This Article Abstract Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Fujihara, C. K. Articles by Zatz, R. Search for Related Content PubMed PubMed Citation Articles by Fujihara, C. K. Articles by Zatz, R. Related Collections Animal models of human disease Hypertension - basic studies Endothelium/vascular type/nitric oxide

(Hypertension. 2001;37:170.)
© 2001 American Heart Association, Inc.

Scientific Contributions

Mycophenolate Mofetil Reduces Renal Injury in the Chronic Nitric Oxide Synthase Inhibition Model

Clarice Kazue Fujihara; Denise Maria Avancini Costa Malheiros; Irene de Lourdes Noronha; Gilberto De Nucci; Roberto Zatz

From the Renal Division, Department of Clinical Medicine, Faculty of Medicine, University of São Paulo, São Paulo, Brazil.

Correspondence to Roberto Zatz, MD, PhD, Laboratório de Fisiopatologia Renal, Av. Dr. Arnaldo, 455, 3-s/3342, 01246–903, São Paulo, SP, Brazil. E-mail rzatz{at}usp.br

	Abstract

Top Abstract Introduction Methods Results Discussion References

 
Abstract—We and others have recently shown that mycophenolate mofetil (MMF) reduces renal inflammation and glomerular and interstitial injury in the 5/6 renal ablation model. In the present study, we investigated whether MMF limits renal injury in a model of chronic nitric oxide (NO) inhibition associated with a high-salt diet and characterized by progressive systemic hypertension, albuminuria, glomerular sclerosis and ischemia, interstitial expansion, and progressive macrophage infiltration. Adult male Münich-Wistar rats were distributed among 3 groups: HS, rats receiving a high-salt diet (3.2% Na); HS+N, HS rats orally treated with the NO inhibitor N-nitro-L-arginine methyl ester (L-NAME), 25 mg · kg^-1 · d^-1; and HS+N+MMF, HS+N rats orally treated with MMF, 10 mg · kg^-1 · d^-1. Renal hemodynamics were studied after 15 days of treatment; histological and immunohistochemical studies were conducted after 30 days of treatment. MMF treatment did not reverse the hemodynamic alterations characteristic of this model. Renal injury in the HS+N group was associated with macrophage and lymphocyte infiltration. Treatment with MMF reduced glomerular and interstitial injury and limited macrophage and lymphocyte infiltration. These results suggest that renal inflammation is a strong independent factor in the pathogenesis of the nephropathy associated with the HS+N model.

Key Words: mycophenolate mofetil • nitric oxide • renal inflammation • renal hemodynamics

	Introduction

Top Abstract Introduction Methods Results Discussion References

 
Chronic inhibition of NO synthesis by L-arginine analogues, such as N -nitro- L-arginine methyl ester (L-NAME), promotes progressive arterial hypertension associated with proteinuria and severe renal vascular, glomerular, and interstitial injury.^{1 2} These events are exacerbated by the concomitant administration of a high-sodium diet (HS).^{3 4} The pathogenesis of renal injury in rats receiving HS and L-NAME (HS+N model) has not been clarified. Hemodynamic factors such as glomerular hypertension are likely to play a role because treatment with drugs that lower glomerular pressure ameliorates renal injury in this model.^{1 3 5} On the other hand, there is growing evidence that renal inflammation participates in the pathogenesis of renal injury in immune-mediated and nonimmune-mediated progressive nephropathies.^{6 7} NO inhibition enhances leukocyte adhesion and migration,^{8 9} the expression of adhesion molecules,^{9 10} and type I collagen.^{11 12} Accordingly, NO inhibition stimulates cell proliferation and macrophage infiltration.¹³ Thus chronic NO inhibition would be expected to favor the development of renal inflammation. We recently reported preliminary evidence that renal injury in HS+N rats is accompanied by a marked macrophage infiltration and renal cell proliferation.¹⁴

We and others have shown that the immunosuppressor mycophenolate mofetil (MMF), used to prevent allograft rejection,^{15 16} reduces renal lymphocyte and macrophage infiltration and attenuates renal injury in rats with 5/6 renal ablation, a nonimmune-mediated model of progressive nephropathy.^{17 18} In the present study, we investigated whether MMF can similarly reduce inflammation and ameliorate renal injury in HS+N rats.

