Senolytic combination of dasatinib and quercetin protects against diabetic kidney disease by activating autophagy to alleviate podocyte dedifferentiation via the Notch pathway

Zhu,Xinwang; Zhang,Congxiao; Liu,Linlin; Xu,Li; Yao,Li

doi:10.3892/ijmm.2024.5350

Senolytic combination of dasatinib and quercetin protects against diabetic kidney disease by activating autophagy to alleviate podocyte dedifferentiation via the Notch pathway

Authors:
- Xinwang Zhu
- Congxiao Zhang
- Linlin Liu
- Li Xu
- Li Yao
View Affiliations

Affiliations: Department of Nephrology, The First Affiliated Hospital of China Medical University, Shenyang, Liaoning 110001, P.R. China, Department of Laboratory Medicine, The Second Affiliated Hospital of Guangdong Medical University, Zhanjiang, Guangdong 524003, P.R. China
Published online on: January 17, 2024 https://doi.org/10.3892/ijmm.2024.5350
Article Number: 26
Copyright: © Zhu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The senolytics dasatinib and quercetin (DQ) alleviate age‑related disorders. However, limited information is available regarding the effects of DQ on diabetic kidney disease (DKD). The present study aimed to explore the effects of DQ on DKD and its potential molecular mechanism(s). Dasatinib (5 mg/kg) and quercetin (50 mg/kg) were administered to diabetic db/db mice by gavage for 20 weeks. Body weight, urine albumin‑creatinine ratio (ACR), serum creatinine (Scr), and blood urea nitrogen (BUN) were recorded at the indicated time periods. Periodic acid‑Schiff and Masson's staining were performed to assess the histopathological changes of kidney tissues. Immunohistochemical analysis, immunofluorescence and western blotting were performed to evaluate the expression levels of extracellular matrix (ECM) proteins, autophagic and podocyte differentiation‑related proteins. In addition, mouse podocytes were administered with high‑glucose, DQ and 3‑methyladenine (3‑MA), and the expression levels of autophagic and podocyte differentiation‑related proteins were measured. Moreover, following overexpression of the Notch intracellular domain (NICD), the expression levels of NICD, autophagic and podocyte differentiation‑related proteins were further assessed. DQ significantly reduced the body weight, blood glucose, ACR, Scr and BUN levels and improved the histopathological changes induced in diabetic db/db mice. In addition, DQ caused a significant downregulation of the expression levels of the ECM proteins, improved autophagy and induced an upregulation of the expression levels of podocyte differentiation‑related proteins. Administration of 3‑MA to mice significantly reduced podocyte differentiation, and overexpression of NICD could reverse the effects of DQ on autophagy and podocyte differentiation in vitro. The present study suggests that DQ protects against DKD by activation of autophagy to alleviate podocyte dedifferentiation via the Notch pathway.

