| Objective:By making a model of atherosclerosis in diabetic ApoE-/-mice and intervening with allicin,the effect of allicin on this animal model and its mechanism of action were studied,and the mechanism of allicin and glucose and lipid metabolism in protecting the endothelium Relevance of disorders,oxidative stress response,and inflammatory response.Provide experimental support for clinical prevention and treatment of diabetic macroangiopathy.Methods:Eighty eight-week-old ApoE-/-male mice were selected,weighing 25±5g.Randomly divided into a control group(n=20)and an experimental group(n=60).The control group does not do any treatment and is fed with ordinary feed;the experimental group will be injected with STZ,carotid artery cannula,and 3%methionine high fat through the abdominal cavity.Diet,making a mouse animal model of diabetic atherosclerosis,and feeding them adaptively for 1 week after modeling.Divide diabetic mice into model group(double distilled water,0.15ml/d,n=20),allicin low-dose group(allicin,10mg/kg,0.15ml/d,n=20),high-dose allicin Group(Allicin,20mg/kg,0.15ml/d,n=20),to simulate the preventive effect of drugs,after 8 weeks of gavage,blood samples were taken to determine blood lipids(TG,LDL,TC),blood sugar,Hcy Concentration,and then the mice were sacrificed,carotid artery and aortic tissues were taken for HE staining,general oil red 0 staining,oil red 0 staining,Masson staining,Sirius scarlet staining detection,IL-6,MMP-9 expression.Results:1.Blood lipids:1.1 Serum TC:Compared with the control group,the serum TC concentration was significantly increased,with a statistically significant difference(all P<0.01);the serum TC concentration of the allicin high and low dose groups was significantly higher than that of the model group There was a statistically significant difference(all P<0.01);the effect of allicin in high-dose group on reducing serum TC concentration was better than that in low-dose group,and there was a statistical difference between the two groups(P<0.05).1.2 Serum HDL:Compared with each group,the change trend of serum HDL concentration is not obvious,and there is no significant statistical difference(all P>0.05).1.3 Serum LDL:Compared with the control group,the serum LDL concentration was significantly increased,with a statistically significant difference(both P<0.01);the serum LDL of the allicin high and low dose groups was significantly reduced compared with the model group,with a significant Statistical difference(all P<0.01);the effect of allicin high-dose group on reducing serum LDL concentration is better than that of allicin low-dose group,with significant statistical difference(P<0.01).1.4 Serum TG:Compared with the control group,the serum TG concentration of the model group and the low-dose allicin group were significantly increased,and there were significant statistical differences(all P<0.01);the serum TG concentration in the high-dose allicin group was slightly higher than that of the control group There was a decrease,but no significant statistical difference(P>0.05);the serum TG concentration of the allicin low-dose group was lower than that of the model group,but no significant statistical difference(P>0.05);the serum TG concentration ratio of the allicin high-dose group The model group was significantly reduced,with statistically significant differences(P<0.01);the serum TG concentration in the high-dose allicin group was significantly lower than that in the low-dose allicin group,with statistically significant differences(P<0.01).2.Serum Hcy:Compared with the control group,the serum Hcy concentration of the model group was significantly increased,with a statistically significant difference(P<0.01);compared with the control group,the allicin high and low dose groups showed no statistical difference in serum Hcy(all P>0.05);the serum Hcy concentration in the high-and low-dose allicin group was significantly lower than that in the model group,with significant statistical differences(both P<0.01);the serum Hcy in the high-and low-dose allicin group was not statistically significant Difference(P>0.05).3.Blood glucose:The blood glucose of mice in the model group has maintained a high blood glucose state during the experiment(≥16.7mmol/L),which is significantly different from the control group(P<0.01).The mice in the high and low dose groups of allicin After 1 week of intervention,the blood glucose decreased,and the blood glucose was significantly reduced compared with the model group,with significant statistical differences(both P<0.01);the high-dose allicin group was significantly lower in the lower blood glucose group,with significant statistics Difference(P<0.01)4.Body weight:Compared with the control group,the model group and the allicin high and low dose groups showed significant weight loss,and there were significant statistical differences(both P<0.01);the model group mice maintained a downward trend in body weight The mice in the high-and low-dose allicin groups recovered their weight of gavage.5.Histopathological morphology results5.1 HE staining of carotid tissueIn the control group(a),the cells were neat and orderly,the shape was clear,the structure was complete,the lumen was stenosis grade I,the plaque area was<40%(22.16±6.07),the lipid nuclei were small,the fiber cap was thick,and the plaque was stable;Model group(b)Cell arrangement is disordered,morphology is fuzzy,structure is incomplete,lumen