

ObjectiveTo study the protecting effect of red yeast rice on intestinal inflammation and intestinal barrier function in atherosclerotic mice modelMethods1.Animals and Diets:18 8-week-old male ApoE^-/-mice weighed 22±2 g were used for this study.After 1 week adaptive feeding,ApoE^-/-mice were randomly divided into three groups:blank control group,atherosclerosis model group and red yeast group.The red yeast group was intragastrically administered with 0.34 g/kg.d per day.The other groups were administered with normal saline.The atherosclerotic model group and the red yeast group were fed with a high-fat,high-cholesterol diet?containing 15%lard+0.25% cholesterol?.The experimental period was 12 weeks.After the last gavage,the mice were fasted for 12 h,and the eyeballs were taken for blood collection.The blood was centrifuged and serum was taken.The mice were perfused with the heart.After perfusion,aorta,colon and cecum were collected on ice and stored at-80°C.2.Quantification of atherosclerotic plaque:The aorta was stained by Oil red O and the area of atherosclerotic plaque was quantified.3.Determine the effect of red yeast on blood lipids:use automatic biochemical analyzer to determine total cholesterol,tryglyeride,high density lipoprotein cholesterol and low density lipoprotein cholesterol.4.Hematoxylin eosin?HE?was used to observe the pathological change of the colon.5.Ultrastructural change of colonic tissue in mice were observed by transmission electron microscopy.6.Western blot?WB?was used to detect the expression levels of tight junction proteins occludin and JAM-1 in small intestine.7.WB was used to detect the expression level of tumor necrosis factor-??TNF-??and interleukin 1??IL-1??in small intestine.8.16S rDNA high-throughput sequencing for molecular ecology analysis of intestinal flora in cecal contents.Results1.After the oil red O staining of the mouse aorta,analysis and statistics were performed.The plaque area of the blank control group,the atherosclerosis model group and the red yeast group was?5.56±1.45?%,?22.46±8.33%?and?7.00±2.65?%respectively.Compared with the blank control group,the plaque area of the model group significantly increased?P<0.05?.Compared with the atherosclerosis model group,the plaque area in the red yeast group was significantly decreased?P<0.05?.2.Compared with the blank control group,serum cholesterol was significantly increased in the model group?P<0.05?.The cholesterol in the red yeast group was significantly lower than that in the model group?P<0.05?.The red yeast group was higher in density than the blank control group.The protein was significantly decreased?P<0.05?;the low-density lipoprotein was significantly lower in the red yeast group than in the blank control group and the model group?P<0.05?.3.The results of HE staining showed that there was no pathological change in the small intestine mucosa of the blank control group,and the villus and gland were intact.Compared with the blank control group,the intestinal mucosal epithelium was lost in the atherosclerosis model group,the intestinal villi were unclear,the mucosa was congested,and the small intestine was small.The villus height was decreased by 30.44%compared with the control group?P<0.05?.The intestinal villi were mildly lost after the red yeast intervention,the mucosa was basically normal,and the small intestine villus height increased by 44.21%compared with the model group?P<0.05?.4.Electron microscopy showed that the colonic microvilli were abundant and arranged neatly.The columnar epithelial cells were structurally intact and the intercellular junction structure was normal.Compared with the blank control group,the atherosclerosis model group was tightly connected,the desmosome connection was unclear,and the microvilli length was shortened by 53.52%,the difference was significant?P<0.05?;compared with the model group,intercellular junction structure of the red yeast group was normal,and the microvilli-rich microvilli length increased by 47.62%,and the difference was significant?P<0.05?.5.Compared with the blank control group,the expression of occludin and JAM-1 was significantly decreased in the atherosclerosis model group?P<0.05?.Compared with the model group,the protein expression of occludin and JAM-1 in the red yeast group was significantly increased?P<0.05?.6.The expression level of TNF-?and IL-1?in the model group was significantly higher than those in the control group?P<0.05?.The expression level of TNF-?and IL-1? protein in the red yeast group was significantly lower than those in the model group?P<0.05?.7.For the abundance of Firmicutes: model group > control group > red yeast rice group.LEf Se analysis showed that Bacteroides has a significant difference in the red yeast group.Spearman correlation analysis showed that Lachnospiracea_incertae_sedis was negatively correlated with plaque area;Anaeroplasma was negatively correlated with low-density lipoprotein cholesterol;Alistipes was positively correlated with total cholesterol and low-density lipoprotein cholesterol.The abundance of Alistipes was significantly lower in the red yeast group than that of the model group,while the abundance of Anaeroplasma was significantly higher in the red yeast group than in the model groupConclusion1.Red yeast rice can retard the formation of atherosclerotic plaque,which could result from its effect on reduce cholesterol,restoring the balance of intestinal microbiota,protecting intestinal barrier function and reducing intestinal inflammation.2.The significantly altered abundance of Alistipes and Anaeroplasma by red yeast rice indicated their potential function in retarding atherosclerotic plaque formation,which could serve as potential target for atherosclerosis prevention.