Dendropanoxide Attenuates High Glucose-induced Oxidative Damage in NRK-52E Cells via AKT/mTOR Signaling Pathway

Abstract

Hyperglycemia is a potent risk factor for the development and progression of diabetes-induced nephropathy. Dendropanoxide (DPx) is a natural compound isolated from Dendropanax morbifera (Araliaceae) that exerts various biological effects. However, the role of DPx in hyperglycemia-induced renal tubular cell injury remains unclear. The present study explored the protective mechanism of DPx on high glucose (HG)-induced cytotoxicity in kidney tubular epithelial NRK-52E cells. The cells were cultured with normal glucose (5.6 mM), HG (30 mM), HG + metformin (10 µM), or HG + DPx (10 µM) for 48 h, and cell cycle and apoptosis were analyzed. Malondialdehyde (MDA), advanced glycation end products (AGEs), and reactive oxygen species (ROS) were measured. Protein-based nephrotoxicity biomarkers were measured in both the culture media and cell lysates. MDA and AGEs were significantly increased in NRK-52E cells cultured with HG, and these levels were markedly reduced by pretreatment with DPx or metformin. DPx significantly reduced the levels of kidney injury molecule-1 (KIM-1), pyruvate kinase M2 (PKM2), selenium-binding protein 1 (SBP1), or neutrophil gelatinase-associated lipocalin (NGAL) in NRK-52E cells cultured under HG conditions. Furthermore, treatment with DPx significantly increased antioxidant enzyme activity. DPx protects against HG-induced renal tubular cell damage, which may be mediated by its ability to inhibit oxidative stress through the protein kinase B/mammalian target of the rapamycin (AKT/mTOR) signaling pathway. These findings suggest that DPx can be used as a new drug for the treatment of high glucose-induced diabetic nephropathy.

Key words

Dendropanax morbifera (Araliaceae) - hyperglycemia - dendropanoxide - high glucose - advanced glycation end-product - reactive oxygen species

