Both PAI-1 and uPA bind to uPAR, and make complex with integrin on cell membrane. Internalization of the complex induces the cell detachment as a result of reduction of cell-matrix adhesion molecules. The present study was aimed to show that PAI-1 was involved in podocyte detachment through the complex with uPAR-integrin by using NEP mice and podocyte cell line. Methods: Two groups of NEP mice, with or without PAI-1 inhibitor (PI) were induced podocyte injury by LMB2 injection on day 0. PI was administered from day 0 to 12. Histological and clinical parameters were analyzed
on day 12. Then, we treated cultured podocytes either with PAI-1/uPA complex (P/U), uPA (control), or antibody for blocking uPAR with P/U (B-P/U). After incubation, attached cells were counted, and localization of β1 integrin and uPAR was detected by immunofluorescence Selleckchem AZD1208 learn more and double immunolabeling electron microscopy. Cytoplasmic β1 integrin was analyzed by Western blot. Results: Proteinuria (P) and Thronbi score (T) in PI group were lower than the control (P;
64.29 ± 23.30 vs. 161.12 ± 34.0; p < 0.05, T; 0.01 ± 0.01 vs. 0.23 ± 0.07, p < 0.05), and podocyte numbers were preserved (9.41 ± 0.45 vs. 2.67 ± 0.41, p < 0.0001). Glomerular morphology in PI group was preserved. In vitro, attached cells in P/U were reduced compared with the control and B-P/U (p < 0.01). Confocal microscopy showed that β1 integrin and uPAR were colocalized (Pearson's coefficient (PC) = 0.50) and shifted to cytoplasm in P/U. In contrast, β1 integrin remained on the membrane and was not colocalized with uPAR in the control and B-P/U (PC = 0.12, 0.06, respectively). In Western blot, β1 integrin expression was increased in P/U. Double immunolabeling electron microscopy showed co-localization of β1 integrin and uPAR in the endocytotic vesicles in podocytes. Conclusion: PAI-1/uPA complex
may act on the podocytes detachment Loperamide via internalization of β1 integrin through the uPAR mechanism. FAN QIULING, LI SALI, LIU NAN, JIANG YI, WANG LINING Department of Nephrology, the First Affiliated Hospital of China Medical University, Shenyang, China Introduction: Analyze the correlation and risk factors between clinical indicators and the four main pathological lesions of the Oxford classification in IgAN. Methods: Clinical and pathological data were collected from 514 patients with biopsy-proven IgA nephropathy who were 18 years or older. Spearman’s coefficient of rank correlation was performed to evaluate associations between the Oxford classification of IgAN and various clinical indicators. The independent risk factors affecting the pathological classification were analyzed by multivariate regression. Results: The average age of 514 IgAN patients was 35.70 ± 11.99, and the average disease duration was 18.31 ± 30.42 months.