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. 2009 Feb;58(2):469-77.
doi: 10.2337/db08-1328. Epub 2008 Nov 18.

Enhanced expression of Janus kinase-signal transducer and activator of transcription pathway members in human diabetic nephropathy

Celine C Berthier  1 , Hongyu Zhang, MaryLee Schin, Anna Henger, Robert G Nelson, Berne Yee, Anissa Boucherot, Matthias A Neusser, Clemens D Cohen, Christin Carter-Su, Lawrence S Argetsinger, Maria P Rastaldi, Frank C Brosius, Matthias Kretzler
Affiliations

Enhanced expression of Janus kinase-signal transducer and activator of transcription pathway members in human diabetic nephropathy

Celine C Berthier et al. Diabetes. 2009 Feb.

Abstract

Objective: Glomerular mesangial expansion and podocyte loss are important early features of diabetic nephropathy, whereas tubulointerstitial injury and fibrosis are critical for progression of diabetic nephropathy to kidney failure. Therefore, we analyzed the expression of genes in glomeruli and tubulointerstitium in kidney biopsies from diabetic nephropathy patients to identify pathways that may be activated in humans but not in murine models of diabetic nephropathy that fail to progress to glomerulosclerosis, tubulointerstitial fibrosis, and kidney failure.

Research design and methods: Kidney biopsies were obtained from 74 patients (control subjects, early and progressive type 2 diabetic nephropathy). Glomerular and tubulointerstitial mRNAs were microarrayed, followed by bioinformatics analyses. Gene expression changes were confirmed by real-time RT-PCR and immunohistological staining. Samples from db/db C57BLKS and streptozotocin-induced DBA/2J mice, commonly studied murine models of diabetic nephropathy, were analyzed.

Results: In human glomeruli and tubulointerstitial samples, the Janus kinase (Jak)-signal transducer and activator of transcription (Stat) pathway was highly and significantly regulated. Jak-1, -2, and -3 as well as Stat-1 and -3 were expressed at higher levels in patients with diabetic nephropathy than in control subjects. The estimated glomerular filtration rate significantly correlated with tubulointerstitial Jak-1, -2, and -3 and Stat-1 expression (R(2) = 0.30-0.44). Immunohistochemistry found strong Jak-2 staining in glomerular and tubulointerstitial compartments in diabetic nephropathy compared with control subjects. In contrast, there was little or no increase in expression of Jak/Stat genes in the db/db C57BLKS or diabetic DBA/2J mice.

Conclusions: These data suggest a direct relationship between tubulointerstitial Jak/Stat expression and progression of kidney failure in patients with type 2 diabetic nephropathy and distinguish progressive human diabetic nephropathy from nonprogressive murine diabetic nephropathy.

