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Int J Clin Exp Med 2018;11(3):1726-1733www.ijcem.com /ISSN:1940-5901/IJCEM0058003

Original ArticleIrbesartan inhibits the formation of calcium oxalate stones in the kidney of diabetic rats

Lei-Hua Cao1, Qi Chen2, Jia Chen3, Wei-Min Chen2

1Department of Urology, The Third Hospital of Nanchang, Nanchang 330003, China; 2Department of Urology, The First Affiliated Hospital of Nanchang University, Nanchang 330006, China; 3Department of Urology, Shaoxing Municipal Hospital, Shaoxing 312000, China

Received October 21, 2016; Accepted November 14, 2017; Epub March 15, 2018; Published March 30, 2018

Abstract: Objective: To investigate the composition and formation mechanism of kidney stones and effect of irbesar-tan on the formation of nephrolithiasis in diabetic rats. Methods: Totally speaking, 60 Sprague & Dawley (SD) rats were selected (8 weeks-old, 240-280 g) and randomly divided into four groups as followings: control group, control-Irbesartan group, diabetic group, and diabetic-Irbesartan group. To each group, the oxalic acid was measured via potassium chromate oxidation of methyl red by catalytic spectrophotometry; the concentration of calcium ion (Ca2+) and uric acid were measured by the automatic biochemical analyzer 6 months later. Meanwhile, the physiologi-cal levels of angiotensin II and hyaluronic acid (HA), the expression of renal osteopontin (OPN), the pathological changes and the calcium oxalate crystals were detected under the help of ELISA assays, western blotting, HE stain-ing, and environmental scanning electron microscopy one after another in kidney tissue. Results: As a result, the concentration of 24-hour uric acid in diabetic rats is significantly lower than that of normal rats (P < 0.05) and that in diabetic-Irbesartan rats is markedly lower than that of control-Irbesartan rats (P < 0.05). But to urine oxalate and calcium ion, compared with control-Irbesartan rats, the higher concentration can be detected in diabetic-Irbesartan rats (P < 0.05). Furthermore, the levels of hyaluronic acid and angiotensin II, the expression of OPN in the kidneys of diabetic rats are decreased by Irbesartan. At the same time, irbesartan ameliorated the abnormal pathological changes and hyaline degeneration of renal tubular epithelial cells in diabetic rats. Finally, the Irbesartan has an inhibitory effect on the formation of calcium oxalate crystals in diabetic rats. Conclusions: Irbesartan can protect diabetic kidney from abnormal pathological changes and calcium oxalate stones formation, which is related to pre-venting OPN expression.

Keywords: Irbesartan, diabetic, angiotensin II receptor antagonist, calcium oxalate kidney stones

Introduction

Urolithiasis is a growing health problem in industrialized countries and often correlated with habits, such as hypertension, high purine intake, diabetes, obesity and metabolic syn-drome. Unexpectedly, 12% of the world’s popu-lation is affected by urinary system stone dis-ease [1]. Urolithiasis is the formation of kidney stones in urine-collecting spaces of the kid-neys. Under some certain conditions, sub-stances normally dissolved in the urine can separate out as crystals and accumulate to form a solid mass called kidney stone. Stones could migrate into the ureters, the bladder and finally be evacuated in the urine [2].

With the rapid economic development and improvement of living standards, the preva-lence of diabetes is rapidly increasing. Diabetes complicated with urinary tract stones in patients was common in clinical practice. Meydan N found that the incidence of urinary tract stones in diabetes was higher than that of urinary calculi incidence of ordinary people. It indicated that diabetes and urinary tract stones could have a certain relationship [3]. Lieske JC found that diabetes is a predisposing factor for kidney stones. They believed that there was a significant correlation in diabetes and kidney stones and further proved that the ratio of dia-betic patients with uric acid stone formation is much higher than non-diabetic patients and

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Irbesartan inhibits the formation of calcium oxalate kidney stones

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diabetes was an independent risk factor for the formation of uric acid stones [4]. Daudon M showed that diabetes mellitus caused by the ability to damage the secretion of ammonia so as to reduce the pH value of urine. The reduc-tion of urinary PH value is the main factor in the formation of uric acid stones, so diabetic patients are more likely to form uric acid stones [5].