View Figures

View References

1	Reutens AT: Epidemiology of diabetic kidney disease. Med Clin North Am. 97:1–18. 2013.
2	Hoogeveen EK: The epidemiology of diabetic kidney disease. Kidney Dial. 2:433–442. 2022.
3	Lv JC and Zhang LX: Prevalence and disease burden of chronic kidney disease. Adv Exp Med Biol. 1165:3–15. 2019.
4	Tuttle KR, Agarwal R, Alpers CE, Bakris GL, Brosius FC, Kolkhof P and Uribarri J: Molecular mechanisms and therapeutic targets for diabetic kidney disease. Kidney Int. 102:248–260. 2022.
5	Lozano R, Naghavi M, Foreman K, Lim S, Shibuya K, Aboyans V, Abraham J, Adair T, Aggarwal R, Ahn SY, et al: Global and regional mortality from 235 causes of death for 20 age groups in 1990 and 2010: A systematic analysis for the global burden of disease study 2010. Lancet. 380:2095–2128. 2012.
6	Yamazaki T, Mimura I, Tanaka T and Nangaku M: Treatment of diabetic kidney disease: Current and future. Diabetes Metab J. 45:11–26. 2021.
7	Zou H, Zhou B and Xu G: SGLT2 inhibitors: A novel choice for the combination therapy in diabetic kidney disease. Cardiovasc Diabetol. 16:652017.
8	Perkovic V, Jardine MJ, Neal B, Bompoint S, Heerspink HJL, Charytan DM, Edwards R, Agarwal R, Bakris G, Bull S, et al: Canagliflozin and renal outcomes in type 2 diabetes and nephropathy. N Engl J Med. 380:2295–2306. 2019.
9	Parzych KR and Klionsky DJ: An overview of autophagy: Morphology, mechanism, and regulation. Antioxid Redox Signal. 20:460–473. 2014.
10	Hartleben B, Gödel M, Meyer-Schwesinger C, Liu S, Ulrich T, Köbler S, Wiech T, Grahammer F, Arnold SJ, Lindenmeyer MT, et al: Autophagy influences glomerular disease susceptibility and maintains podocyte homeostasis in aging mice. J Clin Invest. 120:1084–1096. 2010.
11	Zhang C, Li W, Wen J and Yang Z: Autophagy is involved in mouse kidney development and podocyte differentiation regulated by Notch signalling. J Cell Mol Med. 21:1315–1328. 2017.
12	Yasuda-Yamahara M, Kume S, Tagawa A, Maegawa H and Uzu T: Emerging role of podocyte autophagy in the progression of diabetic nephropathy. Autophagy. 11:2385–2386. 2015.
13	Gonzales MM, Garbarino VR, Marques Zilli E, Petersen RC, Kirkland JL, Tchkonia T, Musi N, Seshadri S, Craft S and Orr ME: Senolytic therapy to modulate the progression of Alzheimer's disease (SToMP-AD): A pilot clinical trial. J Prev Alzheimers Dis. 9:22–29. 2022.
14	Novais EJ, Tran VA, Johnston SN, Darris KR, Roupas AJ, Sessions GA, Shapiro IM, Diekman BO and Risbud MV: Long-term treatment with senolytic drugs dasatinib and quercetin ameliorates age-dependent intervertebral disc degeneration in mice. Nat Commun. 12:52132021.
15	Krzystyniak A, Wesierska M, Petrazzo G, Gadecka A, Dudkowska M, Bielak-Zmijewska A, Mosieniak G, Figiel I, Wlodarczyk J and Sikora E: Combination of dasatinib and quercetin improves cognitive abilities in aged male Wistar rats, alleviates inflammation and changes hippocampal synaptic plasticity and histone H3 methylation profile. Aging (Albany NY). 14:572–595. 2022.
16	Saccon TD, Nagpal R, Yadav H, Cavalcante MB, Nunes ADC, Schneider A, Gesing A, Hughes B, Yousefzadeh M, Tchkonia T, et al: Senolytic combination of dasatinib and quercetin alleviates intestinal senescence and inflammation and modulates the gut microbiome in aged mice. J Gerontol A Biol Sci Med Sci. 76:1895–1905. 2021.
17	Levêque D, Becker G, Bilger K and Natarajan-Amé S: Clinical pharmacokinetics and pharmacodynamics of dasatinib. Clin Pharmacokinet. 59:849–856. 2020.
18	Banerjee S, Sarkar R, Mukherjee A, Miyoshi SI, Kitahara K, Halder P, Koley H and Chawla-Sarkar M: Quercetin, a flavonoid, combats rotavirus infection by deactivating rotavirus-induced pro-survival NF-κB pathway. Front Microbiol. 13:9517162022.
19	Hickson LJ, Langhi Prata LGP, Bobart SA, Evans TK, Giorgadze N, Hashmi SK, Herrmann SM, Jensen MD, Jia Q, Jordan KL, et al: Senolytics decrease senescent cells in humans: Preliminary report from a clinical trial of dasatinib plus quercetin in individuals with diabetic kidney disease. EBioMedicine. 47:446–456. 2019.