	Methods

Top Abstract Introduction Methods Results Discussion References

 
Experimental Groups
Fifty-two adult male Münich-Wistar rats, obtained from an established colony at the University of São Paulo, Brazil, and weighing initially 240 to 270 g, were used in this study. The animals were maintained at 23±1°C on a 12/12 hours light/dark cycle, with free access to tap water, and received HS (3.2% Na, Harland Teklad) for 2 weeks. The rats were then divided into 3 groups: Group HS, receiving HS and no drug treatment; Group HS+N, receiving HS and L-NAME (Sigma Co), 0.5 mmol/L dissolved in drinking water, corresponding to a daily intake of 25 mg/kg; and Group HS+N+MMF, receiving HS, L-NAME, and MMF (Roche Laboratories), 10 mg · kg^-1 · d^-1. MMF was dissolved in a mixture of dimethylsulfoxide and olive oil, the final concentration of dimethylsulfoxide being 5%. The compound was administered by gavage once daily in a volume of vehicle never exceeding 0.3 mL. The HS and HS+N groups received vehicle only. All experimental procedures were conducted in accordance with our institutional guidelines.

Renal Hemodynamic Studies
Fifteen days after the beginning of treatments, 4 HS rats, 5 HS+N rats, and 5 HS+N+MMF rats were subjected to renal hemodynamic studies after being anesthetized with Inactin, 100 mg/kg IP, and placed on a temperature-regulated surgical table. Femoral arterial pressure was continuously monitored with a computerized data acquisition system. Saline solution containing ¹⁴C-tagged inulin (0.3 µCi/mL) and homologous rat plasma was infused through the jugular veins. Urine was collected from the left ureter for measurement of flow rate and inulin concentration. Blood samples were obtained from the left renal vein with a sharpened glass micropipette. Hydraulic pressures in superficial glomeruli (P_GC), tubules (P_T), and efferent arterioles (P_E) were measured with a servo-nulling device. Inulin extraction (equivalent to filtration fraction), glomerular filtration rate, renal plasma flow, and renal vascular resistance were calculated with standard equations. Further details of the methods used in these hemodynamic studies are given elsewhere.¹⁹

Long-Term Studies
Twelve HS, 13 HS+N rats, and 13 HS+N+MMF rats were followed-up for 30 days of treatment. At this time, the tail-cuff pressure was measured by an indirect method,¹⁷ and rats were placed in metabolic cages for determination of 24-hour urinary albumin excretion rate (U_albV) by radial immunodiffusion.¹⁷ Rats were thereafter anesthetized with sodium pentobarbital, 50 mg/kg IP, and blood was collected from the abdominal aorta for determination of plasma creatinine concentration (P_creat). The kidneys were then perfusion-fixed at the measured arterial pressure with Dubosq-Brazil solution after a brief washout with saline. After fixation, the renal tissue was weighed and 2 midcoronal sections were postfixed in buffered 4% formaldehyde solution. The material was then embedded in paraffin for assessment of glomerular and renal cortical interstitial injury and for immunohistochemical identification of lymphocytes and macrophages.

Histomorphometry
Sections 2–3 µm thick were stained with periodic acid-Schiff or Masson trichrome. All morphometric evaluations were performed in a blinded manner by a single observer.

Glomerular Injury
A detailed description of glomerular injury in the HS+N model is given elsewhere³ . The frequencies of glomerulosclerosis, glomerular ischemic collapse (COLL), and glomerular necrosis were evaluated by consecutive examination of at least 300 glomeruli at 400x.

Interstitial Injury
The fraction of the renal cortex occupied by interstitial tissue staining positively for extracellular matrix constituents was quantified in Masson-stained sections by a point-counting technique¹⁷ in 25 consecutive microscopic fields, at a final magnification of 100x under a 176-point grid.

Vascular Injury
The frequency of myointimal proliferation and fibrinoid necrosis in renal small arteries and arterioles was evaluated under 400x magnification in 3- to 4-µm-thick Masson-stained sections and expressed as a percentage of the total number of microvessels examined (55 per section in average). Myointimal fibrosis and microthrombosis were also noted in Group HS+N, but because their frequency was very small, these lesions were not included in the quantitative analysis.

Immunohistochemical Analysis
Macrophages and T-lymphocytes were detected in 4 µm-thick paraffin-embedded sections. These sections were mounted on glass slides coated with 2% gelatin, deparaffinized in xylene, and rehydrated through graded ethanol and then in deionized water. Sections were then microwave irradiated in citrate buffer to enhance antigen retrieval, and preincubated with 5% normal rabbit serum in Tris-buffered saline or in phosphate-buffered saline to prevent unspecific protein binding.

