Discussion On The Etiology And Molecular Mechanism Of Major Cardiovascular Diseases In Patients With Chronic Kidney Disease From The Perspective Of SCAP Dysfunction

PART1 Establishment of vascular calcification model of umbilical vein in vitro with high phosphorus cultureObjective: To establish an arterial hyperphosphoric culture model to simulate the process of vascular calcification(VC)during hyperphosphatemia in vivo.Methods: Umbilical vein M199 medium was cultured in serum without serum,divided into normal group(Pi = 1mmol / L)and high phosphorus group(Pi = 3mmol / L).The umbilical vein sections were stained by Von Kossa staining and alizarin red staining.The advantages and disadvantages of the two methods were observed and compared.The morphology of umbilical vein was evaluated by HE staining.Results: In vitro vascular calcification model was successfully modeled,and calcification in high phosphorus group was significantly higher than that in normal group(P <0.01).In vitro culture in the twentieth day when the organizational structure is clear,clear staining of the nucleus,cytoplasm red dye.Comparison of two kinds of calcification method,Von kossa staining method for the middle layer of microcalcification than the alizarin red clearer.Conclusion: Von kossa staining was used to identify arterial microcalcification in 20 days of in vitro high phosphorus(Pi = 3mmol / L)culture.PART 2 The interrelationship between calcification and SCAP expression in arterial medial vascular smooth muscle cellsObjective: To study the effect of SCAP protein on the deposition of calcium salt in the model of hyperphosphoric artery calcification in vitro by lentiviral knockout of umbilical vein SCAP.Methods: The umbilical vein was removed from the maternal serum for 2 days,and the virus was cultured for 3 days.The rats were divided into four groups: control group,high phosphorus group,SCAP knockout group,high phosphorus + SCAP Knockout group.VK staining was used to quantify vascular calcification and WB to detect the expression of SCAP knockout and RUNX2 in four groups.Results: Von kossa staining and quantitative calculation of calcium tube showed that the high-phosphorus + SCAP protein knockout group was higher than that of SCAP protein knockout group(P <0.01)The expression of RUNX2 protein in high phosphorus group was significantly higher than that in control group(P <0.01).The expression of RUNX2 protein in high phosphorus + SCAP knockout group was significantly lower than that in SCAP knockout group(P <0.05).Conclusion: SCAP protein may be involved in the deposition of calcium phosphate in the middle of the phosphorus and promote the calcification.PART 3 serum phosphorus by regulating SCAP protein overexpression to accelerate the process of atherosclerosisObjective: To investigate the effect of phosphorus on atherosclerosis in patients with chronic kidney disease(CKD)by dialectomy,radial artery biopsy and umbilical vein in vitro.Methods: CKD cases were divided into atherosclerosis(AS)group and non-atherosclerosis(NAS)group.Logistic regression analysis was used to test the difference between the two groups.(SREBP Cleavage Activating Protein(SCAP)),LDL receptor(LDL receptor),cholesterol(LDL receptor),and LDL receptor(LDL receptor)were measured by immunofluorescence method in the radial artery.(HMGCo AR)was detected by immunohistochemistry and immunohistochemistry.Results The expression of HMGCo AR was detected by immunohistochemistry and immunohistochemistry.Results The expression of HM-Coase Venous SCAP,LDLr,SREBP2,HMGCo AR expression.Results: Logistic regression analysis showed that serum phosphorus was an independent risk factor for AS.The expression of SCAP,EREBP2,HMGCo AR,LDLr and RUNX2 in high phosphorus group were higher than those in normal group(P <0.05).(P <0.05).The results of immunohistochemistry showed that the expression of SCAP,SREBP2,HMGCo A and LDLr were significantly higher than those of the control group(P <0.05).Conclusion: Serum phosphorus may accelerate the development of atherosclerosis by increasing the SCAP protein and breaking the negative feedback regulation of SCAP-SREBP2 pathway,leading to accumulation of lipid in endometrial cells.