Guanosine protects against glycerol-induced acute kidney injury via up-regulation of the klotho gene

Document Type : Original Article


1 Department of Medical Biochemistry, Faculty of Medicine, Minia University, 61511, Minia, Egypt

2 Department of Pharmacology, Faculty of Medicine, Minia University, 61511, Minia, Egypt

3 Department of Pathology, Faculty of Medicine, Minia University, 61511, Minia, Egypt


Objective(s): Acute Kidney Injury (AKI) is characterized by a rapid and reversible decline in renal function with a rapid decrease in Glomerular Filtration Rate (GFR), which is associated with high mortality. Rhabdomyolysis accounts for 10–40% of AKI, to which the therapeutic approach is limited. Klotho is a protein that modulates sodium-phosphate co-transporters, ion channels that have been reported to have a renal protective effect. Guanosine, a purine nucleoside, has already been reported to have a renal protective effect; however, the mechanism of such protection and its relation to Klotho modification has not been evaluated yet. This study aims to evaluate the mechanism of the protective effect of guanosine against rhabdomyolysis-induced AKI and its relation to the expression of the Klotho gene.
Materials and Methods: In the current study, rats were divided into three groups: control, glycerol-induced AKI, and guanosine-treated. Serum urea and creatinine levels, renal tissue Total Antioxidant Capacity (TAC), and Klotho and Cystatin C genes expression were evaluated. Furthermore, caspase-3 immunostaining and histopathological evaluations were done. 
Results: Results showed that guanosine treatment resulted in a significant reduction in serum urea and creatinine, Cystatin C genes expression, and caspase-3 immunoexpression, and an increase in TAC and Klotho genes expression. Results also revealed an improvement of renal histopathology when compared with the glycerol-induced AKI group. 
Conclusion: Guanosine may be a promising agent in the treatment of rhabdomyolysis-induced AKI. The proposed mechanism for guanosine may be through its ability to enhance Klotho gene expression in renal tissue, with subsequent antioxidant and anti-apoptotic activity.