Optimal working dilutions of the primary antibodies were previously determined in titration experiments. Negative control experiments for all antigens were performed by omitting incubation with the primary antibody.

For specific immunostaining of T-lymphocytes, a monoclonal mouse anti-rat CD-3 antibody (Seralab, Oxford, UK) and an indirect streptavidin-biotin alkaline phosphatase technique were used. Sections were preincubated with normal horse serum to reduce nonspecific staining and then with avidin and biotin solutions to block nonspecific binding of these compounds. The incubation with the primary antibody was performed at room temperature for 90 minutes. Sections were then incubated at room temperature, first with rat-adsorbed biotinylated anti-mouse IgG (Vector Labs, Burlingame, Calif) for 45 minutes, and then with the streptavidin-biotin-alkaline phosphatase complex (Dako Co, Denmark) for 30 minutes.²⁰ Sections were finally incubated with a freshly prepared substrate, consisting of naphthol AS-MX-Phosphate and fast-red dye (Sigma Chemical Co), counterstained with Mayer’s hemalum and covered with Kaiser’s glycerin-gelatin (Merck).

For detection of macrophages, a monoclonal mouse anti-rat ED-1 antibody (Serotec, Oxford, UK) was used. Incubations were performed overnight at 4°C in a humidified chamber. After the sections were washed, they were incubated with rabbit anti-mouse immunoglobulin (Dako Co, Denmark). To complete the sandwich technique, incubation with a soluble complex of alkaline phosphatase anti-alkaline phosphatase (APAAP, Dako Co, Denmark) was performed. The last 2 steps were repeated to enhance the intensity of the reaction product. Finally, slides were developed with a fast-red dye solution and counterstained as in the lymphocyte detection procedure.

Quantitative analysis of ED-1- and CD-3-positive cells was performed in a blinded fashion by a single observer under 250x magnification. For each section, 25 microscopic fields, corresponding to a total area of 1.5 mm², were examined to assess the distribution of these cells among the glomerular (cells/tuft), microvascular (cells/transverse section), and tubulointerstitial (cells/mm²) compartments.

Statistics
One-way ANOVA with pairwise comparisons according to the Newmann-Kuls formulation was used in this study.¹⁷ P levels of 0.05 or less were considered significant.

	Results

Top Abstract Introduction Methods Results Discussion References

 
Renal function and hemodynamic parameters at 15 days of treatment are shown in Table 1. Body growth was similar among groups. Likewise, no differences in renal mass were observed at this stage. As described previously,³ blood pressure was markedly elevated in Group HS+ N. Hypertension was unchanged by MMF treatment. The HS+N treatment reduced glomerular filtration rate by 40%, an effect not reversed by MMF. Changes in renal plasma flow followed a similar pattern. P_GC was markedly elevated relative to HS in both Group HS+N and Group HS+N+MMF.

View this table: [in this window] [in a new window]   Table 1. Renal Functional and Hemodynamic Parameters at 15 Days of Treatment

Parameters obtained at 30 days after treatment are presented in Table 2. All groups gained weight compared with values measured at 15 days, although growth was blunted in Groups HS+N and HS+N+MMF. Tail-cuff pressure was elevated in the HS+N group. MMF treatment caused no significant attenuation of hypertension. U_albV was markedly increased in HS+N rats, reaching values 100-fold higher than the values of Group HS. Albuminuria was attenuated in Group HS+N+MMF, but remained elevated compared with HS. Likewise, P_creat was elevated in Group HS+N compared with HS. MMF treatment significantly reduced P_creat, which nevertheless remained high compared with HS.

View this table: [in this window] [in a new window]   Table 2. Body Weight, Tail-cuff Pressure, Albuminuria, and Plasma Creatinine at 30 Days of Treatment

Quantitative analysis of renal parenchymal and vascular injury at 30 days of treatment is given in Table 3. GS reached 3.5±0.7% in Group HS+N, compared with 0.4±0.2% in HS. MMF treatment reduced GS to levels indistinguishable from control. COLL represented almost one-third of all glomeruli in Group HS+N. MMF treatment reduced, but did not normalize, the frequency of COLL. Necrotic lesions appeared in 0.8±0.2% of glomeruli in Group HS+N. MMF treatment reduced the frequency of necrotic lesions to negligible values. The fraction of cortical parenchyma occupied by interstitial tissue was 0.5±0.1% in the HS group, increasing to 6.1±0.8% in the HS+N group, whereas MMF treatment reduced this proportion to 3.4±0.8 (P<0.05 versus HS+N and HS). Myointimal proliferation was noted in 9.4±1.8% of microvessels in Group HS+N and in 6.2±2% in Group HS+N+MMF (P>0.05). Fibrinoid necrosis appeared in 3.6±0.9% of microvessels in Group HS+N, as opposed to only 0.6±0.3% in Group HS+N+MMF (P<0.05).