The angiotensin-II-receptor antagonist Irbesar- tan was associated with better renal outcomes than the other agents (amlodipine, placebo, and antihypertensive agents) we used. In recent years, a large number of studies had shown that Irbesartan has the role of anti-dia-betic renal fibrosis. Irbesartan can not only reduce the osteopontin (OPN) and HA expres-sion in the kidney content [6], but also protect the renal cells by reducing the damage of the reactive oxygen species in the glomerular endo-thelial cells by reducing the content of Ang II in the kidney [7]. It can be seen that Irbesartan not only have the effect of protecting renal cell damage, but also has the effect of decreasing the expression of OPN and HA, which are not conducive to the formation of stones.

In this study, we established diabetic animal model to uncover the difference in the compo-sition of the kindey stones compared with nor-mal control, and the underlying mechanism. Then we further researched the act of Ir- besartan on the formation of diabetic nephroli-thiasis under hyperglycemia.

Materials and methods

Animals and ethics

Sixty male Sqrague-Dawley rats (240-280 g) were obtained from the Laboratory Animal Centre of Medical College of Nanchang University (Jiangxi, China). All animal proce-dures were approved and conducted in accor-dance with the guidelines for the care and use of animal of the ethics committee of Nanchang University.

General reagents and equipment

All the regents and assay kits were from com-mercial sources. Streptozotocin (STZ) was pur-chased from Solarbio Life Science (Beijing, China). Irbesartan was purchased from Haiyan

Pharmaceutical Co., Ltd (Beijing, China). Both rat hyaluronic acid (HA) ELISA kit and angioten-sin II (ANG-II) ELISA kit came from Xinyu Biotech Co., Ltd (Shanghai, China). BCA Protein Assay Kit was purchased from Tiangen Biotech Co., Ltd (Beijing, China). Microscope (OLYMPUS, Germany) was used for section observation.

Grouping and surgical procedures

The rats were randomly divided into four groups (15 rats each): control group (normal rats), con-trol-Irbesartan group (normal rats treated with 50 mg/kg Irbesartan), diabetic group (diabetic rats), and diabetic-Irbesartan group (diabetic rats treated with 50 mg/kg Irbesartan).

Establishment of diabetic rats

SD rats were fed with high fat diet for 2 months before STZ injection, and then intraperitoneally injected with a single dose of STZ (50 mg/kg body weight) that dissolved in 0.01 M citrate buffer immediately before used [8]. After injec-tion, animals had free access to food and water. Tail-vein blood glucose was detected on the third day after STZ injection. SD rats with blood glucose values > 16.7 mmol/L indicated suc-cessful diabetic model.

Urine sample and kidney tissues collection

Rats in control group and diabetic group were orally administrated with 0.2% carboxy methyl cellulose sodium (CMC-Na) as a vehicle every-day. Meanwhile, rats in control-Irbesartan group and diabetic-Irbesartan group were orally administrated with 50 mg/kg Irbesartan (dis-solved in 0.2% CMC-Na) everyday. After 6 months of treatment that mention above, the animals were fasted overnight. Then 24 h urine samples were collected and centrifuged at 3000 rpm for 10 min, the supernatants were stored at -80°C for the measurement of oxalic acid, calcium ion (Ca2+), and uric acid. The oxal-ic acid of in each group was measured via potassium chromate oxidation of methyl red by catalytic spectrophotometry. Calcium ion (Ca2+) and uric acid were measured by the automatic biochemical analyzer.

After urine collection, all rats were killed under ether anesthesia. The left kidneys were imme-diately removed and rinsed several times in ice-saline, then stored at -80°C for subsequent


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detection. The right kidneys were cut into blocks and fixed by 10% neutral formalin for

0.05 was considered to be statistically significant.