20	Palmer AK, Xu M, Zhu Y, Pirtskhalava T, Weivoda MM, Hachfeld CM, Prata LG, van Dijk TH, Verkade E, Casaclang-Verzosa G, et al: Targeting senescent cells alleviates obesity-induced metabolic dysfunction. Aging Cell. 18:e129502019.
21	Kumari R, Bettermann K, Willing L, Sinha K and Simpson IA: The role of neutrophils in mediating stroke injury in the diabetic db/db mouse brain following hypoxia-ischemia. Neurochem Int. 139:1047902020.
22	You YK, Huang XR, Chen HY, Lyu XF, Liu HF and Lan HY: C-reactive protein promotes diabetic kidney disease in db/db mice via the CD32b-Smad3-mTOR signaling pathway. Sci Rep. 6:267402016.
23	You YK, Wu WF, Huang XR, Li HD, Ren YP, Zeng JC, Chen H and Lan HY: Deletion of Smad3 protects against C-reactive protein-induced renal fibrosis and inflammation in obstructive nephropathy. Int J Biol Sci. 17:3911–3922. 2021.
24	Xu L, Fan Q, Wang X, Li L, Lu X, Yue Y, Cao X, Liu J, Zhao X and Wang L: Ursolic acid improves podocyte injury caused by high glucose. Nephrol Dial Transplant. 32:1285–1293. 2017.
25	Dungan CM, Murach KA, Zdunek CJ, Tang ZJ, Nolt GL, Brightwell CR, Hettinger Z, Englund DA, Liu Z, Fry CS, et al: Deletion of SA β-Gal+ cells using senolytics improves muscle regeneration in old mice. Aging Cell. 21:e135282022.
26	Kong ZL, Che K, Hu JX, Chen Y, Wang YY, Wang X, Lü WS, Wang YG and Chi JW: Orientin protects podocytes from high glucose induced apoptosis through mitophagy. Chem Biodivers. 17:e19006472020.
27	Zhu X, Zhang C, Shi M, Li H, Jiang X and Wang L: IL-6/STAT3-mediated autophagy participates in the development of age-related glomerulosclerosis. J Biochem Mol Toxicol. 35:e226982021.
28	Jha V, Garcia-Garcia G, Iseki K, Li Z, Naicker S, Plattner B, Saran R, Wang AY and Yang CW: Chronic kidney disease: global dimension and perspectives. Lancet. 382:260–272. 2013.
29	Huang Y, Wang B, Hassounah F, Price SR, Klein J, Mohamed TMA, Wang Y, Park J, Cai H, Zhang X and Wang XH: The impact of senescence on muscle wasting in chronic kidney disease. J Cachexia Sarcopenia Muscle. 14:126–141. 2023.
30	Li C, Shen Y, Huang L, Liu C and Wang J: Senolytic therapy ameliorates renal fibrosis postacute kidney injury by alleviating renal senescence. FASEB J. 35:e212292021.
31	Cavalcante MB, Saccon TD, Nunes ADC, Kirkland JL, Tchkonia T, Schneider A and Masternak MM: Dasatinib plus quercetin prevents uterine age-related dysfunction and fibrosis in mice. Aging (Albany NY). 12:2711–2722. 2020.
32	Adeva-Andany MM and Carneiro-Freire N: Biochemical composition of the glomerular extracellular matrix in patients with diabetic kidney disease. World J Diabetes. 13:498–520. 2022.
33	Kolset SO, Reinholt FP and Jenssen T: Diabetic nephropathy and extracellular matrix. J Histochem Cytochem. 60:976–986. 2012.
34	Canney AL, Cohen RV, Elliott JA, M Aboud C, Martin WP, Docherty NG and le Roux CW: Improvements in diabetic albuminuria and podocyte differentiation following Roux-en-Y gastric bypass surgery. Diab Vasc Dis Res. 17:14791641198790392020.
35	Weil EJ, Lemley KV, Mason CC, Yee B, Jones LI, Blouch K, Lovato T, Richardson M, Myers BD and Nelson RG: Podocyte detachment and reduced glomerular capillary endothelial fenestration promote kidney disease in type 2 diabetic nephropathy. Kidney Int. 82:1010–1017. 2012.
36	Nagata M: Podocyte injury and its consequences. Kidney Int. 89:1221–1230. 2016.
37	Mundel P, Heid HW, Mundel TM, Krüger M, Reiser J and Kriz W: Synaptopodin: An actin-associated protein in telencephalic dendrites and renal podocytes. J Cell Biol. 139:193–204. 1997.
38	Harvey SJ, Jarad G, Cunningham J, Goldberg S, Schermer B, Harfe BD, McManus MT, Benzing T and Miner JH: Podocyte-specific deletion of dicer alters cytoskeletal dynamics and causes glomerular disease. J Am Soc Nephrol. 19:2150–2158. 2008.
39	Li X, Chuang PY, D'Agati VD, Dai Y, Yacoub R, Fu J, Xu J, Taku O, Premsrirut PK, Holzman LB and He JC: Nephrin preserves podocyte viability and glomerular structure and function in adult kidneys. J Am Soc Nephrol. 26:2361–2377. 2015.
40	Nishibori Y, Liu L, Hosoyamada M, Endou H, Kudo A, Takenaka H, Higashihara E, Bessho F, Takahashi S, Kershaw D, et al: Disease-causing missense mutations in NPHS2 gene alter normal nephrin trafficking to the plasma membrane. Kidney Int. 66:1755–1765. 2004.