View this table: [in this window] [in a new window]   Table 3. Quantification of Glomerular, Interstitial, and Vascular Injury at 30 Days of Treatment

Immunohistochemical analysis at 30 days of treatment is represented in Figure 1. The density of cells staining positively for the lymphocyte-specific CD-3 antigen was significantly increased compared with HS in glomeruli, microvessels, and interstitium. However, the vast majority of these infiltrating cells located at the interstitial area. MMF treatment significantly reduced cell infiltration in all compartments by nearly 50%. Likewise, a pronounced macrophage infiltration was observed in HS+N rats. The predominant location of ED-1-positive cells was again the interstitial area, with only a minority locating at the renal microvessels. No abnormal macrophage infiltration was observed at the glomeruli: the number of macrophages per glomerulus was similar between Groups HS+N and HS. MMF treatment reduced macrophage infiltration by 40% in both the interstitium and the microvessels. However, no effect was seen at the glomeruli.

View larger version (32K): [in this window] [in a new window]   Figure 1. Graphic representation of the extent of infiltrating cells staining positively for CD-3 (lymphocyte-specific, left panels) and ED-1 (macrophage-specific, right panels) antigens and their distribution among glomeruli, microvessels, and interstitium. *P<0.05 vs HS; #P<0.05 vs HS+N.

	Discussion

Top Abstract Introduction Methods Results Discussion References

 
As shown in previous studies by this laboratory,^{3 4} the association of chronic L-NAME treatment and salt overload resulted in systemic hypertension, creatinine retention, and severe renal injury characterized by massive albuminuria, COLL, glomerulosclerosis, glomerular necrosis, interstitial inflammation, and microvascular injury. The marked glomerular hypertension observed in HS+N rats likely contributed to the development of renal injury, as observed in other models.²¹ Additional damage may have derived from inflammatory phenomena, as suggested by the finding of marked renal lymphocyte and macrophage infiltration in HS+N rats, especially at interstitial areas. Renal inflammation may have resulted from glomerular hemodynamic stress, which may enhance the production of cytokines and growth factors^{6 22} and promote podocyte damage with formation of tuft synechiae²³ and leakage of glomerular filtrate into the interstitium.²⁴ Additional transmission of glomerular damage to the interstitium may have resulted from the massive proteinuria observed in HS+N rats, which can cause proximal tubular cells to produce inflammatory mediators as a result of their intense protein-absorptive activity.²⁵ Renal inflammation may also have resulted directly from NO inhibition.^{13 14} This effect is not unexpected, because NO is known to function as an endogenous anti-inflammatory agent by inhibiting cell proliferation,²⁶ leukocyte adhesion,⁸ the expression of adhesion molecules,²⁷ and platelet adhesion and activation.²⁸ Accordingly, chronic NO inhibition is associated with enhanced leukocyte-endothelium interaction⁹ and increased expression of ICAM-1/VCAM-1.¹⁰ Increasing evidence suggests that chronic inflammation, once regarded as an inseparable element of immune-mediated damage, plays a central role in the pathogenesis of progressive nephropathies of nonimmune origin as well. We¹⁷ and others^{6 18} showed intense renal interstitial lymphocyte infiltration in the 5/6 renal ablation model 1 week after surgery. Accordingly, we have recently shown that treatment of these rats with a nonsteroidal anti-inflammatory agent strongly attenuates renal injury.¹⁹ Even the beneficial effect of suppressors of the renin-angiotensin system in progressive renal diseases, usually attributed to its renal and glomerular hemodynamic effects, may have an anti-inflammatory component. Angiotensin II may enhance inflammation by promoting leukocyte infiltration, increasing the expression of adhesion molecules, and stimulating the cell-mediated immune response through activation of calcineurin phosphatase.^{29 30} Accordingly, recent studies have shown that tacrolimus, a calcineurin inhibitor, limits glomerular injury in the renal ablation model.³¹ In the chronic NO inhibition model, ACE inhibition reduced the expression of vascular adhesion molecules,¹⁰ whereas administration of an angiotensin II receptor blocker reduced cell proliferation, macrophage infiltration, and the expression of adhesion molecules.^{5 13}