Publication History

Received: 18 August 2023

Accepted after revision: 30 November 2023

Accepted Manuscript online:
01 December 2023

Article published online:
16 January 2024

• References

• 1 Lin X, Xu Y, Pan X, Xu J, Ding Y, Sun X, Song X, Ren Y, Shan PF. Global, regional, and national burden and trend of diabetes in 195 countries and territories: an analysis from 1990 to 2025. Sci Rep 2020; 10: 14790
• 2 Alicic RZ, Rooney MT, Tuttle KR. Diabetic kidney disease: Challenges, progress, and possibilities. Clin J Am Soc Nephrol 2017; 12: 2032-2045
• 3 Reutens AT. Epidemiology of diabetic kidney disease. Med Clin North Am 2013; 97: 1-18
• 4 Alsaad KO, Herzenberg AM. Distinguishing diabetic nephropathy from other causes of glomerulosclerosis: An update. J Clin Pathol 2007; 60: 18-26
• 5 Giacco F, Brownlee M. Oxidative stress and diabetic complications. Circ Res 2010; 107: 1058-1070
• 6 Fiorentino T, Prioletta A, Zuo P, Folli F. Hyperglycemia-induced oxidative stress and its role in diabetes mellitus related cardiovascular diseases. Curr Pharma Des 2013; 19: 5695-5703
• 7 Popov D. Endothelial cell dysfunction in hyperglycemia: Phenotypic change, intracellular signaling modification, ultrastructural alteration, and potential clinical outcomes. Int J Diabetes Mellitus 2010; 2: 189-195
• 8 Brownlee M. The pathobiology of diabetic complications: A unifying mechanism. Diabetes 2005; 54: 1615-1625
• 9 Forbes JM, Coughlan MT, Cooper ME. Oxidative stress as a major culprit in kidney disease in diabetes. Diabetes 2008; 57: 1446-1454
• 10 Ratliff BB, Abdulmahdi W, Pawar R, Wolin MS. Oxidant mechanisms in renal injury and disease. Antioxid Redox Signal 2016; 25: 119-146
• 11 Czajka A, Malik AN. Hyperglycemia induced damage to mitochondrial respiration in renal mesangial and tubular cells: Implications for diabetic nephropathy. Redox Biol 2016; 10: 100-107
• 12 Hou S, Zheng F, Li Y, Gao L, Zhang J. The protective effect of glycyrrhizic acid on renal tubular epithelial cell injury induced by high glucose. Int J Mol Sci 2014; 15: 15026-15043
• 13 Slyne J, Slattery C, McMorrow T, Ryan MP. New developments concerning the proximal tubule in diabetic nephropathy: In vitro models and mechanisms. Nephrol Dial Transplant 2015; 30: 60-67
• 14 Vallon V, Thomson SC. Renal function in diabetic disease models: The tubular system in the pathophysiology of the diabetic kidney. Annu Rev Physiol 2012; 74: 351-375
• 15 Wada J, Makino H. Inflammation and the pathogenesis of diabetic nephropathy. Clin Sci 2013; 124: 139-152
• 16 Lu Q, Wang WW, Zhang MZ, Ma ZX, Qiu XR, Shen M, Yin XX. ROS induces epithelial-mesenchymal transition via the TGF-β1/PI3K/AKT/mTOR pathway in diabetic nephropathy. Exp Ther Med 2019; 17: 835-846
• 17 Kume S, Koya D, Uzu T, Maegawa H. Role of nutrient-sensing signals in the pathogenesis of diabetic nephropathy. Biomed Res Int 2014; 2014: 315494
• 18 Lieberthal W, Levine JS. The role of the mammalian target of rapamycin (mTOR) in renal disease. J Am Soc Nephrol 2009; 20: 2493-2502
• 19 Akram M, Kim KA, Kim ES, Syed AS, Kim CY, Lee JS, Bae ON. Potent anti-inflammatory and analgesic actions of the chloroform extract of dendropanax morbifera mediated by the Nrf2/HO-1 pathway. Biol Pharm Bull 2016; 39: 728-736
• 20 Lee D, Kim JK, Han Y, Park KI. Antihyperuricemic effect of Dendropanax morbifera leaf extract in rodent models. Evid Based Complement Alternat Med 2021; 2021: 3732317
• 21 Kim M, Park YJ, Lim HS, Lee HH, Kim TH, Lee B. The clinical effects of Dendropanax Morbifera on postmenopausal symptoms: Review article. J Menopausal Med 2017; 23: 146-155
• 22 Sachan R, Kundu A, Dey P, Son JY, Kim KS, Lee DE, Kim HR, Park JH, Lee SH, Kim JH, Cao S, Lee BM, Kwak JH, Kim HS. Dendropanax morbifera protects against renal fibrosis in streptozotocin-induced diabetic rats. Antioxidants 2020; 9: 84-107
• 23 Moon HI. Antidiabetic effects of dendropanoxide from leaves of Dendropanax morbifera Leveille in normal and streptozotocin-induced diabetic rats. Human Exp Toxicol 2011; 30: 870-875