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Figures

FIG. 1.
FIG. 1.
Jak/Stat canonical pathway in human diabetic nephropathy as assessed by Ingenuity Pathway Analysis software. The Jak/Stat family members represented include Jak-1, -2, and -3 and Stat-1, -2, -3, -4, -5A, -5B, and -6. The mRNA expression changes for Jak/Stat family members in early (PIMA Indians, n = 22–24) (left panel) and progressive (European, n = 7–11) (right panel) human diabetic nephropathy (DN) are indicated in red for increased expression and green for decreased expression, compared with the corresponding control group (LD + MCD, n = 7–12). 2000–2008 Ingenuity Systems. All rights reserved. LD, living donor; MCD, minimal change disease.
FIG. 2.
FIG. 2.
Jak-2 mRNA expression levels in human diabetic nephropathy: correlation with renal function. A: Increased expression of Jak-2 mRNA in the glomeruli of early diabetic nephropathy patients and in the tubulointerstitium of progressive diabetic nephropathy patients compared with control subjects (n = 6–9 in the control group, n = 11–12 in the early diabetic nephropathy group, and n = 10–12 in the progressive diabetic nephropathy group). □, control; formula image, early diabetic nephropathy; ▪, progressive diabetic nephropathy. B: Correlation of tubulointerstitial Jak-2 mRNA expression (as measured by real-time RT-PCR) with eGFR in patients with early (×, n = 11) and progressive (•, n = 12) diabetic nephropathy. C: Representative Jak-2 immunohistographs of kidney biopsies from patients with no kidney disease (Control), progressive diabetic nephropathy (Prog. DN), hypertensive nephropathy (HTN), IgA nephropathy (IgAN), or lupus nephritis (LN). Jak-2 expression was substantially and significantly increased in the proximal tubular cells and glomeruli of the patients with diabetic nephropathy compared with the control and not in other progressive kidney diseases. (Please see http://dx.doi.org/10.2337/db08-1328 for a high-quality digital representation of this figure.)
FIG. 3.
FIG. 3.
Jak-2 in the C57BLKS type 2 diabetic nephropathy and STZ-induced DBA/2J type 1 diabetic nephropathy mouse models. A: Jak-2 real-time RT-PCR mRNA expression in cortex (n = 6 control, n = 7 diabetic) and glomeruli (n = 5 control, n = 7 diabetic) was not altered in db/db mice (▪) compared with db/m control animals (□). B: Jak-2 immunoblot (upper panel) and densitometry (lower panel). Jak-2 protein levels were unchanged in the cortex (n = 3 control, n = 5 diabetic) and glomeruli (n = 4 in both groups) of db/db mice (▪) versus db/m mice (□). C: Jak-2 qRT-PCR mRNA expression in cortex and glomeruli was not altered in DBA/2J diabetic mice (▪) compared with control animals (□) (n = 5 in each group). D: Jak2 immunoblot (upper panel) and densitometry (lower panel) in DBA/2J mice. ▪, diabetic; □, control. Jak-2 protein levels were not significantly different in the cortex or glomeruli of DBA/2J diabetic mice. n.s., statistically nonsignificant.
FIG. 4.
FIG. 4.
Effects of Jak-2 overexpression and high glucose on Stat-3 phosphorylation in Jak-2 mesangial cells. A: Jak-2 (130 kDa), total Stat-3 (79 kDa), phosphorylated Stat-3 (79 kDa), and β-tubulin (50 kDa) immunoblots of murine mesangial cells transfected with a Jak-2 prk-5 plasmid or a control vector (n = 3 per group). B: Phosphorylated Stat-3 levels normalized to total Stat-3 levels.
FIG. 5.
FIG. 5.
Effects of Jak-2 overexpression and high glucose on ROS generation in mesangial cells. A: DCF fluorescence in murine mesangial cells transfected with either Jak-2 prk-5 or control vectors exposed to 5.5 mmol/l d-glucose + 24.5 mmol/l mannitol or 30 mmol/l d-glucose for 24 h. Image exposure times were identical and are representative of those from three separate fields on five separate slides for each condition. B: ROS quantitation from the DCF fluorescence experiments. Fluorescence intensity was determined using NIH ImageJ. (n = 5 per group; 50 cells were assessed for each slide).
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References

    1. Mauer SM, Steffes MW, Ellis EN, Sutherland DE, Brown DM, Goetz FC: Structural-functional relationships in diabetic nephropathy. J Clin Invest 74: 1143–1155, 1984 - PMC - PubMed
    1. Mauer SM, Lane P, Zhu D, Fioretto P, Steffes MW: Renal structure and function in insulin-dependent diabetes mellitus in man. J Hypertens Suppl 10: S17–20, 1992 - PubMed
    1. Chavers BM, Bilous RW, Ellis EN, Steffes MW, Mauer SM: Glomerular lesions and urinary albumin excretion in type I diabetes without overt proteinuria. N Engl J Med 320: 966–970, 1989 - PubMed
    1. Nangaku M: Mechanisms of tubulointerstitial injury in the kidney: final common pathways to end-stage renal failure. J Int Med 43: 9–17, 2004 - PubMed
    1. Bader R, Bader H, Grund KE, Mackensen-Haen S, Christ H, Bohle A: Structure and function of the kidney in diabetic glomerulosclerosis: correlations between morphological and functional parameters. Pathol Res Pract 167: 204–216, 1980 - PubMed

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