Treatment of HS+N rats with MMF lowered albuminuria by >50%, reduced the frequency of all modalities of glomerular injury (with the exception of macrophage infiltration), and limited the extent of vascular and interstitial injury. Accordingly, serum creatinine was significantly reduced in MMF-treated rats, indicating relative renal functional preservation. These beneficial effects cannot be explained by amelioration of the renal or systemic hemodynamics because MMF did not change systemic or glomerular blood pressure compared with untreated HS+N rats. Renal protection was associated with attenuation of 2 major inflammation markers, namely renal lymphocyte and macrophage infiltration, particularly at interstitial areas. MMF has primarily been used in recent years as an immunosuppressor in the treatment of typically immune-mediated processes such as allograft rejection.^{15 16} In addition, recent experimental^{17 18 32 33} and clinical³⁴ evidence suggests that MMF therapy may retard the progression of chronic nephropathies of nonimmunologic nature alone^{17 18} or in combination with suppressors of the renin-angiotensin system.^{32 33} The basic mechanism thought to mediate these beneficial effects is a selective inhibition of lymphocyte proliferation.³⁵ Interstitial lymphocyte infiltration was strongly attenuated in MMF-treated HS+N rats, as previously reported in the renal ablation model.¹⁷ This effect may have limited inflammation by disrupting the interaction between lymphocytes and macrophages, thus reducing macrophage infiltration of the renal interstitium. These results underline the importance of cell-mediated immunity in the development of renal injury in this model. We have shown preliminary evidence of enhanced proliferation of glomerular, tubular, and interstitial cells in the chronic NO inhibition model.¹⁴ Thus MMF may have conferred additional protection by inhibiting the proliferation of mesangial³⁶ and tubular¹⁷ cells and by reducing the expression of adhesion molecules.¹⁸ Additional salutary effects may have derived from reduction of proteinuria.

In summary, MMF treatment attenuated renal injury in rats subjected to chronic NO inhibition by a nonhemodynamic mechanism, most likely associated with its anti-inflammatory action. Renal inflammation may be crucial to the development of renal injury in this model.

	Acknowledgments

 
This work was supported by grants No. 95/4710-2 and 98/09569-4 from the São Paulo Foundation for Research Support (FAPESP). During these studies, Roberto Zatz was the recipient of a Research Award (No. 326.429/81) from the Brazilian Council of Scientific and Technologic Development (CNPq).

Portions of this study were presented at the 31st Congress of the American Society of Nephrology, Philadelphia, Pennsylvania, USA - October, 1998, and published in abstract form (J Am Soc Nephrol. 1998;9:609A).

We are grateful to Cláudia R. Sena, Gláucia R. Antunes, Ivone Braga de Oliveira, Flávia R. Siqueira, and Marinete M. dos Santos for expert technical assistance.

Received February 18, 2000; first decision March 14, 2000; accepted June 30, 2000.

	References

Top Abstract Introduction Methods Results Discussion References

 
1. Ribeiro MO, Antunes E, De Nucci G, Lovisolo SM, Zatz R. Chronic inhibition of nitric oxide synthesis. A new model of arterial hypertension. Hypertension. 1992;20:298–303.[Abstract/Free Full Text]

2. Baylis C, Mitruka B, Deng A. Chronic blockade of nitric oxide synthesis in the rat produces systemic hypertension and glomerular damage. J Clin Invest. 1992;90:278–281.

3. Fujihara CK, Michellazzo SM, De Nucci G, Zatz R. Sodium excess aggravates hypertension and renal parenchymal injury in rats with chronic NO inhibition. Am J Physiol. 1994;266:F697–F705.[Abstract/Free Full Text]

4. Yamada SS, Sassaki AL, Fujihara CK, Malheiros DM, De Nucci G, Zatz R. Effect of salt intake and inhibitor dose on arterial hypertension and renal injury induced by chronic nitric oxide blockade. Hypertension. 1996;27:1165–1172.[Abstract/Free Full Text]

5. Casellas D, Benahmed S, Artuso A, Jover B. Candesartan and progression of preglomerular lesions in N(G)-nitro-L- arginine methyl ester hypertensive rats. J Am Soc Nephrol. 1999;10 Suppl 11:S230–S233.

