title: A vital role for Angptl3 in the PAN-induced podocyte loss by affecting detachment and apoptosis in vitro.
abstract: BACKGROUND
  Podocyte detachment and apoptosis are two risk factors causing podocyte loss, F-actin rearrangement is involved in detachment and apoptosis. However, the nature of events that promote detachment and apoptosis of podocytes and whether detachment occurred simultaneously with apoptosis are still unclear. Previously, it was found that angiopoietin-like3 (Angptl3) induces F-actin rearrangement in podocytes. In this study we investigate whether Angptl3 influences podocyte loss (detachment and apoptosis) and the process through which Angptl3 exactly influenced the podocyte loss.
  
  
  METHODS
  In conditionally immortalized mice podocytes, recombinant mice Angptl3 protein (rm-Angptl3) was used to mimic Angptl3 overexpression model and transfection with small interfering RNA (siRNA) to knockdown the expression of Angptl3. Both flow cytometry analysis and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay were used to detect apoptosis. Rearrangement of F-actin was assessed using confocal microscopy. Western blot assay was used to measure levels of Angptl3, integrin α3β1, integrin-linked kinase (ILK), p53, caspase 3, and phosphorylation of integrin β1.
  
  
  RESULTS
  In a puromycin aminonucleoside (PAN)-induced podocyte injury model, rm-Angptl3 accelerated the loss of podocytes, both detachment and apoptosis occurred, and F-actin rearrangement is involved in the process. However, knockdown of Angptl3 by siRNA markedly ameliorated these injuries. Observed effects were partially correlated with the altered integrin α3β1, ILK and p53, rather than caspase 3.
  
  
  CONCLUSIONS
  Angptl3 is a novel factor involved in the PAN-induced podocyte loss by affecting detachment and apoptosis in vitro. This study helps to deepen the understanding of the mechanisms of podocyte loss and lays the foundation for developing a new successful therapy for podocyte injury via lower expression of Angptl3.
authors.affiliation: Department of Nephrology and Rheumatism, Children's Hospital of Fudan University, Shanghai, 201102, China. dairufeng2012@163.com.
authors.forename: Rufeng
authors.initials: R
authors.lastname: Dai
authors.affiliation: Department of Pediatrics, Affiliated Hospital of Qingdao University, Shandong, 266003, China. qylinyi@163.com.
authors.forename: Yi
authors.initials: Y
authors.lastname: Lin
authors.affiliation: Department of Nephrology and Rheumatism, Children's Hospital of Fudan University, Shanghai, 201102, China. liu__hm@163.com.
authors.forename: Haimei
authors.initials: H
authors.lastname: Liu
authors.affiliation: Department of Nephrology and Rheumatism, Children's Hospital of Fudan University, Shanghai, 201102, China. amayraojia@gmail.com.
authors.forename: Jia
authors.initials: J
authors.lastname: Rao
authors.affiliation: Department of Nephrology and Rheumatism, Children's Hospital of Fudan University, Shanghai, 201102, China. marinezyh@163.com.
authors.forename: Yihui
authors.initials: Y
authors.lastname: Zhai
authors.affiliation: Department of Biochemistry and Molecular Biology, Shanghai Medical College, Fudan University, Shanghai, 200032, China. xlzha@shmu.edu.cn.
authors.forename: Xiliang
authors.initials: X
authors.lastname: Zha
authors.affiliation: Department of Nephrology and Rheumatism, Children's Hospital of Fudan University, Shanghai, 201102, China. ffxy@vip.sina.com.
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: Fang
authors.affiliation: Department of Nephrology and Rheumatism, Children's Hospital of Fudan University, Shanghai, 201102, China. hxu@shmu.edu.cn.
authors.forename: Hong
authors.initials: H
authors.lastname: Xu
mesh_terms: D000091303:Angiopoietin-Like Protein 3; D000075802:Angiopoietin-like Proteins; D042682:Angiopoietins / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D015415:Biomarkers / Q000032:analysis; D015153:Blotting, Western; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry / Q000379:methods; D005455:Fluorescent Antibody Technique; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D050199:Podocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D011692:Puromycin Aminonucleoside / Q000494:pharmacology*; D034741:RNA, Small Interfering / Q000378:metabolism; D012380:Role; D012680:Sensitivity and Specificity; D014162:Transfection
chemical_list: D000091303:Angiopoietin-Like Protein 3; D000075802:Angiopoietin-like Proteins; D042682:Angiopoietins; C404495:Angptl3 protein, mouse; D015415:Biomarkers; D034741:RNA, Small Interfering; D011692:Puromycin Aminonucleoside