Study On Inflammatory Factors Of Apo E^-/-mice With Jing-hu-tong-mai Prescription Based On Nur77

Purpose:Observe the effect of Jing-hu-tong-mai Prescription to blood fat and atherosclerosis plaque of Apo E^-/-mice.Study on the effect of Jing-hu-tong-mai Prescription to Nur77 expression and inflammatory Factor secretion in Atherosclerosis mice and its effect on macrophages.To explore the possible mechanism that Jing-hu-tong-mai Prescription interven in ATH and to provide experimental basis for the prevention and treatment of ATH.Method:32 SPF Apo E^-/-mice were fed with high-fat Western diet for 8 weeks?including fat 21%,cholesterol 0.15%?to establish atherosclerosis model in Apo E^-/-mice.The models were randomly divided into model group,positive group,high dose group of Jing-hu-tong-mai Prescription,middle dose group of Jing-hu-tong-mai Prescription and low dose group of Jing-hu-tong-mai Prescription,6 rats in each group.6C57BL/6/J mice served as normal control group,and were fed with common feed.The normal group and the model group were given 0.3ml/ / day of saline,the positive group was given atorvastatin for 10mg/kg/ days,and the high,middle and low dose groups were given /kg/ days for 18.20 g,/kg/ days of 9.10 g and /kg/ days of 4.55 G,1times a day for 8 weeks.Serum cholesterol?TC?,triglyceride?TG?,low density lipoprotein cholesterol?LDL-C?and high-density lipoprotein cholesterol?HDL-C?levels in mice were detected.HE staining was used to observe the vascular structure and morphology of the aorta and calculate the plaque area.Masson staining was used to observe and evaluate the collagen fibers in the plaques.ELISA was used to detect serum tumor necrosis factor-??TNF-??,interleukin-1?IL-1 beta?and interleukin-6?IL-6?in mice.Toll-like receptor 4?Toll-like receptors 4,TLR4?,adenosine triphosphate binding cassette transporter A1?ATP binding cassette transporter A1,ABCA1?,Nur77 expression in the aorta were detected by immunohistochemistry.Result:?1?Four blood test:compared with normal group,elevated serum lipid level in the model group;compared with the model group,the Jing-hu-tong-mai Prescription high,medium and low dose groups and positive drug group lipid levels decreased,HDL-C levels increased,with significant difference?P<0.01?.?2?The results of HE staining:normal group intima integrity,without atherosclerotic plaque;vascular model group showed a large number of plaques,cell disorder,vascular wall thickness is not uniform;compared with the model group,positive group and Jing-hu-tong-mai Prescription high and low dose group of aortic plaque area decreased,and has statistically significant difference?P<0.01?.Positive drug group and the Jing-hu-tong-mai Prescription Prescription high,medium and low dose group between the patch area had no significant difference;Jing-hu-tong-mai Prescription high dose group compared with Hutong vein patch area landscape of low dose group and small plaque area,with significant difference?P<0.01?.?3?Masson staining results:there was no plaque in the normal group and no collagen fibrils in the plaque.Compared with the model group,positive drug group and increased Jing-hu-tong-mai Prescription high,medium and low dose group of mice aorta plaque collagen fiber content was statistically significant?P<0.01?.Compared with the Jing-hu-tong-mai Prescription high dose group,low dose group,the collagen content in plaque positive drug group and the Jing-hu-tong-mai Prescription was lower,with statistical difference?P<0.05?.?4?ELISA and Luminex to detect the expression of TNF-alpha,IL-1 beta,IL-6?IL-17A:model group IL-1 beta.TNF-alpha and IL-6 level is significantly higher than normal group,with statistical difference?P<0.01?;compared with the model group,positive group and Jing-hu-tong-mai Prescription IL-1 beta level high,medium and low dose group were decreased,with statistical difference?P<0.01?,positive group and Jing-hu-tong-mai Prescription high,medium and low pulse dose group IL-1 betalevels were not significantly different.There was no significant difference in serum IL-17 A level in each group.?5?Immunohistochemical method was used to detect the expression of TLR4,ABCA1 and Nur77 in aorta:TLR4 staining showed that the aorta in normal group no obvious TLR4 expression,expression of the model group and the Jing-hu-tong-mai Prescription of high,medium and low dose group were in a large number of TLR4.Compared with the model group,positive drug group and the Jing-hu-tong-mai Prescription high,medium and low dose group TLR4 expression decreased,there was significant difference?P<0.01?;Jing-hu-tong-mai Prescription dose TLR4 expression than the Jing-hu-tong-mai Prescription low dose group,there was significant difference.ABCA1 staining showed that there was no significant ABCA1 expression in aorta of normal group.There was a large number of ABCA1 expression in model group and drug intervention group.The positive group and the king Hutong pulse high,medium and low dose group the expression of ABCA1 with the model group,there was no statistically significant difference?P>0.05?.Nur77 staining showed no obvious Nur77 expression in the aorta of normal group,and Nur77 expression was found in model group and drug intervention group.Compared with the model group,positive drug group and the Jing-hu-tong-mai Prescription high,medium and low dose group Nur77 expression was increased,there was significant difference?P<0.01?;Jing-hu-tong-mai Prescription high dose Nur77 expression than the king Hutong vein of low dose group,and there was statistical difference?P<0.05?.Conclusion:The anti atherosclerotic effect of Jin-hu-tong-mai Prescription may be to regulate the secretion of inflammatory factors and regulate blood lipid by interfering with Nur77 intervention in TLR4 signaling pathway,thus inhibiting the occurrence and development of inflammation and interfering with the development of atherosclerotic plaque,and ultimately to protect the blood vessels and resist atherosclerosis.