6. Kliem V, Johnson RJ, Alpers CE, Yoshimura A, Couser WG, Koch KM, Floege J. Mechanisms involved in the pathogenesis of tubulointerstitial fibrosis in 5/6-nephrectomized rats. Kidney Int. 1996;49:666–678.[Medline] [Order article via Infotrieve]

7. Sugimoto H, Shikata K, Hirata K, Akiyama K, Matsuda M, Kushiro M, Shikata Y, Miyatake N, Miyasaka M, Makino H. Increased expression of intercellular adhesion molecule-1 (ICAM-1) in diabetic rat glomeruli: glomerular hyperfiltration is a potential mechanism of ICAM-1 upregulation. Diabetes. 1997;46:2075–2081.[Abstract]

8. Kubes P, Suzuki M, Granger DN. Nitric oxide: an endogenous modulator of leukocyte adhesion. Proc Natl Acad Sci U S A. 1991;88:4651–4655.[Abstract/Free Full Text]

9. Mitchell DJ, Yu J, Tyml K. Local L-NAME decreases blood flow and increases leukocyte adhesion via CD18. Am J Physiol. 1998;274:H1264–H1268.

10. Luvara G, Pueyo ME, Philippe M, Mandet C, Savoie F, Henrion D, Michel JB. Chronic blockade of NO synthase activity induces a proinflammatory phenotype in the arterial wall: prevention by angiotensin II antagonism. Arterioscler Thromb Vasc Biol. 1998;18:1408–1416.[Abstract/Free Full Text]

11. Tharaux PL, Chatziantoniou C, Casellas D, Fouassier L, Ardaillou R, Dussaule JC. Vascular endothelin-1 gene expression and synthesis and effect on renal type I collagen synthesis and nephroangiosclerosis during nitric oxide synthase inhibition in rats. Circulation. 1999;99:2185–2191.[Abstract/Free Full Text]

12. Chatziantoniou C, Boffa JJ, Ardaillou R, Dussaule JC. Nitric oxide inhibition induces early activation of type I collagen gene in renal resistance vessels and glomeruli in transgenic mice: role of endothelin. J Clin Invest. 1998;101:2780–2789.[Medline] [Order article via Infotrieve]

13. Bouriquet N, Dupont M, Herizi A, Mimran A, Casellas D. Preglomerular sudanophilia in L-NAME hypertensive rats: involvement of endothelin. Hypertension. 1996;27:382–391.[Abstract/Free Full Text]

14. Oliveira SG, Oliveira IB, Fujihara CK, Malheiros DMAC, De Nucci G, Zatz R, Noronha IL. Cellular events associated with renal injury in the chronic nitric oxide inhibition model. J Am Soc Nephrol. 1997;8:626A. Abstract.

15. Azuma H, Binder J, Heemann U, Schmid C, Tullius SG, Tilney NL. Effects of RS61443 on functional and morphological changes in chronically rejecting rat kidney allografts. Transplantation. 1995;59:460–466.[Medline] [Order article via Infotrieve]

16. Sollinger HW. Mycophenolate mofetil for the prevention of acute rejection in primary cadaveric renal allograft recipients. U.S. Renal Transplant Mycophenolate Mofetil Study Group. Transplantation. 1995;60:225–232.[Medline] [Order article via Infotrieve]

17. Fujihara CK, Malheiros DM, Zatz R, Noronha ID. Mycophenolate mofetil attenuates renal injury in the rat remnant kidney. Kidney Int. 1998;54:1510–1519.[Medline] [Order article via Infotrieve]

18. Romero F, Rodriguez-Iturbe B, Parra G, Gonzalez L, Herrera-Acosta J, Tapia E. Mycophenolate mofetil prevents the progressive renal failure induced by 5/6 renal ablation in rats. Kidney Int. 1999;55:945–955.[Medline] [Order article via Infotrieve]

19. Fujihara CK, Malheiros DM, Donato JL, Poli A, De Nucci G, Zatz R. Nitroflurbiprofen, a new nonsteroidal anti-inflammatory, ameliorates structural injury in the remnant kidney. Am J Physiol. 1998;274:F573–F579.

20. Noronha IL, Eberlein-Gonska M, Hartley B, Stephens S, Cameron JS, Waldherr R. In situ expression of tumor necrosis factor-alpha, interferon-gamma, and interleukin-2 receptors in renal allograft biopsies. Transplantation. 1992;54:1017–1024.[Medline] [Order article via Infotrieve]

21. Anderson S, Meyer TW, Rennke HG, Brenner BM. Control of glomerular hypertension limits glomerular injury in rats with reduced renal mass. J Clin Invest. 1985;76:612–619.

