Up-regulation of HSP47 in the mouse kidneys with unilateral ureteral obstruction.
BACKGROUND
Unilateral ureteral obstruction (UUO) is a well established experimental model of renal injury leading to interstitial fibrosis. The molecular and cellular mechanism(s) of interstitial fibrosis in UUO are beginning to be elucidated. In the progression of interstitial fibrosis in UUO, up-regulation of collagen synthesis is commonly observed. HSP47 is a collagen-binding stress protein and is thought to be a collagen-specific molecular chaperone, which plays a pivotal role during the biosynthesis and secretion of collagen molecules in the endoplasmic reticulum. The synthesis of HSP47 has been demonstrated to always parallel that of collagen in physiological and pathophysiological conditions. It is well recognized that renin-angiotensin system (RAS) is enhanced in the setting of UUO and that enhanced RAS has been implicated in the pathogenesis of interstitial fibrosis in the obstructed kidneys.


METHODS
To investigate the role of HSP47 in the progression of interstitial fibrosis in mouse UUO, the expression of HSP47 was examined by Northern blotting, immunohistochemistry and in situ hybridization in the obstructed kidneys. To test the possible involvement of enhanced RAS on the HSP47 expression, we examined the effects of lisinopril, an angiotensin converting enzyme inhibitor, on interstitial fibrosis. HSP47 and type I collagen mRNA expression.


RESULTS
By Northern blot analysis, HSP47 mRNA was significantly up-regulated at 12 hours (about twice that of sham operated kidneys) after the onset of ureteral obstruction, further increased and stayed at the increased level until seven days (about 8 times that of sham operated kidneys). HSP47 mRNA and protein expression were observed in the periglomerular and peritubular interstitial regions of the obstructed kidneys. Distribution of smooth muscle alpha actin and type I collagen immunoreactivity were similar to the HSP47 distribution pattern, suggesting that HSP47 was up-regulated in the myofibroblasts. Lisinopril ameliorated the expansion of cortical interstitium in the obstructed kidneys at four and seven days after ureteral obstruction. HSP47 mRNA expression was suppressed at four and seven days, whereas type I collagen mRNA was suppressed only at seven days after the onset of ureteral obstruction.


CONCLUSIONS
These results demonstrate the early and persistent up-regulation of HSP47 during the progression of interstitial fibrosis in mouse UUO kidneys, and further suggest the potential role of HSP47 in the pathogenesis of interstitial fibrosis in the obstructed kidneys. Partial suppression of HSP47 mRNA expression by lisinopril at day 4 and day 7 after ureteral obstruction suggests that there are other immediate trigger(s) that induce the HSP47 mRNA expression. Identification of the molecular mechanism of HSP47 induction during UUO may give an insight into the novel aspects of the molecular pathophysiology of interstitial fibrosis in obstructive nephropathy.
Moriyama
T
T
First Department of Medicine, Osaka University School of Medicine, Faculty of Health and Sport Sciences, Japan. toshiki@medone.med.osaka-u.ac.jp
Kawada
N
N
Ando
A
A
Yamauchi
A
A
Horio
M
M
Nagata
K
K
Imai
E
E
Hori
M
M
D000199:Actins / Q000032:analysis; D000806:Angiotensin-Converting Enzyme Inhibitors / Q000494:pharmacology; D000818:Animals; D003094:Collagen / Q000235:genetics; D004195:Disease Models, Animal; D005355:Fibrosis; D015870:Gene Expression / Q000187:drug effects; D050958:HSP47 Heat-Shock Proteins; D006360:Heat-Shock Proteins / Q000032:analysis / Q000235:genetics*; D007124:Immunoenzyme Techniques; D017403:In Situ Hybridization; D007668:Kidney / Q000737:chemistry / Q000378:metabolism* / Q000473:pathology; D017706:Lisinopril / Q000494:pharmacology; D008297:Male; D051379:Mice; D008809:Mice, Inbred C3H; D012333:RNA, Messenger / Q000032:analysis; D012083:Renin / Q000097:blood; D012084:Renin-Angiotensin System / Q000502:physiology; D014517:Ureteral Obstruction / Q000188:drug therapy / Q000473:pathology / Q000503:physiopathology*
D000199:Actins; D000806:Angiotensin-Converting Enzyme Inhibitors; D050958:HSP47 Heat-Shock Proteins; D006360:Heat-Shock Proteins; D012333:RNA, Messenger; C494661:Serpinh1 protein, mouse; D003094:Collagen; D017706:Lisinopril; D012083:Renin