41	ElShaarawy A, Behairy MA, Bawady SA, Abdelsattar HA and Shadad E: Urinary podocin level as a predictor of diabetic kidney disease. J Nephropathol. 8:e262019.
42	Baisantry A, Bhayana S, Wrede C, Hegermann J, Haller H, Melk A and Schmitt R: The impact of autophagy on the development of senescence in primary tubular epithelial cells. Cell Cycle. 15:2973–2979. 2016.
43	Gewirtz DA: Autophagy and senescence: A partnership in search of definition. Autophagy. 9:808–812. 2013.
44	Gonzalez CD, Carro Negueruela MP, Nicora Santamarina C, Resnik R and Vaccaro MI: Autophagy dysregulation in diabetic kidney disease: From pathophysiology to pharmacological interventions. Cells. 10:24972021.
45	Yang D, Livingston MJ, Liu Z, Dong G, Zhang M, Chen JK and Dong Z: Autophagy in diabetic kidney disease: Regulation, pathological role and therapeutic potential. Cell Mol Life Sci. 75:669–688. 2018.
46	Huber TB, Edelstein CL, Hartleben B, Inoki K, Jiang M, Koya D, Kume S, Lieberthal W, Pallet N, Quiroga A, et al: Emerging role of autophagy in kidney function, diseases and aging. Autophagy. 8:1009–1031. 2012.
47	Barbosa Júnior Ade A, Zhou H, Hültenschmidt D, Totovic V, Jurilj N and Pfeifer U: Inhibition of cellular autophagy in proximal tubular cells of the kidney in streptozotocin-diabetic and uninephrectomized rats. Virchows Arch B Cell Pathol Incl Mol Pathol. 61:359–366. 1992.
48	Vallon V, Rose M, Gerasimova M, Satriano J, Platt KA, Koepsell H, Cunard R, Sharma K, Thomson SC and Rieg T: Knockout of Na-glucose transporter SGLT2 attenuates hyperglycemia and glomerular hyperfiltration but not kidney growth or injury in diabetes mellitus. Am J Physiol Renal Physiol. 304:F156–F167. 2013.
49	Matboli M, Eissa S, Ibrahim D, Hegazy MGA, Imam SS and Habib EK: Caffeic acid attenuates diabetic kidney disease via modulation of autophagy in a high-fat diet/streptozotocin-induced diabetic rat. Sci Rep. 7:22632017.
50	Ren H, Shao Y, Wu C, Ma X, Lv C and Wang Q: Metformin alleviates oxidative stress and enhances autophagy in diabetic kidney disease via AMPK/SIRT1-FoxO1 pathway. Mol Cell Endocrinol. 500:1106282020.
51	Zheng D, Tao M, Liang X, Li Y, Jin J and He Q: p66Shc regulates podocyte autophagy in high glucose environment through the Notch-PTEN-PI3K/Akt/mTOR pathway. Histol Histopathol. 35:405–415. 2020.
52	Yoshida G, Kawabata T, Takamatsu H, Saita S, Nakamura S, Nishikawa K, Fujiwara M, Enokidani Y, Yamamuro T, Tabata K, et al: Degradation of the NOTCH intracellular domain by elevated autophagy in osteoblasts promotes osteoblast differentiation and alleviates osteoporosis. Autophagy. 18:2323–2332. 2022.
53	Niranjan T, Bielesz B, Gruenwald A, Ponda MP, Kopp JB, Thomas DB and Susztak K: The Notch pathway in podocytes plays a role in the development of glomerular disease. Nat Med. 14:290–298. 2008.
54	Walsh DW, Roxburgh SA, McGettigan P, Berthier CC, Higgins DG, Kretzler M, Cohen CD, Mezzano S, Brazil DP and Martin F: Co-regulation of Gremlin and Notch signalling in diabetic nephropathy. Biochim Biophys Acta. 1782:10–21. 2008.
55	Waters AM, Wu MYJ, Onay T, Scutaru J, Liu J, Lobe CG, Quaggin SE and Piscione TD: Ectopic notch activation in developing podocytes causes glomerulosclerosis. J Am Soc Nephrol. 19:1139–1157. 2008.
56	Wang C, Kang Y, Liu P, Liu W, Chen W, Hayashi T, Mizuno K, Hattori S, Fujisaki H and Ikejima T: Combined use of dasatinib and quercetin alleviates overtraining-induced deficits in learning and memory through eliminating senescent cells and reducing apoptotic cells in rat hippocampus. Behav Brain Res. 440:1142602023.
57	Maiuri MC, Zalckvar E, Kimchi A and Kroemer G: Self-eating and self-killing: Crosstalk between autophagy and apoptosis. Nat Rev Mol Cell Biol. 8:741–752. 2007.
58	Scarlatti F, Granata R, Meijer AJ and Codogno P: Does autophagy have a license to kill mammalian cells? Cell Death Differ. 16:12–20. 2009.
59	Rubinstein AD and Kimchi A: Life in the balance-a mechanistic view of the crosstalk between autophagy and apoptosis. J Cell Sci. 125:5259–5268. 2012.