Expression and activation of STAT3 in chronic proliferative immune complex glomerulonephritis and the effect of fosinopril.
BACKGROUND
Signal transducers and activators of transcription (STATs) are cytoplasmic proteins that are activated in response to stimulation from various cytokines. Among these, STAT3 is an important member that has been implicated in the inflammatory proliferation of cells. We hypothesized that STAT3 may be activated in kidneys of rats having modified chronic immune complex glomerulonephritis, and that angiotensin-converting enzyme (ACE) inhibition with fosinopril may prevent the activation of STAT3 and subsequent upregulation of tissue inhibitor of metalloproteinase-1 (TIMP-1), which are effects that may explain the therapeutic effects of fosinopril on nephritis.


METHODS
Fifty-one Wistar rats were randomly divided into three groups that included a control group, a model group and a fosinopril group. Bovine serum albumin (BSA) nephritis was induced by subcutaneous immunization and daily intraperitoneal (i.p.) administration of BSA. To accentuate the nephritis, we performed uni-nephrectomy and gave 100 microg of lipopolysaccharide (LPS) as an i.p. injection. Macrophage infiltration (ED-1) was assessed with immunohistochemistry. The expression and activation of STAT3 and the expression of TIMP-1, one of the STAT3 downstream genes, were observed in renal tissues of rats by means of immunohistochemistry, electrophoretic mobility shift assay (EMSA), western blot and northern blot. The relationships between STAT3 phosphorylation, 24 h urinary protein excretion and TIMP-1 expression were also analysed.


RESULTS
Northern blot showed that the mRNA expression of both STAT3 and TIMP-1 was significantly increased in kidneys from the model group, but significantly decreased in the fosinopril group (P<0.05). Western blot analysis revealed similar increases in the expression of STAT3, phospho-STAT3 (p-STAT3) and TIMP-1 in the model group. Analysis of immunohistochemistry showed that STAT3 and p-STAT3 were expressed in very few cells of normal rats, that expression was strong in model rats and that this increased expression was attenuated in the fosinopril group (P<0.05). The expression of p-STAT3 in glomeruli was positively correlated with 24 h proteinuria as well as with glomerular TIMP-1 expression. Double staining showed that some ED-1-positive cells also contained p-STAT3-positive staining.


CONCLUSIONS
The present study showed that STAT3 is expressed and activated in kidneys of rats with modified immune complex glomerulonephritis. These rats also had increased ED-1-positive cells, with some cells showing simultaneous expression of p-STAT3 and ED-1, which may contribute to glomerular inflammatory proliferation and extracellular matrix accumulation. Finally, fosinopril downregulated STAT3 activation and ED-1 influx, which are effects that may attenuate renal damage in this model.
Zhang
Wuxing
W
Department of Nephrology, General Hospital of PLA, Beijing 28 Fuxing Road, 100853, P.R. China.
Chen
Xiangmei
X
Shi
Suozhu
S
Wei
Ribao
R
Wang
Jianzhong
J
Yamanaka
Nobuaki
N
Hong
Quan
Q
D000806:Angiotensin-Converting Enzyme Inhibitors / Q000494:pharmacology*; D000818:Animals; D000936:Antigen-Antibody Complex / Q000276:immunology*; D001794:Blood Pressure; D002908:Chronic Disease; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000032:analysis* / Q000378:metabolism; D005260:Female; D017328:Fosinopril / Q000494:pharmacology*; D015432:Glomerulonephritis, Membranoproliferative / Q000378:metabolism*; D007150:Immunohistochemistry; D007668:Kidney / Q000473:pathology; D010766:Phosphorylation; D011507:Proteinuria / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D050796:STAT3 Transcription Factor; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000032:analysis; D015534:Trans-Activators / Q000032:analysis* / Q000378:metabolism
D000806:Angiotensin-Converting Enzyme Inhibitors; D000936:Antigen-Antibody Complex; D004268:DNA-Binding Proteins; D050796:STAT3 Transcription Factor; C494088:Stat3 protein, rat; D019715:Tissue Inhibitor of Metalloproteinase-1; D015534:Trans-Activators; D004247:DNA; D017328:Fosinopril