Restricted accessResearch articleFirst published online 2025-5
Neuroprotective effects of SGLT2 inhibitors empagliflozin and dapagliflozin on Aβ 1–42 -induced neurotoxicity and neuroinflammation in cellular models of Alzheimer's disease
Alzheimer's disease (AD) is a chronic brain degenerative disease that leads to dementia.
Objective
The aim of the present study is to investigate the neuroprotective impact of sodium-glucose cotransporter-2 inhibitors (SGLT2i) (empagliflozin and dapagliflozin) on tau phosphorylation, oxidative stress, and neuroinflammation.
Methods
We used MTT (3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide) assay, annexin-V-FITC kit, and DCFH-DA (dichloro-dihydro-fluorescein diacetate) to respectively evaluate the effect of the SGLT2i (empagliflozin and dapagliflozin) on amyloid-β (Aβ)1–42-induced neuronal death, apoptosis, and oxidative stress. The expression of NLRP3-inflammasome, phospho-Tau181, glycogen synthase kinase-3 beta (GSK-3β), cyclin-dependent kinase 5 (CdK5), and histone deacetylase 6 (HDAC6), was quantified by flow cytometry. Drug distribution in the mice's brains was assessed by liquid chromatography-mass spectrometry (LC-MS).
Results
Aβ1–42 significantly reduced cell viability and increased apoptosis, which was reversed by using gliflozins. SGLT2i significantly reduced Aβ1–42-induced reactive oxygen species generation, downregulated NLRP3-inflammasome, and diminished tau pathology. Mechanistically, the last effect involved the modulation of GSK-3β and CdK5 protein expression. However, the tested treatments did not modify the Aβ1–42-stimulating effect of HDAC6. Gliflozins are substrates of drug transporters ATP-binding cassette sub-family B member 1 and/or ATP binding cassette subfamily G member 2 (ABCB1 and ABCG2), and Elacridar significantly enhances their brain distribution.
Conclusions
SGLT2i empagliflozin and dapagliflozin exhibited neuroprotective actions against human Aβ1–42-induced neurotoxicity.
PrinceMComas-HerreraAKnappM, et al.World Alzheimer report 2016. Improving healthcare for people living with dementia: Coverage, quality and costs now and in the future. London: Alzheimer’s Disease International, 2016.
2.
ZhouQLiSLiM, et al.Human tau accumulation promotes glycogen synthase kinase-3β acetylation and thus upregulates the kinase: a vicious cycle in Alzheimer neurodegeneration. eBioMedicine2022; 78: 103970.
3.
OchalekAMihalikBAvciHX, et al.Neurons derived from sporadic Alzheimer’s disease iPSCs reveal elevated tau hyperphosphorylation, increased amyloid levels, and GSK3B activation. Alzheimers Res Ther2017; 9: 90.
4.
DaRocha-SoutoBComaMPérez-NievasBG, et al.Activation of glycogen synthase kinase-3 beta mediates β-amyloid induced neuritic damage in Alzheimer’s disease. Neurobiol Dis2012; 45: 425–437.
5.
CrewsLPatrickCAdameA, et al.Modulation of aberrant CDK5 signaling rescues impaired neurogenesis in models of Alzheimer’s disease. Cell Death Dis2011; 2: e120.
6.
Abdel-latifRGRifaaiRAAminEF. Empagliflozin alleviates neuronal apoptosis induced by cerebral ischemia/reperfusion injury through HIF-1α/VEGF signaling pathway. Arch Pharm Res2020; 43: 514–525.
7.
WangYSanta-CruzKDecarliC, et al.NAD(P)H:quinone oxidoreductase activity is increased in hippocampal pyramidal neurons of patients with Alzheimer’s disease. Neurobiol Aging2000; 21: 525–531.
8.
Wium-AndersenIKOslerMJørgensenMB, et al.Antidiabetic medication and risk of dementia in patients with type 2 diabetes: a nested case-control study. Eur J Endocrinol2019; 181: 499–507.
9.
TangHShaoHShaabanCE, et al.Newer glucose-lowering drugs and risk of dementia: a systematic review and meta-analysis of observational studies. J Am Geriatr Soc2023; 71: 2096–2106.
10.
SiaoWZLinTKHuangJY, et al.The association between sodium-glucose cotransporter 2 inhibitors and incident dementia: a nationwide population-based longitudinal cohort study. Diabetes Vasc Dis Res2022; 19: 14791641221098168.
11.
BeelerNRiedererBMWaeberG, et al.Role of the JNK-interacting protein 1/islet brain 1 in cell degeneration in Alzheimer disease and diabetes. Brain Res Bull2009; 80: 274–281.
12.