Figure 1. The results of urinary oxalate, calcium ion and uric acid in 24-hour urine of SD rats. A: normal rats, B: normal rats treated with 50 mg/kg Irbe-sartan, C: diabetic rats, D: diabetic rats treated with 50 mg/kg Irbesartan.*p < 0.05, compared with A; #p < 0.05, compared with B.

slice observation. The protein expression levels of osteo-pontin (OPN) of the kidney tis-sues were quantified by west-ern blot. The HA and ANG-II were measured by enzyme-linked immunosorbent assays (ELISA) according to the instruction of the kits. The pathological changes and his-tologic evaluation were observed by microscope after hematoxylin and eosin (HE) staining and Von Kossa stain-ing, respectively (GENMED, Shanghai, China). Calcium oxalate crystals accumulation in kidney cells were observed by environmental scanning electron microscopy (Quanta 200F, FEI, America).

Western blot analysis

Total proteins were separated by 12% SDS-PAGE and then transferred to polyvinylidene difluoride membranes (Immo- bilon-P, Bio-Rad, America). Blots were incubated with OPN antibody (1:200, Bioss, China) and thereafter with HRP-conjugated goat anti-ra- bbit antibody (1:4000, Boster, China). Immunoreactive pro-tein was detected using the enhanced chemiluminescen ce method according to the manufacturer (Amersham, Ay- lesbury, UK).

Statistical analysis

Results were derived from three independent experi-ments and analyzed with SPSS 17.0 software (SPSS, Chicago, USA). Data was shown as mean ± SD. Sta- tistical analyses were per-formed using one-way analy-sis of variance. The P value <

Figure 2. The results of hyaluronic acid and angiotensin II in renal tissue of rats. A. The results of hyaluronic acid in renal tissue; B. The results of angiotensin II in renal tissue. A: normal rats, B: normal rats treated with 50 mg/kg Irbesartan, C: diabetic rats, D: diabetic rats treated with 50 mg/kg Irbesartan. *p < 0.05.


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Results

Uric acid, oxalate and calcium ions in the urine of diabetic rats with high blood glucose and normal rats were not affected by Irbesartan

As shown in Figure 1, the 24-hour urine uric acid levels in diabetic rats and diabetic-Irbesar-tan rats were significantly lower than that of normal rats and normal-Irbesartan rats respectively (P < 0.05). On the other hand, the 24-hour urine oxalate and calcium ion concen-

percentage of gray bands were calculated and analyzed by LSD-t test. As shown in Figure 3A and 3B, compared with normal kidney, the expression of osteopontin in diabetic kidney was increased obviously. However, Irbesartan can decrease the expression of osteopontin in both normal and diabetic kidney interestingly. As a result, we can conclude that OPN expres-sion in diabetic kidney was much higher than that of normal kidney and Irbesartan fights against the OPN expression in both normal and diabetic rats.

Figure 3. The results of western blot analysis of OPN protein in kidney. A. The expression level of OPN protein; B. The percentage of grayscale values for the stripe and the internal reference band. A: normal rats, B: normal rats treated with 50 mg/kg Irbesartan, C: diabetic rats, D: diabetic rats treated with 50 mg/kg Irbesartan. *p < 0.05.

tration of diabetic rats were significantly higher than that in normal rats (P < 0.05). However, there was no signifi-cant difference between dia-betic rats and diabetic-Irbe-sartan rats in urine uric acid levels, urine oxalate, and cal-cium ion concentration (P > 0.05).

Irbesartan decreased levels of hyaluronic acid and angio-tensin II in renal tissues of rats

The levels of hyaluronic acid and angiotensin II in renal tis-sues of SD rats were deter-mined. As shown in Figure 2A and 2B, angiotensin II and hyaluronic acid content in renal tissue of diabetic rats were significantly higher than that in normal rats (P < 0.05). Irbesartan could significantly inhibit the levels of hyaluronic acid and angiotensin II, indi-cating that Irbesartan played an important role in lowering the risk factors of kidney-stone formation in hyperglyce-mia and diabetes.