1. Kim JH, Lee SS, Jung MH, Yeo HD, Kim H-J, Yang JI, et al. N-acetylcysteine attenuates glycerol-induced acute kidney injury by regulating MAPKs and Bcl-2 family proteins. Nephrol Dial Transplant 2010; 25:1435-1443.
2. Ward MM. Factors predictive of acute renal failure in rhabdomyolysis. Arch Intern Med 1988; 148:1553-1557.
3. Homsi E, Janino P, de Faria JB. Role of caspases on cell death, inflammation, and cell cycle in glycerol-induced acute renal failure. Kidney Int 2006; 69:1385-1392.
4. Elmore S. Apoptosis: a review of programmed cell death. Toxicologic pathology 2007; 35:495-516.
5. Wilson DR, Thiel G, Arce ML, Oken DE. Glycerol induced hemoglobinuric acute renal failure in the rat. 3. Micropuncture study of the effects of mannitol and isotonic saline on individual nephron function. Nephron 1967; 4:337-355.
6. Abul-Ezz SR, Walker PD, Shah SV. Role of glutathione in an animal model of myoglobinuric acute renal failure. Proc Natl Acad Sci U S A 1991; 88:9833-9837.
7. Zager RA. Marked protection against acute renal and hepatic injury after nitrited myoglobin+tin protoporphyrin administration. Transl Res 2015; 166:485-501.
8. Lin W, Wu X, Wen J, Fei Y, Wu J, Li X, et al. Nicotinamide retains Klotho expression and ameliorates rhabdomyolysis-induced acute kidney injury. Nutrition 2021; 91-92:111376.
9. Ni W, Zhang Y, Yin Z. The protective mechanism of Klotho gene-modified bone marrow mesenchymal stem cells on acute kidney injury induced by rhabdomyolysis. Regen Ther 2021; 18:255-267.
10. Kelly KJ, Plotkin Z, Dagher PC. Guanosine supplementation reduces apoptosis and protects renal function in the setting of ischemic injury. J Clin Invest 2001; 108:1291-1298.
11. Haynes WM. CRC Handbook of Chemistry and Physics. 97th ed. Boca Raton: CRC Press; 2016.
12. Sauriyal DS, Jaggi AS, Singh N, Muthuraman A. Investigating the role of endogenous opioids and KATP channels in glycerol-induced acute renal failure. Fundam Clin Pharmacol 2012; 26:347-355.
13. Peterson GL. Determination of total protein. Methods Enzymol 1983; 91:95-119.
14. Spanu S, van Roeyen CRC, Denecke B, Floege J, Mühlfeld AS. Urinary exosomes: a novel means to non-invasively assess changes in renal gene and protein expression. PloS One 2014; 9:e109631-e109631.
15. El-gendy F, A. H, A. S, Al Husseni N. Glutathione peroxidase-1 and klotho gene expression in acute renal failure rats and its association with creatinine and urea levels. Benha Vet Med J 2015; 29:97-104.
16. Kim S, Kim SY, Pribis JP, Lotze M, Mollen KP, Shapiro R, et al. Signaling of high mobility group box 1 (HMGB1) through toll-like receptor 4 in macrophages requires CD14. Mol Med 2013; 19:88-98.
17. Kafa IM, Uysal M, Bakirci S, Ayberk Kurt M. Sepsis induces apoptotic cell death in different regions of the brain in a rat model of sepsis. Acta Neurobiol Exp (Wars) 2010; 70:246-260.
18. Musso CG, Terrasa S, Ciocchini M, Gonzalez-Torres H, Aroca-Martinez G. Looking for a better definition and diagnostic strategy for acute kidney injury: a new proposal. Arch Argent Pediatr 2019; 117:4-5.
19. Singh AP, Junemann A, Muthuraman A, Jaggi AS, Singh N, Grover K, et al. Animal models of acute renal failure. Pharmacol Rep 2012; 64:31-44.
20. Wu J, Pan X, Fu H, Zheng Y, Dai Y, Yin Y, et al. Effect of curcumin on glycerol-induced acute kidney injury in rats. Sci Rep 2017; 7:10114-10114.
21. Boonla O, Kukongviriyapan U, Pakdeechote P, Kukongviriyapan V, Pannangpetch P, Prachaney P, et al. Curcumin improves endothelial dysfunction and vascular remodeling in 2K-1C hypertensive rats by raising nitric oxide availability and reducing oxidative stress. Nitric Oxide 2014; 42:44-53.
22. Al Asmari AK, Al Sadoon KT, Obaid AA, Yesunayagam D, Tariq M. Protective effect of quinacrine against glycerol-induced acute kidney injury in rats. BMC Nephrology 2017; 18:41.
23. Rieger E, Rech VC, Feksa LR, Wannmacher CM. Intraperitoneal glycerol induces oxidative stress in rat kidney. Clin Exp Pharmacol Physiol 2008; 35:928-933.
24. Park C, Tanaka T, Cho E, Park J, Shibahara N, Yokozawa T. Glycerol-induced renal damage improved by 7-O-Galloyl-D-sedoheptulose treatment through attenuating oxidative stress. Biol Pharm Bull 2012; 35:34-41.
25. Gudkov SV, Shtarkman IN, Smirnova VS, Chernikov AV, Bruskov VI. Guanosine and inosine display antioxidant activity, protect DNA in vitro from oxidative damage induced by reactive oxygen species, and serve as radioprotectors in mice. Radiat Res 2006; 165:538-545.
26. Roos DH, Puntel RL, Santos MM, Souza DO, Farina M, Nogueira CW, et al. Guanosine and synthetic organoselenium compounds modulate methylmercury-induced oxidative stress in rat brain cortical slices: involvement of oxidative stress and glutamatergic system. Toxicol In Vitro 2009; 23:302-307.
27. Coll E, Botey A, Alvarez L, Poch E, Quintó L, Saurina A, et al. Serum cystatin C as a new marker for noninvasive estimation of glomerular filtration rate and as a marker for early renal impairment. Am J Kidney Dis 2000; 36:29-34.
28. Nishio C, Yoshida K, Nishiyama K, Hatanaka H, Yamada M. Involvement of cystatin C in oxidative stress-induced apoptosis of cultured rat CNS neurons. Brain Res 2000; 873:252-262.
29. Yalcin S, Ulas T, Eren M, Harun A, Camuzcuoglu A, Kucuk A, et al. Relationship between oxidative stress parameters and cystatin c levels in patients with severe preeclampsia. Medicina (Kaunas, Lithuania) 2013; 49:118-123.
30. Hu MC, Shi M, Zhang J, Quinones H, Kuro-o M, Moe OW. Klotho deficiency is an early biomarker of renal ischemia-reperfusion injury and its replacement is protective. Kidney Int 2010; 78:1240-1251.
31. Hu MC, Shi M, Gillings N, Flores B, Takahashi M, Kuro OM, et al. Recombinant α-Klotho may be prophylactic and therapeutic for acute to chronic kidney disease progression and uremic cardiomyopathy. Kidney Int 2017; 91:1104-1114.
32. Hu MC, Kuro-o M, Moe OW. Klotho and chronic kidney disease. Contrib Nephrol 2013; 180:47-63.
33. Hu MC, Shi M, Zhang J, Quiñones H, Griffith C, Kuro-o M, et al. Klotho deficiency causes vascular calcification in chronic kidney disease. J Am Soc Nephrol 2011; 22:124-136.
34. Ni W, Zhang Y, Yin Z. The protective mechanism of Klotho gene-modified bone marrow mesenchymal stem cells on acute kidney injury induced by rhabdomyolysis. Regen Ther 2021; 18:255-267.
35. Oh HJ, Oh H, Nam BY, You JS, Ryu D-R, Kang S-W, et al. The protective effect of klotho against contrast-associated acute kidney injury via the antioxidative effect. Am J Physiol Renal Physiol 2019; 317:F881-F889.
36. Sharifian R, Okamura DM, Denisenko O, Zager RA, Johnson A, Gharib SA, et al. Distinct patterns of transcriptional and epigenetic alterations characterize acute and chronic kidney injury. Sci Rep 2018; 8:17870.
37. Yang J, Matsukawa N, Rakugi H, Imai M, Kida I, Nagai M, et al. Up-regulation of cAMP is a new functional signal pathway of Klotho in endothelial cells. Biochem Biophys Res Commun 2003; 301:424-429.
38. Guo Y, Zhuang X, Huang Z, Zou J, Yang D, Hu X, et al. Klotho protects the heart from hyperglycemia-induced injury by inactivating ROS and NF-kappaB-mediated inflammation both in vitro and in vivo. Biochim Biophys Acta Mol Basis Dis 2018; 1864:238-251.