stenosis is grade Ⅳ,plaque area>40%(92.81±1.02),large lipid nucleus,thin fiber cap,a large number of foam cells,lipid crystals,plaques Very unstable;the low-dose allicin group(c)is slightly disordered,slightly fuzzy in shape,slightly intact in structure,luminal stenosis grade Ⅱ,plaque area>40%(42.48±0.98),fat The nucleus is too large,the fiber cap is slightly thin,foam cells are visible,and the plaque is slightly unstable;the high-dose allicin group(d)has neatly arranged cells,clear morphology,and complete structure;the lumen is narrow grade II and the plaque area<40%(38±0.44),small fat nucleus,thick fiber cap,stable plaque.5.2 Detection of aortic plaque load in diabetic mice by the method of general oil red 0The control group(a)has a lighter aortic plaque load,while the model group(b)has a severer aortic plaque load.This diabetic mouse model is prone to unstable atherosclerotic plaques.The aortic plaque load was slightly more severe in the low-dose allicin group(c),and the aortic load was significantly reduced in the high-dose allicin group(d).5.3 Results of carotid tissue oil red 0 stainingThe lipid content of the model group(b)was significantly higher than that of the control group(a),with a statistically significant difference(P<0.01);the lipid content of the allicin high(d)and low-dose group(c)Compared with the model group,they were significantly reduced,with significant statistical differences(all P<0.01);the lipid content of the allicin high-dose group was significantly reduced compared with the low-dose group,with a significant statistical difference(P<0.01).5.4 MASSON staining results of carotid tissueCompared with the control group(a),the model group(b)collagen fibers were significantly reduced,with a significant statistical difference(P<0.01);allicin high(d),low dose group(c)compared to the model group collagen fibers Significantly increased,with significant statistical differences(both P<0.01);the collagen fiber in the high-dose allicin group was significantly higher than that in the low-dose group,and there was a significant statistical difference between the two groups(P<0.01).5.5 Carotid tissue Sirius scarlet staining resultsThe control group(a)has a lot of plaque collagen content,mainly type I collagen fibers,and the plaque is stable;the model group(b)plaque is extremely unstable,and its collagen content is significantly reduced,compared with the control group type Ⅰ/Ⅲ Collagen ratio decreased significantly,with significant statistical difference(P<0.01);plaque collagen content in low-dose allicin group(c)was increased compared with model group,and type Ⅰ/Ⅲ collagen ratio increased,with significant statistical difference(P<0.01);plaque in the high-dose allicin group(d)is relatively stable,and its collagen content is increased compared with the model group,mainly type I collagen,and the ratio of type Ⅰ/Ⅲ collagen to the model group and the low-dose allicin group The comparisons were significantly higher,with significant statistical differences(P<0.01,P<0.05).5.6 Immunohistochemical results5.6.1 IL-6 Model group(b)compared with the control group(a)IL-6 expression was significantly increased,with a statistically significant difference(P<0.01);garlicin high(d),low dose(c)Compared with the model group,the expression of IL-6 was significantly reduced,with significant statistical differences(all P<0.01);the expression of IL-6 in the high-dose allicin group was significantly lower than that in the low-dose group,compared between the two groups There are significant statistical differences(P<0.01).5.6.2 MMP-9 Model group(b)compared with the control group(a)MMP-9 expression was significantly increased,with a statistically significant difference(P<0.01);allicin high(d),low dose group(c)Compared with the model group,the expression level of MMP-9 was significantly reduced,with a significant statistical difference(P<0.01);the expression level of MMP-9 in the high-dose allicin group was significantly lower than that in the low-dose group,and the comparison between the two groups was significant Statistical difference(P<0.01).Conclusion:1.8-week-old male ApoE-/-mice injected STZ intraperitoneally can successfully induce diabetes,which is basically similar to the diabetic state of humans.ApoE-/-mice promote the occurrence and development of atherosclerosis and accelerate the formation of unstable plaques in the diabetic state.2.Compared with non-diabetic ApoE-/-mice,the blood lipids(TG,LDL,TC),blood glucose,and Hcy concentrations of diabetic ApoE-/-mice were significantly increased,and the body weight was significantly reduced.The concentrations of blood lipids(TG,LDL,TC),blood glucose,and Hcy can inhibit the occurrence of vascular endothelial glucose and lipid metabolism disorders and effectively reduce the risk factors of atherosclerosis.3.Diabetic state of ApoE-/-mice,reduce the ratio of type Ⅰ/Ⅲ collagen and collagen content in atherosclerotic plaque,reduce the stability of plaque;allicin intervention can increase in atherosclerotic plaque Type Ⅰ/Ⅲcollagen ratio and collagen content reduce the vulnerability of atherosclerotic plaques,stabilize unstable atherosclerotic plaques,and have a dose-effect relationship.4.The expression levels of IL-6 and MMP-9 in diabetic ApoE-/-mice were significantly increased.Allicin intervention can effectively inhibit the activity and expression of IL-6 and MMP-9. |
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