Irbesartan decreased OPN protein in renal tissues of rats

The content of OPN and β-actin in the kidneys were analyzed quantitatively by ImageJ 1.48u software. The

Figure 4. The HE staining of paraffin sections of renal (x400). A. Normal rats, B. Normal rats treated with 50 mg/kg Irbesartan, C. Diabetic rats, D. Dia- betic rats treated with 50 mg/kg Irbesartan. Arrows: epithelial damage (swelling, degeneration, necrosis, tubular expansion, or hyaline degenera-tion).


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Irbesartan prevented abnormal pathological changes in kidney

Pathological changes of renal paraffin sections were observed by HE staining. From the Figure 4A and 4B, we could see that there were no abnormalities in both normal group and nor-mal-Irbesartan group. However, renal tubular epithelial cells of diabetic rats were oncotic, degenerative, or even necrotic with hyaline degeneration, which is the typical characteris-tic of abnormal pathological changes (Figure 4C and 4D). The abnormal pathological chang-es of renal tubular epithelial cells were amelio-rated by Irbesartan-treatment, indicating that Irbesartan was able to prevent abnormal path-ological changes in the diabetic kidney (Figure 4D).

rats with high blood glucose was decreased and lower than that in normal rats. The causes may be as follows: first, elevated blood glucose level in diabetes rats causes increased insulin resistance [9], which could reduce uric acid clearance, leading to decreased urinary uric acid [10]. Second, sharply and frequently fluctu-ated blood glucose level of day and night was more prone to damage the renal tubular cells [11]. With high blood glucose, accelerated renal dysfunction could lead to decreasing renal tubular excretion of uric acid, leading to decreased urinary uric acid. It’s well known that the reduction of urinary uric acid was not conducive to the formation of uric acid stones. So diabetes with high blood glucose was not conducive to the formation of uric acid stones.

Figure 5. The results of Von-Kossa staining of kidney tissues (x 400). A. Nor-mal rats, B. Normal rats treated with 50 mg/kg Irbesartan, C. Diabetic rats, D. Diabetic rats treated with 50 mg/kg Irbesartan. Arrows: calcium oxalate kidney stones.

Irbesartan inhibited calcium oxalate crystals formation in diabetic kidney

The buildup of calcium oxa-late crystals in Kidney tissue was observed by Von-Kossa staining and environmental scanning electron microsco-py. As shown in Figures 5 and 6, there were no calcium oxa-late crystals in the kidney tis-sues of normal rats but oppo-site consequences in kidney tissue of diabetic rats. After treated with Irbesartan, the amount of crystals was reduced in diabetic kidneys. Consequences above indicat-ed that Irbesartan can inhibit or delay the formation of cal-cium oxalate kidney stones in diabetic rats.

Discussion

24 hours Uric acid concentra-tion detection showed that urinary uric acid concentra-tion in diabetic rats was lower than that in normal rats. Also, uric acid concentration in normal rats treated with Ir- besartan was lower than that in diabetic rats treated with Irbesartan. So urinary uric acid concentration in diabetic


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24 hours urinary oxalate, calcium ion concen-trations detection showed that urinary oxalate and calcium ion concentrations in diabetic rats were higher than that of normal rats. On the other hand, urinary oxalate and calcium ion concentrations in normal rats treated with Irbesartan was higher than that in diabetic rats treated with Irbesartan. The reason for increased urinary oxalate in diabetes with high blood glucose may be as follows: lack of intesti-nal formic acid oxalic acid bacteria in diabetes reduced the decomposition of intestinal oxalic acid, leading to increased intestinal absorption of oxalate and increased urinary oxalate excre-tion [12]. Brian H Eisner and CT Lee have

exceeded their own free radical scavenging rate [17], leading to directly damaging the renal cells [18]. From the above, we could see that renal cells were damaged in diabetes with high blood glucose. Damaged renal cells were con-ducive to the formation of calcium oxalate kid-ney stones. So diabetes with high blood glu-cose was conducive to the formation of calcium oxalate kidney stones.