MiklossyJQingHRadenovicA, et al.Beta amyloid and hyperphosphorylated tau deposits in the pancreas in type 2 diabetes. Neurobiol Aging2010; 31: 1503–1515.
13.
PeilaRRodriguezBLLaunerLJ. Type 2 diabetes, APOE gene, and the risk for dementia and related pathologies: the Honolulu-Asia aging study. Diabetes2002; 51: 1256–1262.
14.
BaltiraCAronicaEElmquistWF, et al.The impact of ATP-binding cassette transporters in the diseased brain: context matters. Cell Rep Med2024; 5: 101609.
15.
RobeyRWPluchinoKMHallMD, et al.Revisiting the role of efflux pumps in multidrug-resistant cancer. Nat Rev Cancer2018; 18: 452–464.
16.
HeimkeMLenzFRickertU, et al.Anti-inflammatory properties of the SGLT2 inhibitor empagliflozin in activated primary microglia. Cells2022; 11: 3107.
17.
GuglielmottoMMonteleoneDPirasA, et al.Aβ1–42 monomers or oligomers have different effects on autophagy and apoptosis. Autophagy2014; 10: 1827–1843.
18.
TamagnoEBardiniPGuglielmottoM, et al.The various aggregation states of β-amyloid 1–42 mediate different effects on oxidative stress, neurodegeneration, and BACE-1 expression. Free Radic Biol Med2006; 41: 202–212.
19.
DenizotFLangR. Rapid colorimetric assay for cell growth and survival. Modifications to the tetrazolium dye procedure giving improved sensitivity and reliability. J Immunol Methods1986; 89: 271–277.
20.
HyeoncheolKXiangX. Detection of total reactive oxygen species in adherent cells by 2′,7′-dichlorodihydrofluorescein diacetate staining. J Vis Exp2020; (160): 10.3791/60682.
21.
JóźwikALandowskiJBidzanL, et al.Beta-amyloid peptides enhance the proliferative response of activated CD4 +CD28 + lymphocytes from Alzheimer disease patients and from healthy elderly. PLoS One2012; 7: e33276.
22.
CabinioMSaresellaMPianconeF, et al.Association between hippocampal shape, neuroinflammation, and cognitive decline in Alzheimer’s disease. J Alzheimers Dis2018; 66: 1131–1144.
23.
LeccaDJungYJScerbaMT, et al.Role of chronic neuroinflammation in neuroplasticity and cognitive function: a hypothesis. Alzheimers Dement2022; 18: 2327–2340.
24.
SzczepanikAMRampeDRingheimGE. Amyloid-β peptide fragments p3 and p4 induce pro-inflammatory cytokine and chemokine production in vitro and in vivo. J Neurochem2001; 77: 304–317.
25.
United Nations System Standing Committee on Nutrition. Advancing equity, equality and non-discrimination in food systems: pathways to reform. Sixth Rep World Nutr Situat Geneva2015; 479: 132.
26.
ZhengWHBastianettoSMennickenF, et al.Amyloid β peptide induces tau phosphorylation and loss of cholinergic neurons in rat primary septal cultures. Neuroscience2002; 115: 201–211.
27.
TerwelDMuyllaertDDewachterI, et al.Amyloid activates GSK-3β to aggravate neuronal tauopathy in bigenic mice. Am J Pathol2008; 172: 786–798.
28.
HooperCKillickRLovestoneS. The GSK3 hypothesis of Alzheimer’s disease. J Neurochem2008; 104: 1433–1439.
29.
TownTZoltonJShaffnerR, et al.P35/Cdk5 pathway mediates soluble amyloid-β peptide-induced tau phosphorylation in vitro. J Neurosci Res2002; 69: 362–372.
30.
LiuPZhangZWangJ, et al.Empagliflozin protects diabetic pancreatic tissue from damage by inhibiting the activation of the NLRP3/caspase-1/GSDMD pathway in pancreatic β cells: in vitro and in vivo studies. Bioengineered2021; 12: 9356–9366.
WangCCLiYQianXQ, et al.Empagliflozin alleviates myocardial I/R injury and cardiomyocyte apoptosis via inhibiting ER stress-induced autophagy and the PERK/ATF4/Beclin1 pathway. J Drug Target2022; 30: 858–872.
33.
De BruinMMiyakeKLitmanT, et al.Reversal of resistance by GF120918 in cell lines expressing the ABC half-transporter, MXR. Cancer Lett1999; 146: 117–126.
34.
Ebrahimi-FakhariDWahlsterLBartzF, et al.Reduction of TMEM97 increases NPC1 protein levels and restores cholesterol trafficking in Niemann-pick type C1 disease cells. Hum Mol Genet2016; 25: 3588–3599.
35.
TziorasMMcGeachanRIDurrantCS, et al.Synaptic degeneration in Alzheimer disease. Nat Rev Neurol2023; 19: 19–38.
