Effect Of Sulodexide On The Change Of MMP Expression And Autophagy Activity In Aortic Medial Calcification In Uremic Rats

Background:Vascular calcification is a major complication of uremic patients,as well as the high incidence and high mortality of cardiovascular disease.Unlike atherosclerosis caused by endometrial vascular calcification,uremia patients with vascular calcification happens at the media of Large and medium sized arteries,which can lead to increased arterial stiffness,increased pulse pressure.The mechanism of vascular calcification in uremia is not fully clear.Studies have shown that increased expression of matrix metalloproteinases(MMPS)and vascular smooth muscle cells(VSMC)autophagy activity impaired in patients promote arterial calcification process.Sulodexide(SDX)have significant resistance to artery intimal hyperplasia,inhibits MMP expression.So far,did not see whether SDX can effect media of Large arteries calcification or not in research reports.Objective:The observation of uremic rats aorta vascular wall MMPS expression and change of the autophagy activity,As well as the two relations with aortic vascular calcification;To observe the influence of Sulodexide for uremic rats aorta vascular wall of MMPS expression,autophagy activity and vascular calcification.Method:30 male Sprague wrote-Dawley rats,random selection table number 10 to normal diets,as a healthy control group,the remaining 20 to 0.75% adenine higher phosphorus diet(1%),to make uremia animal model.In 4 weeks,20 weeks to collect blood specimens,detecting of blood serum creatinine,calcium,phosphorus,as the quantitative biochemical indicators.At the 20 weekend,we put the rats to death,take the abdominal aorta,part of the line organization fixed of paraffin section,HE,Masson and von Kossa staining,observed the pathological changes and the aorta vascular calcification,immune histochemical method to detect the core application combined with factor alpha 1(Cbfɑ-1)and osteopontin(OPN)in the aorta of positioning and expression;Line of frozen tissue in situ dyed gelatin enzymes,observation of MMPS expression,parallel immunofluorescence staining,observed LC3 distribution.Some tissue cryopreservation,homogenate after Western imprinting testing,observing aortic vascular wall OPN,Cbfɑ-1 and autophagy related molecular LC3,p62 protein expression.Part of the organization after glutaraldehyde fixation,transmission electron microscopy examination.Result:4 weeks after building,uremia,SDX group rats blood serum BUN,Scr and phosphorus were significantly higher than that of control group(all P < 0.01);Building after 20 weeks,uremia and SDX group rat aorta markedly thickened,membrane calcification in the visible,but SDX aortic calcification,to a lesser degree(P < 0.05);Immunohistochemistry showed that compared with control group,uremia,SDX membrane and outer membrane in rat aortic endothelial,MMP2,MMP9,OPN,increase Cbf alpha 1 expression,but SDX express the uremia group decreased significantly(P < 0.05);Gelatin in situ enzyme spectrum analysis found that compared with control group,uremia,SDX group rat aorta MMP activity increased significantly,but SDX express the uremia group decreased significantly(P < 0.05);Transmission electron microscopic examination showed that the control membrane in the aortic smooth muscle cells exist obvious autophagy,uremia,SDX autophagy bubble number less obvious.Immunofluorescence showed that compared with control group,uremia,SDX group rat aorta LC3 expression decreased,but the SDX express the uremia group have obvious recovery(P < 0.05).Western mark shows that compared with the control group,uremia,SDX group rats increased aortic p62 expression,LC3-Ⅱ/LC3-Ⅰ ratio drops,OPN,Cbf alpha 1 expression increased(all P < 0.05),but the SDX express the uremia group have obvious recovery(all P < 0.05);Correlation analysis found that aortic vascular wall of MMPS expression and p62 expression were positively correlated(r = 0.32,P < 0.05),and LC3-Ⅱ/LC3-Ⅰ expression showed a negative correlation(r = 0.51,P < 0.05);Aortic vascular wall LC3-Ⅱ/LC3-Ⅰ expression and OPN,Cbf alpha 1 expression showed a negative correlation(r = 0.72,0.66,P < 0.05),p62 expression and OPN,Cbf alpha 1 expression was positively correlated(r = 0.46,0.60,P < 0.05).Conclusion:MMP expression in uremic rats aorta vascular wall increases significantly,and autophagy activity significantly decreases,both are closely associated with aortic vascular calcification;Sulodexide can inhibit the activation of MMP in aortic vascular wall,and regulating autophagy activity of aortic smooth muscle cells,so as to alleviates vascular calcification of uremic rats aorta.