OPN is a phosphate protein with a relative molecular mass of 44 Kda [19]. Approximately half of the amino acids of the protein are aspar-tic acids and glutamic acids. OPN have multiple binding sites including “integrin sites” and “cal-

Figure 6. The environmental scanning electron microscopy picture of Kidney tissues (x6000). A. Normal rats, B. Normal rats treated with 50 mg/kg Irbe-sartan, C. Diabetic rats, D. Diabetic rats treated with 50 mg/kg Irbesartan. Arrows: calcium oxalate crystal.

reported that urinary oxalate and calcium concentrations in the urine were supersaturat-ed and were higher than those in normal urine [13, 14], which is consistent with our experi-ment results.

Renal Ang II examination showed that the renal Ang II content in diabetic rats was significantly higher than that in normal rats. On the other hand, Ang II in normal rats treated with Irbesartan was higher than that in diabetic rats treated with Irbesartan. AVidotti DB had put forward that in vivo Ang II under high blood glucose was increased in the study of angiotensin II metabolism mechanism of diabetic rats [15]. The conclu-sion was the same as our experiment result. As the in- crease in Ang II could cause renal cell damage by free radi-cals [16], our results indirectly indicated that renal cells were impaired in diabetes with high blood glucose. The renal par-affin section HE staining results showed that the renal cells in diabetic rats with high blood glucose were signifi-cantly impaired. Damaged renal cells may be caused by excessive free radicals under high blood glucose which has


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cium ion binding sites” [20, 21]. The integrin sites of OPN could be attached to cells by bind-ing to multiple integrin receptors on the cell sur-face [22]. The calcium binding sites of OPN were prone to bind calcium ions in calcium oxa-late. OPN is not only related to bone metabo-lism, cardiovascular system, and tumor, but also related to the urinary system. Free-state normal-structured OPN is an inhibitor of calci-um oxalate crystal growth and aggregation [23]. However, renal epithelial cells that can’t be attached with calcium oxalate crystals can be converted to renal epithelial cells to promote the formation of calcium oxalate stones [24]. Thus, OPN can promote the formation of calci-um oxalate kidney stones instead of inhibiting it. According to the results of HA content in the kidney, we knew that the HA content in the kid-ney of diabetic rats was higher than that of nor-mal rats. Meanwhile, HA content in normal rats treated with Irbesartan was higher than that in diabetic rats treated with Irbesartan. Study by Yevdokimova N Y has confirmed that in the high-glucose environment, renal synthesis of HA was elevated through increasing in renal HA content and prostaglandins [25]. HA cannot only moisturize, but also prevent arteriosclero-sis and cancer. Also, HA is closely related to the formation of urinary tract stones. A large num-ber of studies have shown that HA was one of the main adhesion molecules of calcium oxa-late crystals, promoting the formation of calci-um oxalate stones [26]. In the diabetic kidney under high blood glucose, OPN and HA con-tents promoted the formation of calcium oxa-late kidney stones.

We know that urinary oxalate and calcium ions in the urine are super-saturate in diabetes. At the same time, damaged kidney cells, high-expressed OPN and HA were risk factors to pro-mote the formation of calcium oxalate stones [27]. Therefore, calcium oxalate kidney stones are possibly formed in diabetes.

With von Kusa staining, calcium oxalate kidney stones in diabetes were observed [28]. We observed black oxalate crystal deposition in diabetic rats. Although von Kusa staining can detect calcium oxalate, it still has defects. Thus we used environmental electron microscopy to further confirm the e calcium oxalate crystal accumulation on renal cell surface. We found calcium oxalate crystal adhesion on diabetic

kidney cell surface. But Irbesartan could effec-tively prevent calcium oxalate crystal adhesion on renal cell surface, indicating that Irbesartan could inhibit or delay the formation of calcium oxalate kidney stones in diabetes.

