| Objective: The purpose of this study was to investigate the target of salidroside in the prevention and treatment of atherosclerosis in Apo E-/-mice and to provide a strong basis for the development of Chinese medicine.Methods: Six weeks old male SPF Apo E-/-(N=30)mice were fed for one week and Western high-fat diet was given 8 weeks to replicate the atherosclerosis model.After successful model establishment,they were randomly divided into model group,positive group,high-dose salidroside group and low-dose salidroside group.Each group has 7 rats;The normal group and model group were given normal saline of equal volume and the positive group was given intragastric administration of atorvastatin(10mg/kg).The high and low dose group of salidroside were given intragastric administration with 100mg/kg and 50mg/kg,respectively.At 8 weeks,During the administration of Apo E-/-mice,all groups were fed with high-fat diet throughout the day.Seven C57BL/6/J mice with the same genetic background were given normal diet as normal group.Observe the change of body weight of each group of mice during the period of administration;after the end of the experiment,the levels of serum lipids(TC,TG,HDL-C,and LDL-C)were detected by enzyme method;HE stain and oil red O staining were used to observe the plaque condition;all macrophages were labeled with F4/80 by immunohistochemistry,M1 macrophages were labeled with i NOS,and M2 macrophages were marked with CD206.The giant plaques were observed according to the expression.The distribution and proportion of phagocytes were detected by ELISA.The levels of M1-type cytokines MCP-1,IL-12p70,IL-6,L-1?and M2-type cytokines IL-10 and IL-13 were detected by ELISA.1.Blood lipid test results: compared with the normal group,the model group TG,TC,HDL-C,LDL-C significantly increased(P <0.01);positive group,high and low doses of salidroside TC,HDL-C,LDL-C Significantly increased(P<0.01);compared with the model group,TG,TC,and LDL-C in the high-dose group of salidroside were significantly lower(P<0.01),and HDL-C was increased(P<0.05);positive group Low dose TG of salidroside was decreased(P<0.05);Compared with the positive group,TC,LDL-C of high-dose salidroside group was significantly lower(P<0.01),and HDL-C was increased(P<0.05).LDL-C decreased in the low-dose group of salidroside(P<0.05).2.Pathological results:(1)HE staining: In the normal group,smooth and intact endothelium was seen,smooth muscle fibers were arranged neatly,cell morphology was intact,no atheromatous plaques were formed in the blood vessels,and no foam cells were seen;atherosclerotic plaque formation and smooth muscle of the media were observed in the model group.The increase of fiber shows a lot of foam cells and cholesterol crystals.In the positive group,the inner membrane was not smooth,no plaque was formed,and the middle membrane became thinner,but the outer membrane elastic plates had different degrees of damage;the salidroside high and low dose groups can be seen in the plaque.A small amount of foam cells had obvious cholesterol crystals,and the smooth muscle tissue layers of the media were clear.Compared with the model group,the degree of fibromuscular proliferation and foam cells in the smooth muscle fibers of the media were all reduced.(2)Oil red O staining: Compared with the normal group,more plaques were observed in the high and low dose groups of the model group,the positive group,and salidroside(P<0.01).Compared with the model group,the high dose of the positive group and salidroside was observed.The plaque area was reduced in all groups(P<0.05).There was no difference in plaque area between the low-dose salidroside group and the model group and the high-dose salidroside group(P>0.05);3.Immunohistochemical results: F4/80 was mainly used for labeling macrophages.Under light microscope,compared with the normal group,the model group,positive group,high dose group of salidroside,and low dose of salidroside were visible Results:brown.The brown coloration showed positive expression of F4/80,i NOS,and CD206 with statistical significance(P<0.01).Compared with the model group,the expression of F4/80 in the positive group and high dose of salidroside was significantly reduced.The expression of CD206 was significantly increased(P<0.01)and the expression of i NOS was decreased(P<0.05).There was no difference between the positive group and the high-dose group of salidroside(P>0.05).Compared with the positive group,Rhodiola rosea The expression of CD206 in the low-dose group of salidroside was higher in the glycoside low-dose group than in the high-dose salidroside group.The expression of CD206 was increased and the expression of CD206 was decreased in the low-dose glucosides group(P<0.01).The amount was significantly reduced with statistical significance(P<0.01).4.ELISA detection of mouse serum M1/M2 cytokine expression levels: Compared with the normal group,the expression of MCP-1,IL-12p70,IL-6 and IL-1? in the model group was significantly increased(P<0.01),IL-10 expression was significantly decreased(P<0.01),The expression of IL-1? was significantly increased in the positive group,high dose group of salidroside,and low dose of salidroside(P<0.01);The expression of IL-10 in the positive group and high dose group of salidroside was significantly increased(P<0.01),The high-dose salidroside group and low-dose salidroside group significantly increased the expression of IL-13(P<0.01).The expression of IL-12p70 was increased in high-dose salidroside group,and the expression of MCP-1 was increased in low-dose salidroside group.Significance(P<0.05);Compared with the model group,the levels of IL-6and IL-1? in the positive group and the high and low dose group of salidroside were significantly reduced,and IL-10 expression was significantly increased.The positive group and Rhodiola rosea The expression of IL-12p70 in the high-dose glycoside group was significantly reduced,IL-13 expression in the high-dose salidroside group was significantly increased,and IL-13 expression in the low-dose salidroside group was significantly reduced,with statistical significance(P<0.01),the positive group and high-dose salidroside group can reduce the level of MCP-1(P<0.05);compared with the positive group,the high and low dose groupof salidroside can be Decrease IL-10 level(P<0.01),salidroside high dose group can increase IL-13 expression(P<0.01),salidroside low dose group can increase IL-12p70 expression,reduce IL-13 expression(P<0.05).Conclusions: Salidroside may have the effect of lowering body weight in atherosclerotic model mice,lowering serum TC,TG,and LDL-C,increasing HDL-C,thereby reducing foam cells,and reducing plaque formation.Its mechanism of action may be related to the regulation of lipid metabolism,adjustment of macrophage M1/M2 polarization,suppression of inflammatory response,stabilization of plaques and development of anti-atherosclerosis. |
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