Inhibiting Effects Of Calycosin On Expression Of ICAM-1 In Vascular Endothelial Cell And Its Receptor LFA-1

Background:Vascular endothelial cells and its function have more relationship with chronic kidney diseases, its dysfunction contributes to the progression of CKD, finally results into end-stage kidney disease, and contributes substantially to morbidity and mortality, Causing serious social and economic burden. Cytokines by promoting the ICAM-1 secretion and its receptor expression of LFA-1 to promote the inflammatory cascade, inflammation causes VEC injury, dysfunction, thus contributing to progression in CKD. ICAM-1 and its receptor LFA-1 interaction is the start-factor of development of inflammation. Therefore, to explore drugs, which slow the progression of CKD, has important social, economic significance.Objective:To investigate effects of Astragalus monomer calycosin on the expression of ICAM-1 in vascular endothelial cell and its receptor LFA-1, For clinical treatment and provide the basis for drug development, In vitro cultured ECV-304 cells were studied, and health human PMN were incubated, tumor necrosis factor a was given to stimulate the cells.Methodology:ECV-304 cells were derived from CCTCC, the cells were cultured in vitro with L-DMEM and 10% fetal bovine serum medium in 5%CO2,37℃, Saturated humidity to sufficient numbers. When the cells grew to fusion of 80%-90%, adjusted cell numbers to 2×105/ml after being washed with PBS and digested by trypsin, then cultured in L-DMEM with 10% fetal bovine serum for 48h and without fetal bovine serum for 12h,Then ECV304 cells were established and divided into control groups (not joining any intervening factors) and TNF a groups(40ng/ml) and test groups of calycosin at different concentration (10,1,0.1mg/l) randomly. Before TNFαinjection treatment, Added PMN suspension 100ul per well in each group, which derived from healthy volunteers and cultured with L-DMEM medium at 1:4-volume ratio. ELISA on the condition of 450nm wavelength detected the production of both ICAM-1 and LFA-1; there were 10 duplicate wells in each group. The expression level of ICAM-1 and LFA-1 has a positive correlation with OD450 value. Completely random design one-way ANOVA statistical method was used to compare and analyze the results. The value of OD450 of all groups were expressed in Mean±SD, SNK test was used for multiple comparison, LSD method was used for comparison between groups, p<0.05 meant having statistical significance.Results:1) Compared with controls,TNFα(40ng/ml) promoted the synthesis of both ICAM-1 and LFA-1 (p<0.05);2) Compared with TNFαgroups, all of the calycosin groups inhibited the production of ICAM-1 and LFA-1 (p<0.05);3) Compared between 3 calycosin groups, the highest concentration calycosin groups could significantly decrease the expression of both ICAM-1 and LFA-1 p<0.05).Conclusion:Calycosin can inhibit ICAM-1 over-expression in ECV-304 cells and LFA-1 on PMN induced by the stimulation of TNFα.Its effects were dose-dependently. Calycosin can protect endothelial cells.