• 24 Sachan R, Kundu A, Dey P, Son JY, Kim KS, Lee DE, Kim HR, Park JH, Lee SH, Kim JH, Cao S, Lee BM, Kwak JH, Kim HS. Dendropanax morbifera protects against renal fibrosis in streptozotocin-induced diabetic rats. Antioxidants 2020; 9: 1-20
• 25 Park YJ, Kim KS, Park JH, Lee SH, Kim HR, Lee SH, Choi HB, Cao S, Kumar V, Kwak JH, Kim HS. Protective effects of dendropanoxide isolated from Dendropanax morbifera against cisplatin-induced acute kidney injury via the AMPK/mTOR signaling pathway. Food Chem Toxicol 2020; 145: 111605
• 26 Yamagishi S, Ueda S, Matsui T, Nakamura K, Okuda S. Role of advanced glycation end products (AGEs) and oxidative stress in diabetic retinopathy. Curr Pharm Des 2008; 14: 962-968
• 27 Daenen K, Andries A, Mekahli D, Van Schepdael A, Jouret F, Bammens B. Oxidative stress in chronic kidney disease. Pediatric Nephrol 2019; 34: 975-991
• 28 Piwkowska A, Rogacka D, Audzeyenka I, Jankowski M, Angielski S. High glucose concentration affects the oxidant-antioxidant balance in cultured mouse podocytes. J Cell Biochem 2011; 112: 1661-1672
• 29 Chen SJ, Lv LL, Liu BC, Tang RN. Crosstalk between tubular epithelial cells and glomerular endothelial cells in diabetic kidney disease. Cell Prolif 2020; 53: e12763
• 30 Fiseha T. Urinary biomarkers for early diabetic nephropathy in type 2 diabetic patients. Biomark Res 2015; 3: 1-7
• 31 Kim KS, Yang HY, Song H, Kang YR, Kwon JH, An JH, Son JY, Kwack SJ, Kim YM, Bae ON, Ahn MY, Lee J, Yoon S, Lee BM, Kim HS. Identification of a sensitive urinary biomarker, selenium-binding protein 1, for early detection of acute kidney injury. J Toxicol Environ Health A 2017; 80: 453-464
• 32 Ha H, Hwang IA, Park JH, Lee HB. Role of reactive oxygen species in the pathogenesis of diabetic nephropathy. Diabetes Res Clin Pract 2008; 82: S42-45
• 33 King GL, Loeken MR. Hyperglycemia-induced oxidative stress in diabetic complications. Histochem Cell Biol 2004; 122: 333-338
• 34 Allen DA, Harwood S, Varagunam M, Raftery MJ, Yaqoob MM. High glucose-induced oxidative stress causes apoptosis in proximal tubular epithelial cells and is mediated by multiple caspases. FASEB J 2003; 17: 908-910
• 35 Maritim AC, Sanders RA, Watkins JB. Diabetes, oxidative stress, and antioxidants: A review. J Bioch Mol Toxicol 2003; 17: 24-38
• 36 Achi NK, Ohaeri OC, Ijeh II, Eleazu C. Modulation of the lipid profile and insulin levels of streptozotocin induced diabetic rats by ethanol extract of Cnidoscolus aconitifolius leaves and some fractions: Effect on the oral glucose tolerance of normoglycemic rats. Biomed Pharmacother 2017; 86: 562-569
• 37 Khalid M, Petroianu G, Adem A. Advanced glycation end products and diabetes mellitus: Mechanisms and perspectives. Biomolecules 2022; 12: 542
• 38 Wei Y, Gao J, Qin L, Xu Y, Shi H, Qu L, Liu Y, Xu T, Liu T. Curcumin suppresses AGEs induced apoptosis in tubular epithelial cells via protective autophagy. Exp Ther Med 2017; 14: 6052-6058
• 39 Donate-Correa J, Martín-Núñez E, Muros-de-Fuentes M, Mora-Fernández C, Navarro-González JF. Inflammatory cytokines in diabetic nephropathy. J Diabetes Res 2015; 2015: 948417
• 40 Meng XM, Nikolic-Paterson DJ, Lan HY. Inflammatory processes in renal fibrosis. Nat Rev Nephrol 2014; 10: 493-503
• 41 Sato M, Muragaki Y, Saika S, Roberts AB, Ooshima A. Targeted disruption of TGF-β1/Smad3 signaling protects against renal tubulointerstitial fibrosis induced by unilateral ureteral obstruction. J Clin Invest 2003; 112: 1486-1494
• 42 Wei Z, Liu HT. MAPK signal pathways in the regulation of cell proliferation in mammalian cells. Cell Res 2002; 12: 9-18
• 43 Halseth AE, Ensor NJ, White TA, Ross SA, Gulve EA. Acute and chronic treatment of ob/ob and db/db mice with AICAR decreases blood glucose concentrations. Biochem Biophy Res Commun 2002; 294: 798-805
• 44 Caton PW, Kieswich J, Yaqoob MM, Holness MJ, Sugden MC. Metformin opposes impaired AMPK and SIRT1 function and deleterious changes in core clock protein expression in white adipose tissue of genetically-obese db/db mice. Diabetes Obes Metab 2011; 13: 1097-1104
• 45 Bruggisser R, von Daeniken K, Jundt G, Schaffner W, Tullberg-Reinert H. Interference of plant extracts, phytoestrogens and antioxidants with the MTT tetrazolium assay. Planta Med 2002; 68: 445-448