Coagulation in the mesangial area promotes ECM accumulation through factor V expression in MsPGN in rats.
It is well known that tissue factor starts the extrinsic coagulation pathway, which activates factor X to Xa, and factor V is a membrane-bound potent cofactor for the terminating stage of prothrombin activation by factor Xa. In a previous in vitro study, factor V was induced in cultured mesangial cells by inflammatory stimulation and increased expression of factor V promoted fibrin generation on the cultured mesangial cell surface. We report that extracellular matrix (ECM) accumulation is increased in association with coagulation in the mesangial area through factor V expression in mesangioproliferative glomerulonephritis (MsPGN). Wistar rats were intravenously injected with rabbit anti-rat thymocyte serum accompanied with or without simultaneous injection of rabbit anti-factor V antibody. Time course study in immunohistochemistry revealed that factor V expression was prominent on day 3 and fibrin-related antigen (FRA) deposition, then ECM accumulation, followed from day 3 to day 8. Massive fibronectin depositions and transforming growth factor (TGF)-beta expression were also noted in glomeruli from the disease control group, markedly higher than those in the normal group, and these depositions and expressions were significantly decreased in the anti-factor V neutralizing antibody-injected group. Northern blot analysis revealed that factor V mRNA expression was prominent on day 3 and was weak on day 8. Double-labeling experiments revealed the frequent colocalization of alpha-smooth muscle actin with factor V, FRA, and fibronectin in the same mesangial areas of glomeruli. TGF-beta, connective tissue growth factor (CTGF), collagen type IV, and fibronectin mRNA were upregulated in the disease control group, and anti-factor V-neutralizing antibody injection suppressed these mRNA expressions in glomeruli. The present results suggest that ECM components accumulation may progress in accordance with coagulation in the mesangial area through mesangial factor V expression and upregulated expression of TGF-beta and CTGF in MsPGN.
Liu
Ning
N
Division of Nephrology, Department of Cardiovascular Medicine, Graduate School of Pharmaceutical Sciences, Kyoto University, Kyoto 606-8507, Japan.
Makino
Toshiaki
T
Nogaki
Fumiaki
F
Kusano
Hitoshi
H
Suyama
Katsuo
K
Muso
Eri
E
Honda
Gisho
G
Kita
Toru
T
Ono
Takahiko
T
D000199:Actins / Q000378:metabolism; D000818:Animals; D000941:Antigens / Q000378:metabolism; D001777:Blood Coagulation / Q000502:physiology*; D015152:Blotting, Northern; D024141:Collagen Type IV / Q000235:genetics / Q000378:metabolism; D055513:Connective Tissue Growth Factor; D005109:Extracellular Matrix / Q000187:drug effects / Q000378:metabolism*; D005165:Factor V / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D005353:Fibronectins / Q000235:genetics / Q000378:metabolism; D005920:Glomerular Mesangium / Q000098:blood supply / Q000378:metabolism*; D005921:Glomerulonephritis / Q000378:metabolism / Q000503:physiopathology*; D006801:Humans; D017874:Immediate-Early Proteins / Q000235:genetics / Q000378:metabolism; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism; D008264:Macrophages / Q000276:immunology; D008297:Male; D019001:Nitric Oxide Synthase / Q000378:metabolism; D052247:Nitric Oxide Synthase Type II; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D011517:Prothrombin Time; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017208:Rats, Wistar; D013917:Thrombin / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000235:genetics / Q000378:metabolism
D000199:Actins; D000941:Antigens; C526317:CCN2 protein, human; C526316:CCN2 protein, rat; D024141:Collagen Type IV; D005353:Fibronectins; D017874:Immediate-Early Proteins; D036341:Intercellular Signaling Peptides and Proteins; D018809:Proliferating Cell Nuclear Antigen; D012333:RNA, Messenger; D016212:Transforming Growth Factor beta; C023456:fibrinogen-related antigen; D055513:Connective Tissue Growth Factor; D005165:Factor V; C496319:NOS2 protein, human; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; C496318:Nos2 protein, rat; D013917:Thrombin