22. Lee LK, Meyer TW, Pollock AS, Lovett DH. Endothelial cell injury initiates glomerular sclerosis in the rat remnant kidney. J Clin Invest. 1995;96:953–964.

23. Nagata M, Kriz W. Glomerular damage after uninephrectomy in young rats. II. Mechanical stress on podocytes as a pathway to sclerosis. Kidney Int. 1992;42:148–160.[Medline] [Order article via Infotrieve]

24. Kriz W, Hosser H, Hahnel B, Gretz N, Provoost AP. From segmental glomerulosclerosis to total nephron degeneration and interstitial fibrosis: a histopathological study in rat models and human glomerulopathies. Nephrol Dial Transplant. 1998;13:2781–2798.[Abstract/Free Full Text]

25. Wang Y, Chen J, Chen L, Tay YC, Rangan GK, Harris DC. Induction of monocyte chemoattractant protein-1 in proximal tubule cells by urinary protein. J Am Soc Nephrol. 1997;8:1537–1545.[Abstract]

26. Scott-Burden T, Schini VB, Elizondo E, Junquero DC, Vanhoutte PM. Platelet-derived growth factor suppresses and fibroblast growth factor enhances cytokine-induced production of nitric oxide by cultured smooth muscle cells. Effects on cell proliferation. Circ Res. 1992;71:1088–1100.[Abstract/Free Full Text]

27. De Caterina R, Libby P, Peng HB, Thannickal VJ, Rajavashisth TB, Gimbrone MAJ, Shin WS, Liao JK. Nitric oxide decreases cytokine-induced endothelial activation. Nitric oxide selectively reduces endothelial expression of adhesion molecules and proinflammatory cytokines. J Clin Invest. 1995;96:60–68.

28. Radomski MW, Palmer RM, Moncada S. Endogenous nitric oxide inhibits human platelet adhesion to vascular endothelium. Lancet. 1987;2:1057–1058.[Medline] [Order article via Infotrieve]

29. Nataraj C, Oliverio MI, Mannon RB, Mannon PJ, Audoly LP, Amuchastegui CS, Ruiz P, Smithies O, Coffman TM. Angiotensin II regulates cellular immune responses through a calcineurin-dependent pathway. J Clin Invest. 1999;104:1693–1701.[Medline] [Order article via Infotrieve]

30. Mervaala EM, Muller DN, Park JK, Schmidt F, Lohn M, Breu V, Dragun D, Ganten D, Haller H, Luft FC. Monocyte infiltration and adhesion molecules in a rat model of high human renin hypertension. Hypertension. 1999;33:389–395.[Abstract/Free Full Text]

31. Hamar P, Peti-Peterdi J, Razga Z, Kovacs G, Heemann U, Rosivall L. Coinhibition of immune and renin-angiotensin systems reduces the pace of glomerulosclerosis in the rat remnant kidney. J Am Soc Nephrol. 1999;10 Suppl 11:S234–S238.

32. Fujihara CK, Noronha IL, Malheiros DMAC, Antunes GR, Oliveira IB, Zatz R. Combined mycophenolate mofetil and losartan therapy arrests established injury in the remnant kidney. J Am Soc Nephrol. 2000;11:283–290.[Abstract/Free Full Text]

33. Remuzzi G, Zoja C, Gagliardini E, Corna D, Abbate M, Benigni A. Combining an antiproteinuric approach with mycophenolate mofetil fully suppresses progressive nephropathy of experimental animals. J Am Soc Nephrol. 1999;10:1542–1549.[Abstract/Free Full Text]

34. Briggs WA, Choi MJ, Scheel PJJ. Successful mycophenolate mofetil treatment of glomerular disease. Am J Kidney Dis. 1998;31:213–217.[Medline] [Order article via Infotrieve]

35. Eugui EM, Mirkovich A, Allison AC. Lymphocyte-selective antiproliferative and immunosuppressive effects of mycophenolic acid in mice. Scand J Immunol. 1991;33:175–183.[Medline] [Order article via Infotrieve]

36. Hauser IA, Renders L, Radeke HH, Sterzel RB, Goppelt-Struebe M. Mycophenolate mofetil inhibits rat and human mesangial cell proliferation by guanosine depletion. Nephrol Dial Transplant. 1999;14:58–63. [Abstract/Free Full Text]

This article has been cited by other articles:

				E. Tapia, D. J. Sanchez-Gonzalez, O. N. Medina-Campos, V. Soto, C. Avila-Casado, C. M. Martinez-Martinez, R. J. Johnson, B. Rodriguez-Iturbe, J. Pedraza-Chaverri, M. Franco, et al. Treatment with pyrrolidine dithiocarbamate improves proteinuria, oxidative stress, and glomerular hypertension in overload proteinuria Am J Physiol Renal Physiol, November 1, 2008; 295(5): F1431 - F1439. [Abstract] [Full Text] [PDF]

				S. D. Crowley, C. W. Frey, S. K. Gould, R. Griffiths, P. Ruiz, J. L. Burchette, D. N. Howell, N. Makhanova, M. Yan, H.-S. Kim, et al. Stimulation of lymphocyte responses by angiotensin II promotes kidney injury in hypertension Am J Physiol Renal Physiol, August 1, 2008; 295(2): F515 - F524. [Abstract] [Full Text] [PDF]

				M. L. Graciano, A. Nishiyama, K. Jackson, D. M. Seth, R. M. Ortiz, M. C. Prieto-Carrasquero, H. Kobori, and L. G. Navar Purinergic receptors contribute to early mesangial cell transformation and renal vessel hypertrophy during angiotensin II-induced hypertension Am J Physiol Renal Physiol, January 1, 2008; 294(1): F161 - F169. [Abstract] [Full Text] [PDF]

				M. L. Graciano, C. R. Mouton, M. E. Patterson, D. M. Seth, J. J. Mullins, and K. D. Mitchell Renal vascular and tubulointerstitial inflammation and proliferation in Cyp1a1-Ren2 transgenic rats with inducible ANG II-dependent malignant hypertension Am J Physiol Renal Physiol, June 1, 2007; 292(6): F1858 - F1866. [Abstract] [Full Text] [PDF]

				C. K. Fujihara, C. R. Sena, D. M. A. C. Malheiros, A. L. Mattar, and R. Zatz Short-term nitric oxide inhibition induces progressive nephropathy after regression of initial renal injury Am J Physiol Renal Physiol, March 1, 2006; 290(3): F632 - F640. [Abstract] [Full Text] [PDF]

				J. Mauro Vieira Jr, E. Mantovani, L. Tavares Rodrigues, H. Delle, I. L. Noronha, C. K. Fujihara, and R. Zatz Simvastatin attenuates renal inflammation, tubular transdifferentiation and interstitial fibrosis in rats with unilateral ureteral obstruction Nephrol. Dial. Transplant., August 1, 2005; 20(8): 1582 - 1591. [Abstract] [Full Text] [PDF]

				M. L. Graciano, R. d. C. Cavaglieri, H. Delle, W. V. Dominguez, D. E. Casarini, D. M. A. C. Malheiros, and I. L. Noronha Intrarenal Renin-Angiotensin System Is Upregulated in Experimental Model of Progressive Renal Disease Induced by Chronic Inhibition of Nitric Oxide Synthesis J. Am. Soc. Nephrol., July 1, 2004; 15(7): 1805 - 1815. [Abstract] [Full Text] [PDF]

				R. C. Campbell The Renin-Angiotensin System: A 21st Century Perspective J. Am. Soc. Nephrol., July 1, 2004; 15(7): 1963 - 1964. [Full Text] [PDF]

				R. Zatz, I. L. Noronha, and C. K. Fujihara Experimental and clinical rationale for use of MMF in nontransplant progressive nephropathies Am J Physiol Renal Physiol, December 1, 2002; 283(6): F1167 - F1175. [Abstract] [Full Text] [PDF]

				M. Y. Karim, P. Alba, M.-J. Cuadrado, I. C. Abbs, D. P. D'Cruz, M. A. Khamashta, and G. R. V. Hughes Mycophenolate mofetil for systemic lupus erythematosus refractory to other immunosuppressive agents Rheumatology, August 1, 2002; 41(8): 876 - 882. [Abstract] [Full Text] [PDF]

				I. L. Noronha, C. K. Fujihara, and R. Zatz The inflammatory component in progressive renal disease--are interventions possible? Nephrol. Dial. Transplant., March 1, 2002; 17(3): 363 - 368. [Full Text] [PDF]

This Article Abstract Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Fujihara, C. K. Articles by Zatz, R. Search for Related Content PubMed PubMed Citation Articles by Fujihara, C. K. Articles by Zatz, R. Related Collections Animal models of human disease Hypertension - basic studies Endothelium/vascular type/nitric oxide