According to the detection of related factors in the formation of calcium oxalate kidney stones and renal pathological changes, we found that uric acid, oxalate and calcium ions in the urine of diabetic rats with high blood glucose and normal rats were not affected by Irbesartan. But Irbesartan could reduce OPN, HA, and Ang II contents in normal rats and diabetic kidneys and weaken renal cell damage. In conclusion, Irbesartan inhibited or delayed calcium oxalate kidney stone formation in diabetes was related to reduced OPN and HA contents and weaken the renal cell oxidative damage, which finally prevented the aggregation and adhesion of cal-cium oxalate crystals.

Disclosure of conflict of interest

None.

Address correspondence to: Wei-Min Chen, Department of Urology, First Affiliated Hospital of Nanchang University, No. 17, Yongwai Street, Donghu District, Nanchang City 330006, Jiangxi Province, China. Tel: 13970010866; E-mail: [email protected]

References

[1] Rosa M, Usai P, Miano R, Kim FJ, Agrò EF, Bove P and Micali S; International Translational Re-search in Uro-Sciences Team (ITRUST). Recent finding and new technologies in nephrolithia-sis: a review of the recent literature. BMC Urol 2013; 13: 10.

[2] Orhan N, Onaran M, Sen I, Isik Gonul I and Aslan M. Preventive treatment of calcium oxa-late crystal deposition with immortal flowers. J Ethnopharmacol 2015; 163: 60-67.

[3] Meydan N, Barutca S, Caliskan S and Camsari T. Urinary stone disease in diabetes mellitus. Scand J Urol Nephrol 2003; 37: 64-70.

[4] Lieske JC, de la Vega LS, Gettman MT, Slezak JM, Bergstralh EJ, Melton LJ and Leibson CL. Diabetes mellitus and the risk of urinary tract stones: a population-based case-control study. Am J Kidney Dis 2006; 48: 897-904.

[5] Daudon M, Traxer O, Conort P, Lacour B and Jungers P. Type 2 diabetes increases the risk for uric acid stones. J Am Soc Nephrol 2006; 17: 2026-2033.

mailto:[email protected]


1733 Int J Clin Exp Med 2018;11(3):1726-1733

[6] Cao Z, Cox A and Bonnet F. Increased osteo-pontin expression following renal ablation is attenuated by angiotensin type 1 receptor an-tagonism. Exp Nephrol 2002; 10: 19-25.

[7] Pan Q, Yang XH and Cheng YX. Angiotensin II stimulates MCP-1 production in rat glomerular endothelial cells via NAD(P)H oxidase-depen-dent nuclear factor-kappa B signaling. Braz J Med Biol Res 2009; 42: 531-536.

[8] Punithavathi VR, Anuthama R and Prince PS. Combined treatment with naringin and vitamin C ameliorates streptozotocin-induced diabetes in male Wistar rats. J Appl Toxicol 2008; 28: 806-813.

[9] Haber CA, Lam TK, Yu Z, Gupta N, Goh T, Bog-danovic E, Giacca A and Fantus IG. N-acetyl-cysteine and taurine prevent hyperglycemia-induced insulin resistance in vivo: possible role of oxidative stress. Am J Physiol Endocrinol Metab 2003; 285: E744-753.

[10] So A and Thorens B. Uric acid transport and disease. J Clin Invest 2010; 120: 1791-1799.

[11] Monnier L, Mas E, Ginet C, Michel F, Villon L, Cristol JP and Colette C. Activation of oxidative stress by acute glucose fluctuations compared with sustained chronic hyperglycemia in pa-tients with type 2 diabetes. JAMA 2006; 295: 1681-1687.

[12] Kaufman DW, Kelly JP, Curhan GC, Anderson TE, Dretler SP, Preminger GM and Cave DR. Oxalobacter formigenes may reduce the risk of calcium oxalate kidney stones. J Am Soc Nephrol Jasn 2008; 19: 1197-1203.