36.
AlonASchmidtHRWoodMD, et al.Identification of the gene that codes for the σ2 receptor. Proc Natl Acad Sci U S A2017; 114: 7160–7165.
37.
FunatoHYoshimuraMKusuiK, et al.Quantitation of amyloid β-protein (Aβ) in the cortex during aging and in Alzheimer’s disease. Am J Pathol1998; 152: 1633–1640.
38.
UthmanLHomayrAJuniRP, et al.Empagliflozin and dapagliflozin reduce ROS generation and restore no bioavailability in tumor necrosis factor α-stimulated human coronary arterial endothelial cells. Cell Physiol Biochem2019; 53: 865–886.
39.
UthmanLLiXBaartscheerA, et al.Empagliflozin reduces oxidative stress through inhibition of the novel inflammation/NHE/[Na+]c/ROS-pathway in human endothelial cells. Biomed Pharmacother2022; 146: 112515.
40.
CampeauMALeaskRL. Empagliflozin mitigates endothelial inflammation and attenuates endoplasmic reticulum stress signaling caused by sustained glycocalyx disruption. Sci Rep2022; 12: 12681.
41.
IannantuoniFde MarañonAMDiaz-MoralesN, et al.The SGLT2 inhibitor empagliflozin ameliorates the inflammatory profile in type 2 diabetic patients and promotes an antioxidant response in leukocytes. J Clin Med2019; 8: 1814.
42.
ArabHHEidAHAlsufyaniSE, et al.Targeting autophagy, apoptosis, and oxidative perturbations with dapagliflozin mitigates cadmium-induced cognitive dysfunction in rats. Biomedicines2023; 11: 3000.
43.
SammanWASelimSMEl FayoumiHM, et al.Dapagliflozin ameliorates cognitive impairment in aluminum-chloride-induced Alzheimer’s disease via modulation of AMPK/mTOR, oxidative stress and glucose metabolism. Pharmaceuticals2023; 16: 753.
44.
DaJXuYTanY, et al.Central administration of Dapagliflozin alleviates a hypothalamic neuroinflammatory signature and changing tubular lipid metabolism in type 2 diabetic nephropathy by upregulating MCPIP1. Biomed Pharmacother2023; 168: 115840.
45.
MohammedNNTadrosMGGeorgeMY. Empagliflozin repurposing in Parkinson’s disease; modulation of oxidative stress, neuroinflammation, AMPK/SIRT-1/PGC-1α, and wnt/β-catenin pathways. Inflammopharmacology2023; 32: 777–794.
46.
KhanTKhanSAkhtarM, et al.Empagliflozin nanoparticles attenuates type2 diabetes induced cognitive impairment via oxidative stress and inflammatory pathway in high fructose diet induced hyperglycemic mice. Neurochem Int2021; 150: 105158.
47.
ArabHHSafarMMShahinNN. Targeting ROS-dependent AKT/GSK-3β/NF-κB and DJ-1/Nrf2 pathways by dapagliflozin attenuates neuronal injury and motor dysfunction in rotenone-induced Parkinson’s disease rat model. ACS Chem Neurosci2021; 12: 689–703.
48.
Sa-nguanmooPTanajakPKerdphooS, et al.SGLT2-inhibitor And DPP-4 inhibitor improve brain function via attenuating mitochondrial dysfunction, insulin resistance, inflammation, and apoptosis in HFD-induced obese rats. Toxicol Appl Pharmacol2017; 333: 43–50.
49.
SchoonenboomNSMPijnenburgYALMulderC, et al.In CSF as markers for early-onset Alzheimer disease. Neurology2004; 62: 1580–1584.
50.
ThijssenEHLa JoieRStromA, et al.Plasma phosphorylated tau 217 and phosphorylated tau 181 as biomarkers in Alzheimer’s disease and frontotemporal lobar degeneration: a retrospective diagnostic performance study. Lancet Neurol2021; 20: 739–752.
51.
BusciglioJLorenzoAYehJ, et al.β-Amyloid fibrils induce tau phosphorylation and loss of microtubule binding. Neuron1995; 14: 879–888.
52.
HernándezFGómez de BarredaEFuster-MatanzoA, et al.GSK3: a possible link between beta amyloid peptide and tau protein. Exp Neurol2010; 223: 322–325.
53.
MaitraSVincentB. Cdk5-p25 as a key element linking amyloid and tau pathologies in Alzheimer’s disease: mechanisms and possible therapeutic interventions. Life Sci2022; 308: 12098.
54.
WuCYIskanderCWangC, et al.Association of sodium–glucose cotransporter 2 inhibitors with time to dementia: a population-based cohort study. Diabetes Care2023; 46: 297–304.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