[13] Lee CT, Lien YH, Lai LW, Chen JB, Lin CR and Chen HC. Increased renal calcium and magne-sium transporter abundance in streptozotocin-induced diabetes mellitus. Kidney Int 2006; 69: 1786-1791.

[14] Eisner BH, Porten SP, Bechis SK and Stoller ML. Diabetic kidney stone formers excrete more oxalate and have lower urine pH than nondiabetic stone formers. J Urology 2010; 183: 2244-2248.

[15] Vidotti DB, Casarini DE, Cristovam PC, Leite CA, Schor N and Boim MA. High glucose concen-tration stimulates intracellular renin activity and angiotensin II generation in rat mesangial cells. Am J Physiol Renal Physiol 2004; 286: F1039-1045.

[16] Ruiz-Ortega M, Ruperez M, Lorenzo O, Esteban V, Blanco J, Mezzano S and Egido J. Angioten-sin II regulates the synthesis of proinflamma-tory cytokines and chemokines in the kidney. Kidney Int 2002; 82: S12-22.

[17] Rahimi R, Nikfar S, Larijani B and Abdollahi M. A review on the role of antioxidants in the man-agement of diabetes and its complications. Biomed pharmacother 2005; 59: 365-373.

[18] Nishikawa T, Edelstein D, Du XL, Yamagishi S, Matsumura T, Kaneda Y, Yorek MA, Beebe D, Oates PJ and Hammes HP. Normalizing mito-chondrial superoxide production blocks three pathways of hyperglycaemic damage. Nature 2000; 404: 787-790.

[19] Kahles F, Findeisen HM and Bruemmer D. Os-teopontin: a novel regulator at the cross roads of inflammation, obesity and diabetes. Mol Metab 2014; 3: 384-393.

[20] Riese RJ, Mandel NS, Wiessner JH, Mandel GS, Becker CG and Kleinman JG. Cell polarity and calcium oxalate crystal adherence to cultured collecting duct cells. Am J Physiol 1992; 262: F177-184.

[21] Nishimichi N, Higashikawa F, Kinoh HH, Tatei-shi Y, Matsuda H and Yokosaki Y. Polymeric osteopontin employs integrin alpha9beta1 as a receptor and attracts neutrophils by present-ing a de novo binding site. J Biol Chem 2009; 284: 14769-14776.

[22] Liu L, Chen S, Giachelli CM, Ratner BD and Ji-ang S. Controlling osteopontin orientation on surfaces to modulate endothelial cell adhe-sion. J Biomed Mater Res A 2005; 74A: 23-31.

[23] O’Young J, Chirico S, Tarhuni NA, Grohe B, Kart-tunen M, Goldberg HA and Hunter GK. Phos-phorylation of osteopontin peptides mediates adsorption to and incorporation into calcium oxalate crystals. Cells Tissues Organs 2009; 189: 51-55.

[24] Asselman M, Verhulst A, De Broe ME and Verkoelen CF. Calcium oxalate crystal adher-ence to hyaluronan-, osteopontin-, and CD44-expressing injured/regenerating tubular epi-thelial cells in rat kidneys. J Am Soc Nephrol 2003; 14: 3155-3166.

[25] Yevdokimova NY. Elevated level of ambient glu-cose stimulates the synthesis of high-molecu-lar-weight hyaluronic acid by human mesangial cells. The involvement of transforming growth factor beta1 and its activation by thrombos-pondin-1. Acta Biochim Pol 2006; 53: 383-393.

[26] Lamontagne CA, Plante GE and Grandbois M. Characterization of hyaluronic acid interaction with calcium oxalate crystals: implication of crystals faces, pH and citrate. J Mol Recognit 2011; 24: 733-740.

[27] Khan SR. Renal tubular damage/dysfunction: key to the formation of kidney stones. Urolithia-sis 2006; 34: 86-91.

[28] Gáspár S, Niculiţe C, Cucu D and Marcu I. Ef-fect of calcium oxalate on renal cells as re-vealed by real-time measurement of extracel-lular oxidative burst. Biosens Bioelectron 2010; 25: 1729-1734.

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